WO2024077205A2 - Moraxellaceae o-linking oligosaccharyltransferases, glycosylation fragments, and uses thereof - Google Patents

Abstract

A method of producing a glycoconjugate comprising covalently linking an oligo- or polysaccharide to an acceptor protein comprising or consisting of a TfpM-associated pilin-like protein or glycosylation fragment thereof using a TfpM oligosaccharyltransferase (OTase), and compositions thereof.

Description

MORAXELLACEAE O-LINKING OLIGOSACCHARYLTRANSFERASES, GLYCOSYLATION FRAGMENTS, AND USES THEREOF Inventors: Cory James Knoot Lloyd Sherwood Robinson Christian Michael Harding CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of U.S. Provisional Patent Application No. 63/414,185, filed October 7, 2022, which is incorporated herein by reference in its entirety. STATEMENT REGARDING FEDERALLY-SPONSORED RESEARCH AND DEVELOPMENT [0002] This invention was made with the government support under the R44AI131742 grant awarded by the National Institute for Allergy and Infectious Disease (NIAID). The Government has certain rights in the invention. REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY [0003] The contents of the electronically submitted sequence listing in ST26 format (Name VAXNEWMO_229640_ST26_SeqList.xml; Size: 194,162 bytes; and Date of Creation: October 5, 2023) filed with this application is incorporated herein by reference in its entirety. BACKGROUND Field of the Invention [0004] This disclosure is directed to the field of in vivo protein glycosylation and conjugate vaccine development. [0005] Protein glycosylation is the most common post-translational modification and is prevalent in all three domains of life. In bacteria, protein glycosylation can be mediated directly by glycosyltransferases that sequentially glycosylate acceptor proteins with individual monosaccharides or by oligosaccharyltransferases (OTases) that transfer a preassembled oligosaccharide en bloc to acceptor proteins. Bacterial oligosaccharyltransferases comprise a family of glycosyltransferases that fall into two major categories: N-linking and O-linking (Harding, C. M., and Feldman, M. F. (2019) Glycobiology 29, 519-529; Nothaft, H., and Atty. Dkt. No.64100-229640 Szymanski, C. M. (2010) Nature Reviews Microbiology 8, 765-778). N-linking oligosaccharyltransferases catalyze the covalent linkage of pre-assembled oligosaccharides to the sidechains of asparagine residues in acceptor proteins, whereas O-linking oligosaccharyltransferases typically transfer glycans to the sidechains of serine or threonine residues (Nothaft, H., and Szymanski, C. M. (2010) Nature Reviews Microbiology 8, 765-778). The amino acid motifs on substrate proteins that are recognized by oligosaccharyltransferases are termed sequons. Both N- and O-linking oligosaccharyltransferases are membrane-bound enzymes residing in the inner membrane and glycosylate proteins in the periplasmic space with glycans derived from lipid-linked precursors. The sugar substrate for these glycosylation reactions usually derives from a highly conserved pathway that synthesizes lipid-linked oligosaccharides on the inner leaflet of the cytoplasmic membrane that are ‘flipped’ to the outer leaflet in a manner analogous to the Wzy-dependent pathway for O-antigen biosynthesis (Raetz, C. R. H., and Whitfield, C. (2002) Annual Review of Biochemistry 71, 635-700). The lipid carrier molecule for these reactions is typically undecaprenol pyrophosphate (Und-PP) and oligosaccharyltransferases catalyze the transfer of Und-PP-linked glycans to acceptor proteins. Many O-linking oligosaccharyltransferases have been shown to be promiscuous with their polysaccharide substrate and are able to transfer a variety of different bacterial glycans to acceptor proteins (Faridmoayer, A., et al. (2008) Journal of Biological Chemistry 283, 34596-34604; DiGiandomenico, A., et al. (2002) Molecular Microbiology 46, 519-530). These bacterial glycans are typically composed of repeating sets of two to seven monosaccharides known as repeat units that are polymerized into polysaccharides in the periplasm prior to transfer (Harding, C. M., and Feldman, M. F. (2019) Glycobiology 29, 519-529). Aside from providing basic insights into bacterial glycosylation pathways, oligosaccharyltransferases have garnered significant interest due to their ability to transfer diverse bacterial O-antigen and capsular polysaccharides to specific residues on engineered carrier proteins in a process termed bioconjugation or protein-glycan coupling technology (Harding, C. M., and Feldman, M. F. (2019) Glycobiology 29, 519-529; Feldman, M. F., et al. (2005) Proceedings of the National Academy of Sciences of the United States of America 102, 3016; Kay, E. J., et al. (2016) Open Biology 6, 150243; Kay, E., et al. (2019) npj Vaccines 4, 16). The resulting bioconjugate glycoproteins have been leveraged as glycoconjugate vaccines for a variety of bacterial pathogens with multiple candidates currently in clinical trials (GSK 2022 Infectious Disease Pipeline (2022) Found on the world wide web at gsk.com/en-gb/research-and- development/our-pipeline/?infectious-diseases; Johnson & Johnson Infectious Diseases and Atty. Dkt. No.64100-229640 Vaccines, Global Public Health Pipeline (2022) Found on the world wide web at investor.jnj.com/pharmaceutical-pipeline-information). [0006] Among other substrates, O-linking oligosaccharyltransferases most notably catalyze the glycosylation of type IV pilin-like proteins (Schäffer, C., and Messner, P. (2017) FEMS Microbiology Reviews 41, 49-91). Type IV pili are composed primarily of protein subunits called major pilins that non-covalently interact to form the fibrous structure of the pilus shaft (Giltner Carmen, L., et al. (2012) Microbiology and Molecular Biology Reviews 76, 740-772). The roles of pilin glycosylation in the bacterial lifestyle are numerous. Pilus glycosylation has been shown to aid in bacterial phage defense by blocking the binding of phage particles to surface proteins (Harvey, H., et al. (2018) Nature Microbiology 3, 47-52). In pathogenic bacteria such as Neisseria, Pseudomonas, Burkholderia and others, pilin glycosylation has a wide variety of effects leading to increased virulence (Yakovlieva, L., et al. (2021) Frontiers in Microbiology 12) by modulating tissue adherence and invasion (Marceau, M., et al. (1998) Molecular Microbiology 27, 705-715; Willcocks, S. J., et al. (2020) Future Microbiology 15, 241-257; Nguyen, L. C., et al. (2012) Molecular Plant Pathology 13, 764-774) and by blocking the binding of complement proteins leading to immune system evasion (Tan Rommel, et al. (2015) Infection and Immunity 83, 1339- 1346). This lends importance to studies exploring the potential of bacterial glycosylation systems as novel antimicrobial and anti-virulence targets (Yakovlieva, L., et al. (2021) Frontiers in Microbiology 12). [0007] Historically, O-linking oligosaccharyltransferases have been functionally classified based on several characteristics: (i) the types of substrate proteins that the oligosaccharyltransferase glycosylates, (ii) the types of glycans that the enzymes are capable of transferring, often defined by the monosaccharide or disaccharide at the saccharide reducing end (the sugar covalently linked to Und-PP), (iii) the location of the glycosylated amino acid on substrate proteins, and (iv) the typical number of sugar monomers or oligosaccharide repeat units in the glycan that is transferred to the acceptor protein. Three major types of bacterial O-linking OTases have been previously identified and archetypal representatives from Neisseria (Faridmoayer, A., et al. (2007) Journal of Bacteriology 189, 8088-8098), Pseudomonas (Castric, P. (1995) Microbiology 141, 1247-1254; Horzempa, J., et al. (2006) Journal of Biological Chemistry 281, 1128-1136; Harvey, H., et al. (2009) Journal of Bacteriology 191, 6513-6524; Qutyan, M., et al. (2010) Journal of Bacteriology 192, 5972-5981), and Acinetobacter (Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041; Harding, C. M., et al. (2019) Nature Atty. Dkt. No.64100-229640 Communications 10, 891; Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203) have been characterized. TfpO (formerly known as PilO) was the first bacterial oligosaccharyltransferase discovered and exclusively catalyzes the glycosylation of a single protein PilA that is the major pilin subunit of type IV pili (Giltner Carmen, L., et al. (2012) Microbiology and Molecular Biology Reviews 76, 740-772). TfpO was first characterized in Pseudomonas aeruginosa strain 1244 (P. aeruginosa 1244) (Castric, P. (1995) Microbiology 141, 1247-1254; Comer Jason, E., et al. (2002) Infection and Immunity 70, 2837-2845) but, more recently, TfpO orthologs from other organisms have been characterized in medically relevant Acinetobacter species (Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041). TfpO proteins catalyze the attachment of a single O- antigen repeat unit to the extreme C-terminal serine residue on PilA (Comer Jason, E., et al. (2002) Infection and Immunity 70, 2837-2845). PglL (also referred to as PglO) proteins are another class of bacterial O-linking oligosaccharyltransferases and are considered ‘general’ oligosaccharyltransferases, catalyzing the glycosylation of multiple periplasmic and membrane- associated proteins including the major pilin subunit PilE of Neisseria type IV pili (Faridmoayer, A., et al. (2007) Journal of Bacteriology 189, 8088-8098; Vik, Å., et al. (2009) Proceedings of the National Academy of Sciences 106, 4447; Hayes, A. J., et al. (2021) Communications Biology 4, 1045). PglL orthologs naturally transfer glycans with either 2-N-acetyl sugars or the hexose galactose at the reducing end of the glycan (Faridmoayer, A., et al. (2008) Journal of Biological Chemistry 283, 34596-34604), but do not naturally transfer glycans with glucose as a reducing end sugar (Harding, C. M., et al. (2019) Nature Communications 10, 891). PglS orthologs are the third type of O-linking bacterial oligosaccharyltransferase and are the most recently discovered of the three types (Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041). PglS from Acinetobacter baylyi ADP1 (PglS_ADP1) is the most well-characterized and natively catalyzes the glycosylation of a single pilin protein called ComP that forms type IV-like pili involved in natural competence (Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041; Porstendörfer, D., et al. (2000) Journal of Bacteriology 182, 3673-3680). Notably, PglSADP1 has the broadest substrate scope of all oligosaccharyltransferases characterized, transferring virtually any glycan to ComP including those having 2-N-acetyl sugars, galactose, or glucose at the reducing end (Harding, C. M., et al. (2019) Nature Communications 10, 891). [0008] Notwithstanding differences in sequence and ostensibly protein structure, there are several notable distinctions between TfpO, PglL, and PglS proteins. First, TfpO proteins glycosylate the C-terminal serine or threonine residue of substrate proteins (Comer Jason, E., et al. Atty. Dkt. No.64100-229640 (2002) Infection and Immunity 70, 2837-2845), whereas PglL and PglS proteins glycosylate sequons that are in the interior of the acceptor protein amino acid sequence (Harding, C. M., et al. (2019) Nature Communications 10, 891; Vik, Å., et al. (2009) Proceedings of the National Academy of Sciences 106, 4447). Second, TfpO proteins are limited to transferring only short oligosaccharides to acceptor proteins, while PglL and PglS proteins, when heterologously expressed in glycoengineered E. coli systems, can transfer long-chain polysaccharides with multiple repeat units composed of possibly hundreds of monosaccharides (Faridmoayer, A., et al. (2007) Journal of Bacteriology 189, 8088-8098; Horzempa, J., et al. (2006) Journal of Biological Chemistry 281, 1128-1136; Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041; Harding, C. M., et al. (2019) Nature Communications 10, 891; Comer Jason, E., et al. (2002) Infection and Immunity 70, 2837-2845). However, the reason for this difference in glycan substrate size and reducing-end sugar preference are not known, in part due to the unavailability of protein structures of any bacterial O-linking oligosaccharyltransferases. [0009] Thus, there remains a need to identify novel members of the bacterial O-linking OTase family. Comparisons of known bacterial O-linking OTases and new representatives would facilitate studies of OTase structure-function relationships and potentially identify the determinants of OTase-glycan specificity. SUMMARY [0010] This disclosure is directed to a glycoconjugate comprising an oligo- or polysaccharide covalently linked to an acceptor protein, wherein the acceptor protein comprises or consists of a TfpM-associated pilin-like protein or glycosylation fragment thereof and the oligo- or polysaccharide is covalently linked to the pilin-like protein or glycosylation fragment thereof, and wherein the TfpM-associated pilin-like protein or glycosylation fragment thereof comprises a C- terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C- terminus serine or threonine. In certain embodiments, the acceptor protein is a fusion protein comprising the TfpM-associated pilin-like protein or glycosylation fragment thereof translationally fused to a heterologous carrier protein and the TfpM-associated pilin-like protein or glycosylation fragment thereof is the C-terminus-most sequence of the acceptor protein such that the acceptor fusion protein comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine. In certain embodiments, the glycoconjugate is immunogenic. Atty. Dkt. No.64100-229640 [0011] This disclosure is also directed to a pilin-like-protein glycosylation fragment comprising or consisting of an isolated fragment of a TfpM-associated pilin-like protein, wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is PilMo (SEQ ID NO: 57) or PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3 .1$ VY H polypeptide comprising at least comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, optionally, wherein the C- terminus threonine is substituted with serine; wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil_72-O-c (SEQ ID NO: 93), Pil_BI730 (SEQ ID NO: 94), Pil_A3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), Pil_T-3-2 (SEQ ID NO: 99), Pil_CIP102637 (SEQ ID NO: 100), and a fragment (e.g., C-terminus fragment) of any thereof and/or a variant wherein the C-terminus threonine is substituted with serine; and/or wherein the pilin-like-protein glycosylation fragment comprises or consists of the Pil_Mo pilin disulfide loop region (Pil_Mo_DSL, also referred to as Pil₂₀; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148). In certain embodiments, the pilin-like-protein glycosylation fragment consists of Pil₂₀ (SEQ ID NO: 60), Pil₁₉ (SEQ ID NO: 133), Pil₁₈ (SEQ ID NO: 134), Pil₁₇ (SEQ ID NO: 135), Pil₁₆ (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil₁₁ (SEQ ID NO: 139), Pil₁₀ (SEQ ID NO: 112), Pil₉ (SEQ ID NO: 140), Pil₈ (SEQ ID NO: 141), Pil7 (SEQ ID NO: 113), Pil6 (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine. In certain embodiments, the pilin- like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Atty. Dkt. No.64100-229640 Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine. [0012] This disclosure is also directed to a fusion protein comprising a TfpM-associated pilin- like protein or glycosylation fragment thereof translationally fused to a heterologous carrier protein, wherein the pilin-like protein or glycosylation fragment comprises a C-terminus serine or threonine residue, wherein the pilin-like protein or glycosylation fragment is the C-terminus-most sequence of the fusion protein, and wherein the fusion protein comprises a C-terminus serine or threonine residue. In certain embodiments, the fusion protein is glycosylated by an oligo- or polysaccharide comprising glucose at its reducing end covalently linked to the C-terminus serine or threonine. [0013] This disclosure is directed to a method of producing a glycoconjugate comprising covalently linking an oligo- or polysaccharide to an acceptor protein comprising or consisting of a TfpM-associated pilin-like protein or glycosylation fragment thereof using a TfpM oligosaccharyltransferase (OTase), wherein the pilin-like protein or glycosylation fragment comprises a C-terminus serine or threonine residue, the acceptor protein comprises a C-terminus serine or threonine residue, and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine residue of the acceptor protein. In certain embodiments the method is a method of in vivo conjugation of an oligo- or polysaccharide to an acceptor protein. In certain embodiments, the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpM_Mo (SEQ ID NO: 56), TfpM_DSM16617 (SEQ ID NO: 63), TfpM_ZZC3 (SEQ ID NO: 64), TfpM_TUM15069 (SEQ ID NO: 65), TfpM_AI7 (SEQ ID NO: 66), TfpM_VE- C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpM_F78 (SEQ ID NO: 71), TfpM_S71 (SEQ ID NO: 72), TfpM_ANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81). In certain embodiments, the TfpM OTase is TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), Atty. Dkt. No.64100-229640 TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), or TfpM72-O-c (SEQ ID NO: 81). [0014] This disclosure provides for an isolated nucleic acid encoding the pilin-like-protein glycosylation fragment and/or the fusion protein above and any of their embodiments described anywhere herein and also for a host cell comprising the isolated nucleic acid. [0015] This disclosure provide for a method of inducing a host immune response against a bacterial pathogen, the method comprising administering to a subject in need of the immune response an effective amount of a glycoconjugate, conjugate vaccine, fusion protein, or composition of either thereof disclosed anywhere herein. [0016] This disclosure provides for a method of preventing or treating a bacterial disease and/or infection in a subject comprising administering to a subject in need thereof a glycoconjugate, conjugate vaccine, a fusion protein, or a composition of either thereof disclosed anywhere herein. [0017] This disclosure provides for a method of producing a pneumococcal conjugate vaccine against pneumococcal infection, comprising: (a) isolating a glycoconjugate or a glycosylated fusion protein disclosed anywhere herein; and (b) combining the isolated glycoconjugate or isolated glycosylated fusion protein with an adjuvant and/or carrier. [0018] This disclosure provides for a recombinant nucleic acid construct comprising a nucleotide sequence encoding a TfpM oligosaccharyltransferase (OTase) operably linked to at least one heterologous transcriptional regulatory sequence. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof operably linked to the nucleotide sequence encoding the a TfpM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof 5’ of and operably linked to the nucleotide sequence encoding the a TfpM OTase. BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES [0019] Figure 1A, B, C, and D. Figure 1 shows the characterization of 13 TfpM orthologs from species in the Moraxellaceae family. A) Cladogram of the 20 TfpM orthologs with archetypal TfpO, PglL, and PglS from Pseudomonas, Neisseria, and Acinetobacter, respectively. Branch Atty. Dkt. No.64100-229640 confidence is indicated in red. Starred OTase/pilin pairs were cloned and tested in bioconjugation experiments. B) Diagram showing the EPA-pilin fusion protein and TfpM construct design. Colored arrows indicate genes. Gene expression was driven from an IPTG-inducible tac promoter with a lacO operator (tac1O). The rrnB T2 terminator is marked by a black hairpin structure. C) and D) Anti-EPA western blot of whole-cell E. coli extracts expressing different EPA-pilin carrier open reading frames and tfpM genes. Panel D shows the same image as panel C but at higher exposure. Whole cell extracts loaded in each lane were normalized by OD600. H286A indicates the M. osloensis TfpM site directed OTase active site mutant. “g0” indicates unglycosylated EPA-pilin and “gn” indicates CPS8-glycosylated EPA-pilin protein. Reference protein masses are marked left of the western blot in kDa. [0020] Figure 2. Figure 2 shows a phylogram of TfpM, PilO, PglL, and PglS orthologs with relative distances shown. The phylogenetic tree was generated using the phylogeny.fr server (on the world wide web at phylogeny.fr/) which uses MUSCLE, PhyML, and TreeDyn for sequence alignment, tree calculation, and image generation, respectively. [0021] Figure 3. Figure 3 Phylogram of TfpM-associated pilin-like proteins, select PilA proteins from Neisseria and Pseudomonas, and ComP from A. soli CIP 110264 with relative distances shown. The red numbers indicate branch confidence. The phylogenetic tree was generated using the phylogeny.fr server (on the world wide web at phylogeny.fr/) which uses MUSCLE, PhyML, and TreeDyn for sequence alignment, tree calculation, and image generation, respectively. [0022] Figure 4. Figure 4 shows multiple sequence alignment of select bacterial O-linking oligosaccharyltransferase. The alignment was generated using Clustal Omega with default settings found on the world wide web at ebi.ac.uk/Tools/msa/clustalo/. N_menigitidis_MC58_PglL (SEQ ID NO: 105). A_baylyi_ADP1_PglS (SEQ ID NO: 106); P_aeruginosa_1244_TfPO (SEQ ID NO: 107); M_osloensis_1202_TfpM (SEQ ID NO: 56); A_nosocomialis_M2_TfpO (SEQ ID NO: 108). [0023] Figure 5. Figure 5 shows an anti-EPA whole-cell western blot examining the glycosylation status of the EPA-PilMob+1 M\ZPVUZ HZ ^LSS HZ 9B5&BPSMob+1 7&[LYTPUHS FOY¹⁶⁷ mutants. All lanes were normalized to the same OD600. Reference protein masses are marked next to the western blot in kDa. [0024] Figure 6A and B. Figure 6 shows Targeted MS/MS analysis of the HexHexA- modified C-terminal EPA-PilMob+1 WLW[PKL ⁷⁶²FLPANCRGT⁷⁷⁰ (SEQ ID NO: 61). A) EThcD fragmentation enabled the localization of the HexHexA glycosylation event to the terminal residue Atty. Dkt. No.64100-229640 Thr⁷⁷⁰. B) HCD fragmentation enables the confirmation of the peptide sequence as well as the linkage of the disaccharide HexHexA through the Hex monosaccharide by the observation of multiple y-ions linked solely to a Hex residue. [0025] Figure 7A,B,C,D,E,F and G. Figure 7 shows TfpMMo can transfer diverse bacterial glycans to the EPA-PilMoH28 fusion protein. A) Structures of the repeat units of the five bacterial glycans tested with TfpMMo. The linkages between sugar monomers are indicated in rounded brackets. Glycan abbreviations used: CPS8, S. pneumoniae capsular polysaccharide Type 8; GBSIII, Group B Streptococcus capsular polysaccharide Type III; LT2, Salmonella enterica Group B serotype LT2 O-antigen; O16, E. coli serotype O16 O-antigen; O2a, Klebsiella pneumoniae serotype O2a O-antigen; All sugars are the pyranose form except where noted. Abbreviations used: Glc, glucose; Gal, galactose; Galf, galactofuranose; Rha, rhamnose; GlcNAc, N- acetylglucosamine; Abe, abequose; NeuNAc, N-acetylneuraminic acid, sialic acid. B)-F) Anti- glycan western blots with partially purified TfpMMo-derived bioconjugates. B) Anti-CPS8. C) Anti-O16. D) Anti-LT2. E) Anti-O2a. F) Anti-GBSIII. G) Anti-EPA. In panels B)-G), +/- labels indicate whether the samples were incubated with (+) or without (-) proteinase K prior to SDS- PAGE separation. Reference protein masses are marked next to the western blots in kDa. [0026] Figure 8A and B. Figure 8 shows TfpMMo glycosylates truncated EPA-fused pilin variants as small as three amino acids. A) Sequences of the EPA-fused PilMo fragments tested for bioconjugation with TfpMMo. Blue letters mark the C-terminal residues of EPA (i.e., EDLK; SEQ ID NO: 132). Underlined residues indicate the glycine linker placed between EPA and the pilin sequence. B) Anti-EPA western blot of whole cell extracts expressing the truncated pilin variants, CPS8, and TfpM_Mo. The calculated EPA-Pil_Mob+1 THZZ PZ 1)', R8H HUK [OH[ VM [OL [Y\UJH[LK variants ranges from 67.1 to 69.0 kDa. All lanes were normalized to the same OD₆₀₀. Reference protein masses are marked left of the western blot in kDA. “g₀” indicates the unglycosylated truncated EPA-pilins and “g_n” indicates the glycosylated EPA-pilin protein. Unglycosylated EPA- PilMob+1 Y\UZ ULHY 0. R8H' BPS20 (SEQ ID NO: 60). Pil15[A] (SEQ ID NO: 171). Pil13[A] (SEQ ID NO: 173). GGGG plus Pil_10[A] is Pil_10L[A] (SEQ ID NO: 111). Pil_10[A] (SEQ ID NO: 176). Pil_7[A] (SEQ ID NO: 179). Pil6[A] (SEQ ID NO: 180). Pil5[A] (SEQ ID NO: 181). Pil4[A] (SEQ ID NO: 182). Pil3 “RGT” (SEQ ID NO: 117). EDLK plus Pil2 (SEQ ID NO: 118). EDLKGGGG plus Pil20 (SEQ ID NO: 122). EDLK plus Pil15[A] (SEQ ID NO: 123). EDLK plus Pil13[A] (SEQ ID NO: 124). EDLK plus Pil10L[A] (SEQ ID NO: 125). EDLK plus Pil10[A] (SEQ ID NO: 126). EDLK plus Pil7[A] Atty. Dkt. No.64100-229640 (SEQ ID NO: 127). EDLK plus Pil6[A] (SEQ ID NO: 128). EDLK plus Pil5[A] (SEQ ID NO: 129). EDLK plus Pil4[A] (SEQ ID NO: 130). EDLK plus Pil3 (SEQ ID NO: 131). [0027] Figure 9. Figure 9 shows multiple sequence alignment of select pilin proteins. Accession numbers for these proteins are given in the main text. The alignment was generated using Clustal Omega with default settings found on the world wide web at ebi.ac.uk/Tools/msa/clustalo/. P_aeruginosa_1244_PilA (SEQ ID NO: 119). N_menigitidis_M2_PilA (SEQ ID NO: 120). A_junii_65_pilin (SEQ ID NO: 97). A_CIP102143_pilin (SEQ ID NO: 88). A_CIP102637_pilin (SEQ ID NO: 100). A_YZS-X1- 1_pilin (SEQ ID NO: 98). A_soli_110264_ComP (SEQ ID NO: 121). A_YH01026_pilin (SEQ ID NO: 87). M_osloensis_1202_pilin (SEQ ID NO: 57). A_junii_TUM15069_pilin (SEQ ID NO: 84). [0028] Figure 10A,B,C,D,E, and F. Figure 10 shows purified TfpMMo-derived GBSIII bioconjugate elicits a robust IgG immune response in mice. A) Western blot of purified GBSIII- 291 bioconjugate, anti-EPA channel. B) Anti-GBSIII. C) Merged image of A and B. D) Coomassie stain of purified GBSIII-291 bioconjugate. E) MS1 spectrum of intact, purified GBSIII-291 bioconjugate. The 291 protein (EPA-Pil₂₀) has a theoretical mass of 69,582.19 Da. The GBSIII- 291 bioconjugate is observed in multiple states of increasing mass separated by near 980 Da that corresponds to the calculated mass of a GBSIII glycan repeat unit. F) GBSIII- specific IgG kinetics over the course of immunization as measured by ELISA and converted to ng/mL IgG using a standard IgG curve. **P<0.01. [0029] Figure 11A,B. Figure 11 shows glycosylation of EPA constructs containing sequons from different O-linking oligosaccharyltransferase systems. A) Diagrams of the plasmid-based operons expressing EPA with PglS- or TfpM-specific sequons. Internal glycotag (“iGT”) is a 23 amino acid fragment derived from ComP₁₁₀₂₆₄ inserted between EPA Ala⁴⁸⁹ – Arg⁴⁹⁰ and/or Glu⁵⁴⁸ – Gly⁵⁴⁹. B) Anti-EPA western blot of SDB1 periplasmic extracts expressing one of the four constructs and the E. coli O16 O-antigen. Load amounts per lane were normalized to OD600. “g0” indicates unglycosylated EPA carrier protein and the singly or doubly glycosylated EPA proteins are indicated. Reference protein masses are marked left of the western blot in kDa. DETAILED DESCRIPTION [0030] To the extent necessary to provide descriptive support, the subject matter and/or text of the appended claims is incorporated herein by reference in their entirety. Atty. Dkt. No.64100-229640 [0031] It will be understood by all readers of this written description that the exemplary aspects and embodiments described and claimed herein may be suitably practiced in the absence of any recited feature, element or step that is, or is not, specifically disclosed herein. Definitions [0032] It is to be noted that the term "a" or "an" entity refers to one or more of that entity; for example, "a polysaccharide," is understood to represent one or more polysaccharides. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein. [0033] Furthermore, "and/or" where used herein is to be taken as specific disclosure of each of the specified features or components with or without the other. Thus, the term and/or" as used in a phrase such as "A and/or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and/or" as used in a phrase such as "A, B, and/or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). [0034] It is understood that wherever aspects are described herein with the language "comprising" or “comprises” otherwise analogous aspects described in terms of "consisting of," “consists of,” "consisting essentially of," and/or “consists essentially of,” and the like are also provided. [0035] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. [0036] Numeric ranges are inclusive of the numbers defining the range. Even when not explicitly identified by “and any range in between,” or the like, where a list of values is recited, e.g., 1, 2, 3, or 4, unless otherwise stated, the disclosure specifically includes any range in between the values, e.g., 1 to 3, 1 to 4, 2 to 4, etc. [0037] The headings provided herein are solely for ease of reference and are not limitations of the various aspects or aspects of the disclosure, which can be had by reference to the specification as a whole. [0038] As used herein, the term “non-naturally occurring” substance, composition, entity, and/or any combination of substances, compositions, or entities, or any grammatical variants thereof, is a conditional term that explicitly excludes, but only excludes, those forms of the substance, composition, entity, and/or any combination of substances, compositions, or entities that are well-understood by persons of ordinary skill in the art as being “naturally-occurring,” or Atty. Dkt. No.64100-229640 that are, or might be at any time, determined or interpreted by a judge or an administrative or judicial body to be, “naturally-occurring.” [0039] As used herein, the term “polypeptide” is intended to encompass a singular “polypeptide” as well as plural “polypeptides,” and refers to a molecule composed of monomers (amino acids) linearly linked by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any chain or chains of two or more amino acids, and does not refer to a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, “protein,” “amino acid chain,” or any other term used to refer to a chain or chains of two or more amino acids are included within the definition of "polypeptide,” and the term “polypeptide” can be used instead of, or interchangeably with any of these terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of the polypeptide, including without limitation glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, or modification by non-standard amino acids. A polypeptide can be derived from a natural biological source or produced by recombinant technology, but is not necessarily translated from a designated nucleic acid sequence. It can be generated in any manner, including by chemical synthesis. [0040] A “protein” as used herein can refer to a single polypeptide, i.e., a single amino acid chain as defined above, but can also refer to two or more polypeptides that are associated, e.g., by disulfide bonds, hydrogen bonds, or hydrophobic interactions, to produce a multimeric protein. [0041] By an "isolated" polypeptide or a fragment, variant, or derivative thereof is intended a polypeptide that is not in its natural milieu. No particular level of purification is required. For example, an isolated polypeptide can be removed from its native or natural environment. Recombinantly produced polypeptides and proteins expressed in host cells are considered isolated as disclosed herein, as are recombinant polypeptides that have been separated, fractionated, or partially or substantially purified by any suitable technique. [0042] As used herein, the term “non-naturally occurring” polypeptide, or any grammatical variants thereof, is a conditional term that explicitly excludes, but only excludes, those forms of the polypeptide that are well-understood by persons of ordinary skill in the art as being “naturally- occurring,” or that are, or might be at any time, determined or interpreted by a judge or an administrative or judicial body to be, “naturally-occurring.” [0043] Disclosed herein may be certain binding molecules, or antigen-binding fragments, variants, or derivatives thereof. Unless specifically referring to full-sized antibodies such as Atty. Dkt. No.64100-229640 naturally-occurring antibodies, the term "binding molecule" encompasses full-sized antibodies as well as antigen-binding fragments, variants, analogs, or derivatives of such antibodies, e.g., naturally-occurring antibody or immunoglobulin molecules or engineered antibody molecules or fragments that bind antigen in a manner similar to antibody molecules. [0044] As used herein, the term “binding molecule” refers in its broadest sense to a molecule that specifically binds an antigenic determinant. As described further herein, a binding molecule can comprise one of more “binding domains.” As used herein, a "binding domain" is a two- or three-dimensional polypeptide structure that cans specifically bind a given antigenic determinant, or epitope. A non-limiting example of a binding molecule is an antibody or fragment thereof that comprises a binding domain that specifically binds an antigenic determinant or epitope. Another example of a binding molecule is a bispecific antibody comprising a first binding domain binding to a first epitope, and a second binding domain binding to a second epitope. [0045] The terms "antibody" and "immunoglobulin" can be used interchangeably herein. An antibody (or a fragment, variant, or derivative thereof as disclosed herein comprises at least the variable domain of a heavy chain and at least the variable domains of a heavy chain and a light chain. Basic immunoglobulin structures in vertebrate systems are relatively well understood. See, e.g., Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed.1988). [0046] Binding molecules, e.g., antibodies or antigen-binding fragments, variants, or derivatives thereof include, but are not limited to, polyclonal, monoclonal, human, humanized, or chimeric antibodies, single chain antibodies, epitope-binding fragments, e.g., Fab, Fab' and F(ab')2, Fd, Fvs, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv), fragments comprising either a VL or VH domain, fragments produced by a Fab expression library. ScFv molecules are known in the art and are described, e.g., in US patent 5,892,019. Immunoglobulin or antibody molecules encompassed by this disclosure can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecule. [0047] By "specifically binds," it is meant that a binding molecule, e.g., an antibody or fragment, variant, or derivative thereof binds to an epitope via its antigen binding domain, and that the binding entails some complementarity between the antigen binding domain and the epitope. According to this definition, a binding molecule is said to "specifically bind" to an epitope when it binds to that epitope, via its antigen-binding domain more readily than it would bind to a random, Atty. Dkt. No.64100-229640 unrelated epitope. The term "specificity" is used herein to qualify the relative affinity by which a certain binding molecule binds to a certain epitope. For example, binding molecule "A" can be deemed to have a higher specificity for a given epitope than binding molecule "B," or binding molecule "A" can be said to bind to epitope "C" with a higher specificity than it has for related epitope "D." [0048] The term "polynucleotide" is intended to encompass a singular nucleic acid as well as plural nucleic acids, and refers to an isolated nucleic acid molecule or construct, e.g., messenger RNA (mRNA) or plasmid DNA (pDNA). A polynucleotide can comprise a conventional phosphodiester bond or a non-conventional bond (e.g., an amide bond, such as found in peptide nucleic acids (PNA)). The term "nucleic acid" refers to any one or more nucleic acid segments, e.g., DNA or RNA fragments, present in a polynucleotide. By "isolated" nucleic acid or polynucleotide is intended a nucleic acid molecule, DNA or RNA, which has been removed from its native environment. For example, a recombinant polynucleotide encoding a polypeptide subunit contained in a vector is considered isolated as disclosed herein. Further examples of an isolated polynucleotide include recombinant polynucleotides maintained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. Isolated RNA molecules include in vivo or in vitro RNA transcripts of polynucleotides. Isolated polynucleotides or nucleic acids further include such molecules produced synthetically. In addition, polynucleotide or a nucleic acid can be or can include a regulatory element such as a promoter, ribosome binding site, or a transcription terminator. [0049] Two DNA fragments (such as a polypeptide coding region and a promoter associated therewith) can be “operably associated” or “operably linked” if induction of promoter function results in the transcription of mRNA encoding the desired gene product and if the nature of the linkage between the two DNA fragments does not interfere with the ability of the expression regulatory sequences to direct the expression of the gene product or interfere with the ability of the DNA template to be transcribed. Thus, a promoter region would be operably associated with a nucleic acid encoding a polypeptide if the promoter was capable of effecting transcription of that nucleic acid. Two adjacent or overlapping coding sequences would be operable linked if both are capable of effective transcription. [0050] As used herein, a “non-naturally occurring” polynucleotide, or any grammatical variants thereof, is a conditional definition that explicitly excludes, but only excludes, those forms of the polynucleotide that are well-understood by persons of ordinary skill in the art as being Atty. Dkt. No.64100-229640 “naturally-occurring,” or that are, or that might be at any time, determined or interpreted by a judge or an administrative or judicial body to be, “naturally-occurring.” [0051] In certain embodiments, the polynucleotide or nucleic acid is DNA. In other embodiments, a polynucleotide can be RNA. [0052] A "vector" (also use interchangeably herein with “plasmid”) is nucleic acid molecule as introduced into a host cell, thereby producing a transformed host cell. A vector can include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector can encode and express a protein. A vector can also include one or more selectable marker gene and other genetic elements known in the art. [0053] A "transformed" cell, or a "host" cell, is a cell into which a nucleic acid molecule has been introduced by molecular biology techniques. As used herein, the term transformation encompasses those techniques by which a nucleic acid molecule can be introduced into such a cell, including transfection with viral vectors, transformation with plasmid vectors, and introduction of naked DNA by electroporation, lipofection, and particle gun acceleration. A transformed cell or a host cell can be a bacterial cell or a eukaryotic cell. [0054] The term “expression” as used herein refers to a process by which a gene produces a biochemical, for example, a polypeptide. The process includes any manifestation of the functional presence of the gene within the cell including, without limitation, gene knockdown as well as both transient expression and stable expression. It includes without limitation transcription of the gene into messenger RNA (mRNA), and the translation of such mRNA into polypeptide(s). If the final desired product is a biochemical, expression includes the creation of that biochemical and any precursors. Expression of a gene produces a "gene product." As used herein, a gene product can be either a nucleic acid, e.g., a messenger RNA produced by transcription of a gene, or a polypeptide that is translated from a transcript. Gene products described herein further include nucleic acids with post transcriptional modifications, e.g., polyadenylation, or polypeptides with post translational modifications, e.g., methylation, glycosylation, the addition of lipids, association with other protein subunits, proteolytic cleavage, and the like. [0055] As used herein the terms "treat," "treatment," or "treatment of" (e.g., in the phrase "treating a subject") refers to reducing the potential for disease pathology, reducing the occurrence of disease symptoms, e.g., to an extent that the subject has a longer survival rate or reduced discomfort. For example, treating can refer to the ability of a therapy when administered to a subject, to reduce disease symptoms, signs, or causes. Treating also refers to mitigating or Atty. Dkt. No.64100-229640 decreasing at least one clinical symptom and/or inhibition or delay in the progression of the condition and/or prevention or delay of the onset of a disease or illness. [0056] By "subject" or "individual" or "animal" or "patient" or “mammal,” is meant any subject, particularly a mammalian subject, for whom diagnosis, prognosis, or therapy is desired. Mammalian subjects include humans, domestic animals, farm animals, sports animals, and zoo animals, including, e.g., humans, non-human primates, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, bears, and so on. [0057] The term "pharmaceutical composition" or “therapeutic composition” refers to a preparation that is in such form as to permit the biological activity of the active ingredient to be effective, and that contains no additional components that are unacceptably toxic to a subject to which the composition would be administered. Such composition can be sterile. [0058] As used herein, a “sequon” refers to a specific sequence of amino acids consisting of amino acid residues for recognition and subsequent glycosylation by a specific oligosaccharyltransferase. [0059] As used herein, a “glycoconjugate” refers to a polypeptide that is covalently linked to a carbohydrate moiety. It is understood that the carbohydrate moiety can be a monosaccharide, oligosaccharide, or polysaccharide. For purposes of this disclosure, a “glycoconjugate” is a specific type of “bioconjugate” as referred to herein. In certain embodiments as would be recognized in the art, a glycoprotein is a glycoconjugate. Overview [0060] Conjugate vaccines, consisting of a polysaccharide linked to a protein, are lifesaving prophylactics. Traditionally, conjugate vaccines are manufactured using chemical methodologies. However, in vivo bacterial conjugations have emerged as manufacturing alternatives. In vivo conjugation (bioconjugation) is reliant upon an oligosaccharyltransferase (OTase) to attach polysaccharides to proteins. [0061] Even with the introduction and implementation of pneumococcal conjugate vaccines over the last two decades, ~1.5 million deaths are still attributed to S. pneumoniae each year. This is due in part to the 90+ serotypes of S. pneumoniae and the complex manufacturing methods required to synthesize pneumococcal conjugate vaccines. Together these factors hinder global distribution and development of broader, more protective variations of the vaccines. To expedite development and lower manufacturing costs, disclosed herein is a platform for developing and making conjugate vaccines, for example pneumococcal conjugate vaccines, using in vivo Atty. Dkt. No.64100-229640 conjugation. This streamlined process has the potential to complement existing manufacturing pipelines or completely bypass the dependency on chemical conjugation methodologies, enabling the production of a more comprehensive conjugate vaccines. [0062] Traditional, chemical conjugate vaccine synthesis is considered complex, costly, and laborious (Frasch, C.E. Vaccine 27, 6468-6470 (2009)) however, in vivo conjugation has been thoroughly progressing as a viable biosynthetic alternative (Huttner, A. et al. Lancet Infect Dis 17, 528-537 (2017)). These strides are best highlighted by the successes of GlycoVaxyn, (now LimmaTech Biologics AG an independent company with direct ties to GlaxoSmithKline), a clinical stage biopharmaceutical company with multiple bioconjugate vaccines in various phases of clinical trials, one of which (Flexyn2a) has completed a Phase 2b challenge study. Although GlycoVaxyn has been at the forefront of the in vivo conjugation revolution, the ability to efficiently glycosylate carrier/acceptor proteins with polysaccharides containing glucose (Glc) as the reducing end sugar has been elusive and, expectedly, has stymied the development of bioconjugate vaccines targeting multiple pathogens, including but not limited to, Streptococcus pneumoniae, Streptococcus agalactiae, Klebsiella pneumoniae, and Enterococcus. [0063] Protein glycosylation in bacteria is commonly mediated by oligosaccharyltransferases that transfer oligosaccharides en bloc from preassembled lipid-linked precursors to acceptor proteins. Natively, O-linking oligosaccharyltransferases usually transfer a single repeat unit of the O-antigen or capsular polysaccharide to the side chains of serine or threonine residues on acceptor proteins. Three major families of bacterial O-linking oligosaccharyltransferases have been described across diverse genera: PglL, PglS, and TfpO. TfpO enzymes are limited to transferring short oligosaccharides both in their native context and when heterologously expressed in glycoengineered E. coli. On the other hand, PglL and PglS enzymes can transfer long-chain polysaccharides when heterologously expressed. [0064] Given that bacteria from the genus Acinetobacter have been shown to carry genes encoding for TfpO, PglL, and PglS orthologs, the inventors were interested in searching for new O-linking oligosaccharyltransferases within the Moraxellaceae family to which Acinetobacter belongs. As the different types of O-linking oligosaccharyltransferases cannot be clearly differentiated based on primary amino acid sequence alone, definitive family assignment typically relies on functional assays. Nevertheless, TfpO proteins are typically smaller in size, around 420 – 460 amino acids, than PglS and PglL proteins that are approximately 525 – 600 amino acids. Moreover, the amino acid sequences of the pilins that O-linking oligosaccharyltransferases Atty. Dkt. No.64100-229640 glycosylate are also sufficiently different between oligosaccharyltransferase families and can be used to help identify to which family an oligosaccharyltransferase belongs. In the case of PglS only a single protein, the type IV pilin-like protein ComP, serves as a cognate acceptor in its natural context (Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041). The PglS sequon is flanked by two cysteine residues and contains a conserved internal serine residue that is the site of glycosylation (Harding, C. M., et al. (2019) Nature Communications 10, 891; Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203). In contrast, the minimum TfpO sequon is located at the C-terminus of the type IV pilin protein PilA (Horzempa, J., et al. (2006) Journal of Biological Chemistry 281, 1128-1136; Harvey, H., et al. (2009) Journal of Bacteriology 191, 6513-6524; Comer Jason, E., et al. (2002) Infection and Immunity 70, 2837-2845). [0065] Disclosed herein is the discovery and functional characterization of a novel family of bacterial O-linking oligosaccharyltransferases termed TfpM from Moraxellaceae bacteria. TfpM proteins are similar in size and sequence to TfpO enzymes but can transfer long-chain polysaccharides to acceptor proteins. Phylogenetic analyses demonstrate that TfpM proteins cluster in different clades than known bacterial oligosaccharyltransferases. Using a representative TfpM enzyme from Moraxella osloensis, it was determined that TfpM glycosylates a C-terminal threonine of its cognate pilin-like protein and the minimal sequon required for glycosylation was identified. It was demonstrated that TfpM has a broad substrate tolerance and can transfer diverse polysaccharides including those with reducing-end glucose, galactose, or 2-N-acetyl sugars. It was also shown that a TfpM-derived bioconjugate is immunogenic and elicits serotype-specific polysaccharide IgG responses in mice. Thus the glycan substrate promiscuity of TfpM and identification of the minimal TfpM sequon renders this enzyme a valuable additional tool for expanding the glycoengineering toolbox. Bioinformatic identification of a new class of OTase carried by bacteria from the Moraxellaceae family. [0066] To identify genes encoding O-linking oligosaccharyltransferases, the inventors first searched the NCBI genome and whole-genome shotgun contig sequence databases using the Basic Local Alignment Search Tool (BLAST) and the PglSADP1 amino acid sequence (SEQ ID NO: 1) as the query. SEQ ID NO: 1_ PglSADP1 amino acid sequence MNSIFKKIKNYTIVSGVFFLGSAFIIPNTSNLSSTLYKELIAVLGLLILLTVKSFDYKKILIPK NFYWFLFVIFIIFIQLIVGEIYFFQDFFFSISFLVILFLSFLLGFNERLNGDDLIVKKIAWIFIIV VQISFLIAINQKIEIVQNFFLFSSSYNGRSTANLGQPNQFSTLILITLFLLCYLREKNSLNNM Atty. Dkt. No.64100-229640 VFNILSFCLIFANVMTQSRSAWISVILISLLYLLKFQKKIELRRVIFFNIVFWTLVYCVPLLF NLIFFQKNSYSTFDRLTMGSSRFEIWPQLLKAVFHKPFIGYGWGQTGVAQLETINKSSTK GEWFTYSHNLFLDLMLWNGFFIGLIISILILCFLIELYSSIKNKSDLFLFFCVVAFFVHCLLE YPFAYTYFLIPVGFLCGYISTQNIKNSISYFNLSKRKLTLFLGCCWLGYVAFWVEVLDISK KNEIYARQFLFSNHVKFYNIENYILDGFSKQLDFQYLDYCELKDKYQLLDFKKVAYRYP NASIVYKYYSISAEMKMDQKSANQIIRAYSVIKNQKIIKPKLKFCSIEY [0067] To shorten the list of hits and reduce the likelihood of identifying very similar orthologs of PglSADP1, the search was further refined to candidates that had less than 50% amino acid sequence identity to PglSADP1. Some of the top hits from this refined list were proteins that were much more similar in size to TfpO proteins, but the upstream cognate pilin protein contained both a ComP disulfide-flanked sequon as well as a PilA-like sequon at the C-terminus. The first identified pilin- oligosaccharyltransferase pairs were encoded in two Acinetobacter species: A. parvus DSM 16617 and A. townerii ZZC-3 (Table 1). Table 1. Organisms and accession numbers for the TfpM enzymes and their associated pilins

Atty. Dkt. No.64100-229640

[0068] The two oligosaccharyltransferases from these strains were closely related, having >96% sequence identity. Intrigued by these findings, the inventors looked further and identified other strains in the Moraxellaceae family that carried genes encoding similar putative oligosaccharyltransferase/pilin pairs. While many genes like those in A. parvus DSM 16617 and A. townerii ZZC-3 were found, most of the associated pilin proteins encoded upstream of the oligosaccharyltransferase lacked a ComP-like sequon. The accession numbers and protein sizes for twenty of these putative oligosaccharyltransferases-pilin-like protein pairs are listed in Table 1. The inventors did not observe any homologues in species outside of the Moraxellaceae family and, to distinguish these different oligosaccharyltransferases from other known enzymes, have termed them TfpM proteins (‘M’ for Moraxellaceae). Given the similar size of TfpM proteins to known TfpO proteins, it was initially hypothesized that these genes encoded variants of TfpO-PilA pairs like those found in Acinetobacter and Pseudomonas (Harding, C. M., et al. (2015) Molecular Microbiology 96, 1023-1041). However, a multiple sequence alignment of the twenty TfpM proteins with known PglS, PglL, and TfpO proteins demonstrated that the former had less than 26% sequence identity to archetypal oligosaccharyltransferases. Analysis of the phylogenetic tree produced by the multiple alignment showed that TfpM proteins cluster in different clades than TfpO, PglS and PglL proteins (Figure 1A and Figure 2). In contrast, the pilin genes located Atty. Dkt. No.64100-229640 immediately upstream of tfpM did not cluster in discrete clades (Figure 3) and displayed overall higher identity to PilA proteins, namely between 37% and 60%. Based on the coding sequence, most of the associated pilin proteins belong to the type IV major pilin family with the exception of those from Acinetobacter sp. CIP102143 and Acinetobacter sp. CIP102637 which lack the characteristic type III signal sequence at the N-terminus (Giltner Carmen, L., et al. (2012) Microbiology and Molecular Biology Reviews 76, 740-772). TfpM orthologs glycosylate engineered pilin-fusion proteins with the pneumococcal serotype 8 capsular polysaccharide. [0069] Although similar in size to TfpO proteins, TfpM proteins are sufficiently different in terms of their amino acid sequence warranting further investigation. The inventors were particularly interested in determining if TfpM proteins could transfer only short oligosaccharides to acceptor proteins, like TfpO proteins. Of the twenty TfpM oligosaccharyltransferases listed in Table 1, the inventors selected 13 representatives from different clades to test for glycosylation activity in a glycoengineered E. coli strain (Harding, C. M., and Feldman, M. F. (2019) Glycobiology 29, 519-529; Feldman, M. F., et al. (2005) Proceedings of the National Academy of Sciences of the United States of America 102, 3016). Previously, the inventors developed a chimeric acceptor protein strategy consisting of the exotoxin A protein from Pseudomonas aeruginosa (EPA) fused to different-sized, soluble fragments of ComP (the natural substrate of PglS) (Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203). All type IV pilin-like proteins contain a conserved, N-terminal pilin signal sequence and membrane-anchoring domain that is not required for glycosylation but is essential for pilin stability. The fusion protein approach allows for the removal of the conserved N-terminal pilin signal sequence and membrane-anchoring domain and was used to determine the minimum sequon that PglS_ADP1 could recognize and still efficiently glycosylate (Harding, C. M., et al. (2019) Nature Communications 10, 891; Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203). The inventors adapted this approach and designed 13 synthetic double-stranded DNA blocks encoding an N-terminally truncated fragment of the upstream pilin gene and the downstream tfpM gene. In most of the strains carrying tfpM, the protein coding regions of the upstream pilin gene and the start codon of tfpM overlap by a single nucleotide. This genetic architecture was left intact in the expression constructs. The synthetic DNA blocks were designed such that, when cloned into an EPA expression vector using Gibson assembly, it placed the pilin coding region in frame with the C-terminus of EPA, creating a gene fusion that translated as a single protein with the tfpM gene immediately downstream (Figure 1B). The Atty. Dkt. No.64100-229640 truncated pilin fragments ranged from 113 to 140 amino acids in size. No purification tag was added to the C-terminus of the EPA-pilin fusion as previous studies with TfpO from P. aeruginosa 1244 reported that adding additional C-terminal residues after the serine residue prevented glycosylation (Horzempa, J., et al. (2006) Journal of Biological Chemistry 281, 1128-1136). Expression of the EPA-pilin and TfpM proteins was driven from an IPTG-inducible tac promoter on a pEXT20 plasmid (Dykxhoorn, D. M., et al. (1996) Gene 177, 133-136). The fusion protein was secreted into the periplasm using a DsbA signal sequence at the N-terminus of EPA. Oligos and primers used for the assemblies are listed in Table 2. Table 2. Primers and Oligos )

Atty. Dkt. No.64100-229640 )

[0070] Using this design, the inventors assessed the ability of the 13 TfpM proteins to transfer the Streptococcus pneumoniae capsular polysaccharide 8 (CPS8) glycan to their cognate pilin domain on the EPA-pilin fusions. The CPS8 repeat unit is a tetrasaccharide with a glucose at the Atty. Dkt. No.64100-229640 reducing end. Notably, PglS is thus far the only known oligosaccharyltransferase able to naturally transfer this glycan to acceptor proteins (Harding, C. M., et al. (2019) Nature Communications 10, 891). The 13 EPA-pilin fusions/TfpM expression vectors were transformed individually into E. coli SDB1 strains (Feldman, M. F., et al. (2005) Proceedings of the National Academy of Sciences of the United States of America 102, 3016) expressing the CPS8 glycan and assessed for protein glycosylation. The unglycosylated fusion proteins had expected masses ranging from 78.3 to 80.5 kDa. Several TfpM proteins were found to glycosylate their cognate EPA-pilin fusion with glycosylation appearing as higher molecular-weight laddering (gn) above the unglycosylated band (g0) (Figure 1C). Each higher-weight band represents the attachment of a glycan with one additional CPS8 repeat unit to the EPA-pilin protein. Glycosylation was readily observed in seven TfpM orthologs tested: Acinetobacter sp. YZSX-1-1, Acinetobacter sp. CIP102637, Acinetobacter sp. YH01026, A. junii 65, Acinetobacter sp. CIP102143, Acinetobacter sp. TUM15069, and M. osloensis 1202 (Figures 1C). Some of these extracts required higher exposure of the western blot to observe the glycosylation pattern (Figure 1D). As a control, the inventors generated a mutant of the M. osloensis TfpM protein gene with a single-residue change in a conserved histidine (His²⁸⁶) previously shown to be required for activity of wzy_C domain-containing enzymes (Figure 4) The wzy_C family pfam04932 is an “O-antigen ligase” domain present in membrane-bound enzymes that catalyze the transfer and covalent attached of lipid-linked oligosaccharide (liposaccharide) to lipid A or protein substrates. (Ruan, X., et al. (2012) Glycobiology 22, 288-299; Musumeci, M. A., et al. (2014) Glycobiology 24, 39-50). [0071] Glycosylation in SDB1 cell extracts expressing this H286A mutant alongside the EPA- pilin fusion protein and CPS8 glycan was not observed (Figure 1C) indicating that the glycosylation is due to the activity of the tfpM gene product and that His²⁸⁶ is required for catalytic activity and/or stability of this oligosaccharyltransferase. TfpMMo is an O-linking oligosaccharyltransferase that glycosylates the C-terminal threonine of its pilin substrate. [0072] Of the 13 TfpM-pilin pairs tested in the preceding experiment, those from M. osloensis 1202 and Acinetobacter sp. YH01026 exhibited the most efficient transfer of glycans of diverse sizes. Due to the slightly higher apparent stability of the pilin from M. osloensis FDAARGOS_1202 (1202 hereafter), the inventors chose the oligosaccharyltransferase from this organism as a representative for further characterization and have termed the enzyme TfpMMo (SEQ ID NO: 56). For clarity, reference is made to the intact, native M. osloensis 1202 pilin protein Atty. Dkt. No.64100-229640 as PilMo (SEQ ID NO: 57) and the N-terminally truncated fusion domain as PilMoH28 (SEQ ID NO: 58) throughout the text. Using TfpM_Mo, next was to identify the glycosylation site of Pil_MoH28 and thereby determine whether the enzyme acted like a TfpO protein, glycosylating the C-terminal amino acid of its cognate pilin acceptor, or if it was more like a PglL or PglS protein, glycosylating an internal residue. TfpO proteins transfer short oligosaccharides typically containing 3-6 sugars to the sidechain of the C-terminal serine residue of their cognate pilin. All except one of the cognate pilin proteins located immediately upstream a tfpM open reading frame ended in a C-terminal threonine residue, namely that from Psychrobacter sp. 72-O-c which ends in a serine. Based on this observation, as well as the size similarity of TfpM and TfpO proteins, the inventors hypothesized that TfpM enzymes also transfer glycans to a C-terminal residue and thus designed point mutants of the pilin to test this. Two mutants of the C-terminal pilin threonine (Thr¹⁶⁷) were generated, converting this residue to either serine or alanine and tested for glycosylation with CPS8. Whole cell E. coli extracts of the strains expressing the PilMoH28 mutants were probed using anti-EPA antibody in western blots. As seen in Figure 5, TfpMMo was also able to glycosylate EPA- Pil_MoH28 T167S, but no glycosylation was observed in the alanine mutant. These results indicate that the hydroxyl group on the sidechain of the Thr¹⁶⁷ or Ser¹⁶⁷ residue is likely the site of glycan attachment by TfpM_Mo. [0073] To confirm the site of pilin glycosylation by TfpM_Mo, CPS8-glycosylated EPA- PilMoH28 was partially purified and separated via SDS-PAGE analysis and Coomassie-stained the resolved glycoproteins. Gel slices corresponding to EPA- Pil_MoH28 glycosylated with 1 – 3 CPS8 repeat units were excised, digested with LysC, and analyzed for glycopeptides. Open searching- based analysis (Chick, J. M., et al. (2015) Nature Biotechnology 33, 743-749; Polasky, D. A., et al. (2020) Nature Methods 17, 1125-1132) of LysC-derived EPA-Pil_MoH28 peptides resulted in the identification of Hexose (Hex)-Hexuronic Acid (HexA)-modified ⁷⁶²FLPANCRGT⁷⁷⁰ peptide consistent with an incomplete monomer of the CPS8 glycan (HexHexAHex2). Higher-energy C- trap dissociation (HCD) analysis supports linkage of this disaccharide through the hexose residue and targeted Electron-Transfer/Higher-Energy Collision Dissociation (EThcD) analysis confirmed the attachment of HexHexA to the C-terminal threonine residue (Figure 6A and Figure 6B). While this MS analysis did not identify multimers of the CPS8 tetrasaccharide this is not surprising due to elongated glycoconjugates being extremely difficult to detect using peptide-centric LC-MS methods without the use of specialized chemical additives such as supercharging agents (Lin, C.- w., et al. (2016) Analytical Chemistry 88, 8484-8494). Nevertheless, the identification of a Atty. Dkt. No.64100-229640 disaccharide consistent with the partially completed CPS8 tetrasaccharide still supports the identity of the high molecular weight laddering as polymerized CPS8 tetrasaccharides. TfpMMo transfers polysaccharides containing glucose, galactose, or 2-N-acetyl monosaccharides at the reducing end. [0074] Next, it was sought to explore the range of polysaccharide substrates that TfpMMo can transfer in glycoengineered E. coli backgrounds. The inventors selected polysaccharides containing different reducing-end sugars, varying disaccharide sugar linkages near the reducing end, and/or were polymers composed of linear or branched repeat units. Four polysaccharides, E. coli O16 antigen, Salmonella enterica LT2 O-antigen, Klebsiella pneumoniae O2a antigen, and type III Group B Streptococcus capsular polysaccharide, in addition to the pneumococcal CPS8, were tested as glycan substrates for TfpMMo. The structures of all five repeat units tested are shown in Figure 7A (Liu, B., et al. (2020) FEMS Microbiology Reviews 44, 655-683; Curd, H., et al. (1998) Journal of Bacteriology 180, 1002-1007; Whitfield, C., et al. (1992) Journal of Bacteriology 174, 4913-4919; Pinto, V., and Berti, F. (2014) Journal of Pharmaceutical and Biomedical Analysis 98, 9-15; Geno, K. A., et al. (2015) Clinical Microbiology Reviews 28, 871-899). Each of the five polysaccharides were individually co-expressed with EPA-Pil_MoH28 and TfpM_Mo in E. coli SDB1 cells, induced, and grown for subsequent glycoprotein purification. Periplasmic extractions of SDB1 cells were partially purified using anion-exchange chromatography to remove any contaminating undecaprenol-pyrophosphate-linked polysaccharides that would convolute interpretation of the western blots. To demonstrate that the glycan-specific antibody signal observed in the western was indeed from glycosylated protein and not due to contaminating lipid- linked polysaccharide carryover from whole cell lysates, purified glycoproteins were split into two equal fractions, one half of which were digested with proteinase K prior to SDS-PAGE separation and western blotting. The western blots were probed using antisera specific to each polysaccharide and, separately, with anti-EPA antibody as all antibodies used in this experiment were from rabbits. As seen in Figure 7, TfpMMo was found to efficiently transfer all five different polysaccharides to the EPA-PilMoH28 protein. Digestion with proteinase-K eliminated both the anti-glycan (Figure 7B, Figure 7C, Figure 7D, Figure 7E, and Figure 7F) and anti-EPA (Figure 7G) signals in the western blots, confirming that the anti-glycan signals originated from protein-linked rather than contaminating lipid-linked polysaccharide. TfpMMo can glycosylate truncated PilMo 28 variants. Atty. Dkt. No.64100-229640 [0075] The prior experiments all utilized an N-terminally truncated variant of PilAMo that was 139 amino acids in length. To gain insights into the minimal features required for C-terminal pilin glycosylation by TfpMMo, a series of further-truncated variants of PilMoH28 and were generated and it was tested whether they could be glycosylated. The inventors first generated a 20 amino acid fragment of PilMo fused C-terminally to EPA via a flexible four-residue glycine linker, termed Pil20 (Figure 8A). This 20-amino acid fragment was chosen because it contains a disulfide loop (“DSL”) region that is conserved in many type IV pilins (Figure 9) (Horzempa, J., et al. (2006) Journal of Biological Chemistry 281, 1128-1136; Harvey, H., et al. (2009) Journal of Bacteriology 191, 6513- 6524). It is noted that this DSL corresponds to a different motif than the disulfide-flanked sequon present in ComP proteins (Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203). Based on sequence alignments with P. aeruginosa 1244 PilA, the DSL in M. osloensis pilin is formed by residues Cys¹⁴⁸ and Cys¹⁶⁴. As such, the inventors designed the sequence downstream of the glycine linker to start with Cys¹⁴⁸. The plasmid comprising the construct encoding EPA-Pil20 and TfpM was termed pVNM297. Glycosylation experiments with Pil20 revealed that it was able to be glycosylated by TfpMMo with CPS8 at similar levels to PilMoH28 (Figure 7B). To test whether the DSL region was required for glycosylation, several shorter variants were generated that lacked part of this feature. In these smaller constructs, the inventors also mutated Cys¹⁶⁴ to alanine to prevent the formation of non-natural disulfide linkages upon oxidation in the periplasm (Harvey, H., et al. (2009) Journal of Bacteriology 191, 6513-6524). 15-, 13-, and 10-amino acid PilMo variants were tested, with one variant of the 10-amino acid version containing an amino acid linker (e.g., GGGG linker) and one without. These constructs were termed Pil15, Pil13, Pil10L, and Pil10, respectively (Figure 8A). TfpMMo was able to glycosylate all four of these variants, albeit at lower levels than Pil₂₀ and Pil_MoH28 (Figure 5B). Both Pil₁₀ and Pil_10L were glycosylated comparably, indicating that the presence of the upstream glycine linker does not have a marked effect on TfpMMo activity. This linker was omitted from all subsequent constructs. Of the four proteins, the Pil13 was glycosylated notably worse than the others. [0076] From a sequence alignment, it was noted that each of the pilin proteins that were glycosylated by TfpM OTases have a conserved ‘P-A-N/E-C-R-G’ (SEQ ID NO: 200) motif found near the C-terminus, immediately upstream of the penultimate threonine residue (Figure 9). Given the presence of this feature in all glycosylated pilins, it was asked whether this motif was required for glycosylation by TfpM_Mo. The inventors fused a seven-amino acid variant, termed Pil₇, of Pil_Mo consisting of a similar motif (modified to ‘P-A-N-A-R-G-T’ (SEQ ID NO: 179), where the cysteine Atty. Dkt. No.64100-229640 is mutated to an alanine – bolded residue) to EPA and assessed glycosylation (Figure 8A). Thus, in certain embodiments, the glycosylated motif of the invention can comprise N, E, or A at the fourth position to the C-terminal end (e.g., corresponding to the N/E position of SEQ ID NO: 200 and second A position of SEQ ID NO: 179). The inventors also built stepwise single-amino acid truncations of this ‘P-A-N-A-R-G-T’ (SEQ ID NO: 179) sequence from seven to two amino acids and assessed the ability of TfpM to glycosylate these fragments with CPS8. The results showed that all variants except for Pil2 were glycosylated by TfpMMo at similar levels as Pil10 (Figure 8B). The overall glycosylation was again lower than for the Pil20 and PilMoH28. Glycosylated Pil2 was barely detectable though some trace laddering was visible at higher exposure of the western blot. This indicates that TfpMMo can glycosylate this variant but at markedly reduced levels than the construct with even one additional amino acid. From these results, it was concluded that TfpMMo can recognize and glycosylate a three-amino acid PilMo fragment when fused to the C-terminus of the heterologous EPA protein. Immunogenicity of a TfpMMo-derived GBSIII bioconjugate. [0077] Given that the EPA-Pil20 construct was efficiently glycosylated by TfpM, the inventors next assessed the ability of the EPA-Pil20 protein glycosylated with the type III capsular polysaccharide from Group B Streptococcus (GBSIII) for immunogenicity in a murine vaccination model. To aid in protein purification for these experiments, a plasmid expressing an N-terminally 6x-His tagged variant of the EPA-Pil20 carrier protein (pVNM291) was constructed. The His tag was added immediately downstream of the predicted N-terminal DsbA signal sequence cleavage site of EPA. pVNM291 was introduced into SDB1 cells expressing the GBSIII glycan and resulting bioconjugates purified using Nickel immobilized metal affinity chromatography (IMAC) followed by anion-exchange and size-exclusion chromatography on FPLC. Western blot and Coomassie staining of the SDS-PAGE resolved GBSIII-291 bioconjugate confirmed high molecular weight glycosylation of the EPA-Pil₂₀ protein with the GBSIII glycan (Figure 10A, Figure 10B, Figure 10C, and Figure 10D). Intact protein MS of the purified EPA-Pil20-GBSIII (“GBSIII-291”) conjugate supported a glycan:protein ratio of 20% (Figure 10E). Each dose was formulated to JVU[HPU * hN VM ;6E=== WVS`ZHJJOHYPKL' 5Z H JVU[YVS MVY [OLZL L_WLYPTLU[Z% \UNS`JVZ`SH[LK pVNM291-derived carrier protein (“291”) was purified from SDB1 cells without glycan plasmid and dosed at the same protein concentration as for the GBSIII bioconjugates. [0078] To test whether the TfpMMo-generated GBSIII-291 bioconjugate was immunogenic, 5- week-old female CD-1 mice were immunized. Mice received either placebo (unglycosylated 291 Atty. Dkt. No.64100-229640 carrier protein) or the GBSIII-291 bioconjugate at two-week intervals starting with a priming dose followed by two booster doses. All vaccines were formulated with a 1:9 ratio of Alhydrogel<sup>®</sup> 2% as an adjuvant. Serum was collected prior to each immunization and two weeks after the final booster. To determine the level of GBSIII-specific antibodies elicited, the inventors used an enzyme-linked immunosorbent assay (ELISA). All mice immunized with the bioconjugate GBSIII-291 were observed to express high levels of anti-GBSIII IgG antibodies, excluding a single mouse that exhibited a low anti-GBSIII IgG response, that was able to be boosted over the course of the immunization (Figure 10F). As expected, the GBSIII bioconjugate-vaccinated mice had increased GBSIII-specific IgG titers compared with the mock-vaccinated mice (291 alone, Figure 10F). Collectively, these data suggest that TfpMMo can produce bioconjugates capable of eliciting polysaccharide-specific IgG responses. TfpM<sub>Mo</sub> and PglS<sub>ADP1</sub> (PglL<sub>ComP</sub>) glycosylate a single protein engineered to contain sequons specific to each oligosaccharyltransferase. [0079] Lastly, the inventors wanted to determine if a protein engineered to contain sequons from two different OTase systems could be glycosylated by both OTases at each site. As such, an EPA fusion protein containing a sequon associated with TfpM as well as a sequon associated with PglS was constructed. To this end, the inventors engineered an EPA fusion protein containing a PglS sequon (CTGVTQIASGASAATTNVASAQC) (SEQ ID NO: 59) integrated between residues Ala<sup>489</sup> and Arg<sup>490</sup> as previously described (Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203) as well as the Pil<sub>20</sub> sequon (CGGTGTTVAAKFLPANCRGT) (SEQ ID NO: 60; same as Pil DSL) at the C-terminus (Figure 11A). As above, the construct was designed such that the open reading frame of the gene encoding for the EPA fusion and the start codon of tfpM overlapped by a single nucleotide. The open reading frame encoding for pglS from A. baylyi ADP1 was cloned 100 bp downstream of the tfpM open reading frame stop codon. This vector (pVNM337) was the introduced into E. coli SDB1 expressing the E. coli O16 antigen and tested for glycosylation via western blotting. To compare against proteins containing only a single sequon, the inventors individually introduced the following constructs into E. coli SDB1 expressing the O16 antigen: (i) EPA containing only the TfpM-associated Pil20 sequon (pVNM297) or (ii) EPA containing the PglS sequon integrated between residues Ala<sup>489</sup> and Arg<sup>490</sup> (pVNM167). To compare against a protein that has two sequons and can be di-glycosylated, the inventors also introduced an EPA construct containing the sequon integrated between residues Ala<sup>489</sup> and Arg<sup>490</sup> as well as between residues Glu<sup>548</sup> and Gly<sup>549</sup> (pVNM245) (EPA_PglS sequon 2X). Diagrams of these constructs are Atty. Dkt. No.64100-229640 shown in Figure 11A. As seen in Figure 11B, western blot analysis of EPA constructs containing only one sequon either from ComP or PilMo exhibited a glycoprofile around 100 kDa, indicative of a mono-glycosylation. The EPA construct containing two PglS sequons exhibited a predominantly mono-glycosylated profile around 100 kDa but also displayed a di-glycosylated population migrating around 150 kDa. Western blot analysis of the EPA fusion containing a sequon from TfpMMo and PglSADP1 displayed both mono- and di-glycosylated populations like that seen with construct pVNM245. It was concluded from these results that an acceptor protein can be glycosylated by two different OTase classes in one expression system. Glycoconjugates [0080] This disclosure provides for a glycoconjugate comprising an oligo- or polysaccharide covalently linked to an acceptor protein. In certain embodiments, the acceptor protein comprises or consists of a TfpM-associated pilin-like protein or glycosylation fragment thereof of this disclosure. In certain embodiments, the oligo- or polysaccharide is covalently linked to the pilin- like protein or glycosylation fragment thereof. And, in certain embodiments, the TfpM-associated pilin-like protein or glycosylation fragment thereof comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine. Further, in certain embodiments, the acceptor protein is a fusion protein comprising the TfpM- associated pilin-like protein or glycosylation fragment thereof translationally fused/linked to a heterologous amino acid sequence (e.g., a carrier protein) and the TfpM-associated pilin-like protein or glycosylation fragment thereof is the C-terminus-most sequence of the acceptor protein such that the acceptor protein comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine. Illustrative examples of carrier proteins include but are not limited to Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, or a fragment of any thereof. In certain embodiments, the TfpM-associated pilin-like protein or glycosylation fragment thereof is translationally fused/linked to a heterologous amino acid sequence/carrier protein via an amino acid linker. In certain embodiments, the oligo- or polysaccharide comprises a glucose at its reducing end. In certain embodiments, the glycoconjugate is immunogenic. [0081] In certain embodiments of a glycoconjugate of this disclosure, the acceptor protein comprises or consists of a full-length TfpM-associated pilin-like protein. In certain embodiments, the acceptor protein comprises or consists of a glycosylation fragment of a TfpM-associated pilin- like-protein that is less than a full-length TfpM-associated pilin-like protein. In certain Atty. Dkt. No.64100-229640 embodiments, the pilin-like-protein glycosylation fragment comprises a C-terminus serine or threonine residue. In certain embodiments, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end. In certain embodiments, the pilin- like-protein glycosylation fragment has a length of from 3 to 138 amino acids in length, has a length of from 10 to 138 amino acids in length, has a length of from 20 to 138 amino acids in length, 50 to 138 amino acids in length, has a length of from 100 to 138 amino acids in length, or has a length of from 116 to 138 amino acids in length. In certain embodiments, the pilin-like- protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 10 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, 50 to 139 amino acids in length, has a length of from 100 to 139 amino acids in length, or has a length of from 116 to 139 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 140 amino acids in length, has a length of from 10 to 140 amino acids in length, has a length of from 20 to 140 amino acids in length, 50 to 140 amino acids in length, has a length of from 100 to 140 amino acids in length, or has a length of from 116 to 140 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 3 to 21 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from any of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 to any of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In certain embodiments, the glycosylation fragment has a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. [0082] In certain embodiments of the glycoconjugate of this disclosure, the TfpM-associated pilin-like-protein or glycosylation fragment thereof is PilMo (SEQ ID NO: 57) or PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3 .1$ VY H WVS`WLW[PKL comprising at least comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, for example, wherein the C-terminus threonine is substituted with serine. In certain embodiments of the glycoconjugate of this disclosure, the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Atty. Dkt. No.64100-229640 PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100). In certain embodiments, the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment (e.g., C-terminus fragment) of any thereof and/or a variant wherein the C-terminus threonine is substituted with serine. And, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of the Pil_Mo pilin disulfide loop region (Pil_Mo_DSL, also referred to as Pil₂₀; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil₂₀ (SEQ ID NO: 60), Pil₁₉ (SEQ ID NO: 133), Pil₁₈ (SEQ ID NO: 134), Pil₁₇ (SEQ ID NO: 135), Pil₁₆ (SEQ ID NO: 136), Pil₁₅ (SEQ ID NO: 109), Pil₁₄ (SEQ ID NO: 137), Pil₁₃ (SEQ ID NO: 110), Pil₁₂ (SEQ ID NO: 138), Pil₁₁ (SEQ ID NO: 139), Pil₁₀ (SEQ ID NO: 112), Pil₉ (SEQ ID NO: 140), Pil₈ (SEQ ID NO: 141), Pil₇ (SEQ ID NO: 113), Pil₆ (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine. In certain of any of the above, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end Atty. Dkt. No.64100-229640 (RGT) or at least the last three amino acids from the pilin C-terminal end except that the C-terminus threonine is substituted with serine (RGS). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S[A] (SEQ ID NO: 183), Pil_19S[A] (SEQ ID NO: 184), Pil_18S[A] (SEQ ID NO: 185), Pil_17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil_12S[A] (SEQ ID NO: 191), Pil_11S[A] (SEQ ID NO: 192), Pil_10S[A] (SEQ ID NO: 193), Pil_9S[A] (SEQ ID NO: 194), Pil_8S[A] (SEQ ID NO: 195), Pil_7S[A] (SEQ ID NO: 196), Pil_6S[A] (SEQ ID NO: 197), Pil_5S[A] (SEQ ID NO: 198), or Pil_4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C- terminal end. [0083] In certain embodiments of the glycoconjugate of this disclosure, the acceptor protein can be glycosylated at two or more different positions. In certain embodiments, the acceptor protein can be glycosylated by at least two different OTase classes in one expression system. For example, in certain embodiments, the acceptor protein is a fusion protein and the fusion protein further comprises an additional glycosylation sequence (e.g., glycosylation fragment) of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus. For example, the other OTase can be PglB, PglL, or PglS. In certain embodiments, the additional glycosylation sequence is a sequence internal to the fusion protein (i.e., not the C-terminal or N-terminal most sequence). In certain embodiments, the additional glycosylation sequence is a sequence internal within the sequence of a carrier protein (e.g., Figure 11A). In certain embodiments, the additional glycosylation sequence is also covalently linked to an oligo- or polysaccharide. In certain embodiments, the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten Atty. Dkt. No.64100-229640 or more, fifteen or more, or twenty or more additional glycosylation sequences. In certain embodiments, the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five additional glycosylation sequences. In certain embodiments, the additional glycosylation sequences are identical. In certain embodiments, at least one additional glycosylation sequence differs from another. In certain embodiments, at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other. And, in certain embodiments, none of the additional glycosylation sequences are the same. In certain embodiments, the acceptor protein is a fusion protein and the fusion protein further comprises an internal glycosylation fragment of ComP in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C- terminus. In certain embodiments, the ComP glycosylation fragment is internal within the sequence of a carrier protein. In certain embodiments, the ComP glycosylation fragment is also covalently linked to an oligo- or polysaccharide. And, in certain embodiments, the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13. In certain embodiments, the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more ComP glycosylation fragments. In certain embodiments, the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five ComP glycosylation fragments. In certain embodiments, the ComP glycosylation fragments are identical. In certain embodiments, the ComP glycosylation fragments differ from each other. In certain embodiments, at least three, at least four, or at least five of the ComP glycosylation fragments all differ from each other. And, in certain embodiments, none of the ComP glycosylation fragments are the same. [0084] In certain embodiments of the glycoconjugate of this disclosure, the oligo- or polysaccharide covalently linked to the pilin-like protein or glycosylation fragment thereof has a size of at least three repeating units of oligo- or polysaccharide structure. In certain embodiments of the glycoconjugate of this disclosure, the oligo- or polysaccharide covalently linked to the pilin- like protein or glycosylation fragment thereof has a size of at least ten monosaccharides. [0085] In certain embodiments of the glycoconjugate of this disclosure, the oligo- or polysaccharide is produced by bacteria of the genus Streptococcus (for example, S. pneumoniae or Atty. Dkt. No.64100-229640 S. agalactiae) and the polysaccharide is capsular polysaccharide such as Ia, Ib, II, III, IV, V, VI, VII, VIII, or IX. [0086] In certain embodiments of the glycoconjugate of this disclosure, the oligo- or polysaccharide is produced by bacteria of the genus Klebsiella (for example, K. pneumoniae) and the polysaccharide is a capsular polysaccharide or O-antigen polysaccharide. [0087] In certain embodiments of the glycoconjugate of this disclosure, the oligo- or polysaccharide is produced by bacteria of the genus Salmonella and the polysaccharide is O- antigen polysaccharide. In certain embodiments, the bacteria is S. enterica and the S. enterica polysaccharide is a Group B O-antigen. [0088] In certain embodiments of the glycoconjugate of this disclosure, the glycoconjugate is produced in vivo, such as in a bacterial cell. In certain embodiments, the bacteria is Escherichia coli. In certain embodiments, the bacterium is from the genus Klebsiella. In certain embodiments, the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. In certain embodiments, the glycoconjugate is produced in a cell free system. [0089] In certain embodiments of the glycoconjugate of this disclosure, the bioconjugate is a conjugate vaccine that induces an immune response when administered to a subject. In certain embodiments, the immune response elicits long term memory (memory B and T cells), is an antibody response, and is optionally a serotype-specific antibody response. In certain embodiments, the antibody response is an IgG or IgM response. In certain embodiments, the antibody response is an IgG response, for example, an IgG1 response. And, in certain embodiments, the conjugate vaccine generates immunological memory in a subject administered the vaccine. Glycosylation fragments [0090] This disclosure provides for a pilin-like-protein glycosylation fragment comprising or consisting of an isolated fragment of a TfpM-associated pilin-like protein of this disclosure. In certain embodiments, the TfpM-associated pilin-like protein or pilin-like-protein glycosylation fragment comprises or consists of PilMo (SEQ ID NO: 57) or PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3 .1$ VY H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, for example, wherein the C-terminus threonine is substituted with serine. In certain embodiments of the glycoconjugate of this disclosure, the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 Atty. Dkt. No.64100-229640 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100). In certain embodiments, the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), Pil<sub>CIP102159</sub> (SEQ ID NO: 96), Pil<sub>junii-65</sub> (SEQ ID NO: 97), Pil<sub>YZS-X</sub> (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment (e.g., C-terminus fragment) of any thereof and/or a variant wherein the C-terminus threonine is substituted with serine. And, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of the Pil<sub>Mo</sub> pilin disulfide loop region (Pil<sub>Mo</sub>_DSL, also referred to as Pil<sub>20</sub>; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil<sub>20</sub> (SEQ ID NO: 60), Pil<sub>19</sub> (SEQ ID NO: 133), Pil<sub>18</sub> (SEQ ID NO: 134), Pil<sub>17</sub> (SEQ ID NO: 135), Pil<sub>16</sub> (SEQ ID NO: 136), Pil<sub>15</sub> (SEQ ID NO: 109), Pil<sub>14</sub> (SEQ ID NO: 137), Pil<sub>13</sub> (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil<sub>8</sub> (SEQ ID NO: 141), Pil<sub>7</sub> (SEQ ID NO: 113), Pil<sub>6</sub> (SEQ ID NO: 114), Pil<sub>5</sub> (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine. In certain Atty. Dkt. No.64100-229640 of any of the above, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end (RGT) or at least the last three amino acids from the pilin C-terminal end except that the C-terminus threonine is substituted with serine (RGS). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil<sub>17S[A]</sub> (SEQ ID NO: 186), Pil<sub>16S[A]</sub> (SEQ ID NO: 187), Pil<sub>15S[A]</sub> (SEQ ID NO: 188), Pil<sub>14S[A]</sub> (SEQ ID NO: 189), Pil<sub>13S[A]</sub> (SEQ ID NO: 190), Pil<sub>12S[A]</sub> (SEQ ID NO: 191), Pil<sub>11S[A]</sub> (SEQ ID NO: 192), Pil<sub>10S[A]</sub> (SEQ ID NO: 193), Pil<sub>9S[A]</sub> (SEQ ID NO: 194), Pil<sub>8S[A]</sub> (SEQ ID NO: 195), Pil<sub>7S[A]</sub> (SEQ ID NO: 196), Pil<sub>6S[A]</sub> (SEQ ID NO: 197), Pil<sub>5S[A]</sub> (SEQ ID NO: 198), or Pil<sub>4S[A]</sub> (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. [0091] In certain embodiments of the glycosylation fragment of this invention, the isolated fragment of a TfpM-associated pilin-like protein of this disclosure has a length of from 3 to 138 amino acids in length, has a length of from 10 to 138 amino acids in length, has a length of from 20 to 138 amino acids in length, 50 to 138 amino acids in length, has a length of from 100 to 138 amino acids in length, or has a length of from 116 to 138 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 10 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, 50 to 139 amino acids in length, has a length of from 100 to 139 amino acids in length, or has a length of from 116 to 139 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 140 amino acids in length, has a length of from 10 to 140 amino acids in length, has a length of from 20 to 140 amino acids in length, 50 to 140 amino acids in length, has a length of from 100 to 140 amino acids in length, or has a length of from 116 to 140 Atty. Dkt. No.64100-229640 amino acids in length. In certain embodiments, the glycosylation fragment has a length of has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from any of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 to any of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In certain embodiments, the glycosylation fragment has a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In certain embodiments, the pilin- like-protein glycosylation fragment comprises a C-terminus serine or threonine residue. In certain embodiments, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end. Fusion protein [0092] Provided for herein is a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof of this disclosure translationally fused/linked to a heterologous carrier protein such as, but not limited to, Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, or a fragment of any thereof. In certain embodiments, the TfpM-associated pilin-like protein or glycosylation fragment thereof is translationally fused/linked to a heterologous carrier protein via an amino acid linker. In certain embodiments, the pilin-like protein or glycosylation fragment comprises a C-terminus serine or threonine residue. In certain embodiments, the pilin-like protein or glycosylation fragment is the C-terminus-most sequence of the fusion protein. And, in certain embodiments, the fusion protein comprises a C-terminus serine or threonine residue. Further, in certain embodiments, the fusion protein is glycosylated by an oligo- or polysaccharide covalently linked to the C-terminus serine or threonine. Further, in certain embodiments, the fusion protein is glycosylated by an oligo- or polysaccharide comprising glucose at its reducing end covalently linked to the C-terminus serine or threonine. In certain embodiments, the glycosylated fusion protein is immunogenic. In certain embodiments, the glycosylated fusion protein is a conjugate vaccine. [0093] In certain embodiments of the fusion protein of this disclosure, the fusion protein comprises a full-length TfpM-associated pilin-like protein. In certain embodiments, the fusion protein comprises or consists of a glycosylation fragment of a TfpM-associated pilin-like-protein that is less than a full-length TfpM-associated pilin-like protein. In certain embodiments, the pilin- Atty. Dkt. No.64100-229640 like-protein glycosylation fragment comprises a C-terminus serine or threonine residue. In certain embodiments, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end. In certain embodiments, the pilin-like-protein glycosylation fragment has a length of from 3 to 138 amino acids in length, has a length of from 10 to 138 amino acids in length, has a length of from 20 to 138 amino acids in length, 50 to 138 amino acids in length, has a length of from 100 to 138 amino acids in length, or has a length of from 116 to 138 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 10 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, 50 to 139 amino acids in length, has a length of from 100 to 139 amino acids in length, or has a length of from 116 to 139 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 140 amino acids in length, has a length of from 10 to 140 amino acids in length, has a length of from 20 to 140 amino acids in length, 50 to 140 amino acids in length, has a length of from 100 to 140 amino acids in length, or has a length of from 116 to 140 amino acids in length. In certain embodiments, the glycosylation fragment has a length of has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from any of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 to any of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In certain embodiments, the glycosylation fragment has a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. [0094] In certain embodiments of the fusion protein of this disclosure, the pilin-like-protein glycosylation fragment comprises or consists of PilMo (SEQ ID NO: 57) or PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3.1$ VY H WVS`WLW[PKL JVTWYPZPUN H[ least comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, for example, wherein the C-terminus threonine is substituted with serine. In certain embodiments, the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 Atty. Dkt. No.64100-229640 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100). In certain embodiments, the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment (e.g., C-terminus fragment) of any thereof and/or a variant wherein the C-terminus threonine is substituted with serine. And, in certain embodiments, the pilin-like- protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148). Further, in certain embodiments, the pilin- like-protein glycosylation fragment comprises or consists of Pil₂₀ (SEQ ID NO: 60), Pil₁₉ (SEQ ID NO: 133), Pil₁₈ (SEQ ID NO: 134), Pil₁₇ (SEQ ID NO: 135), Pil₁₆ (SEQ ID NO: 136), Pil₁₅ (SEQ ID NO: 109), Pil₁₄ (SEQ ID NO: 137), Pil₁₃ (SEQ ID NO: 110), Pil₁₂ (SEQ ID NO: 138), Pil₁₁ (SEQ ID NO: 139), Pil₁₀ (SEQ ID NO: 112), Pil₉ (SEQ ID NO: 140), Pil₈ (SEQ ID NO: 141), Pil₇ (SEQ ID NO: 113), Pil₆ (SEQ ID NO: 114), Pil₅ (SEQ ID NO: 115), Pil₄ (SEQ ID NO: 116), or Pil₃ (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine. Further, in certain embodiments, the pilin- like-protein glycosylation fragment comprises or consists of Pil_20S (SEQ ID NO: 148), Pil_19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine. In certain of any of the above, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end (RGT) or at least the last three amino acids from the pilin C-terminal end except that the C-terminus threonine is substituted with serine (RGS). Further, in certain embodiments, Atty. Dkt. No.64100-229640 the pilin-like-protein glycosylation fragment comprises or consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil_13S[A] (SEQ ID NO: 190), Pil_12S[A] (SEQ ID NO: 191), Pil_11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil_6S[A] (SEQ ID NO: 197), Pil_5S[A] (SEQ ID NO: 198), or Pil_4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. [0095] In certain embodiments of the fusion protein of this disclosure, the fusion protein can be glycosylated at two or more different positions. In certain embodiments, the fusion protein can be glycosylated by at least two different OTase classes in one expression system. For example, in certain embodiments, the fusion protein further comprises a glycosylation sequence (e.g., glycosylation fragment) of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus. For example, the other OTase can be PglB, PglL, or PglS. In certain embodiments, the additional glycosylation sequence is a sequence internal to the fusion protein (i.e., not the C-terminal or N- terminal most sequence). In certain embodiments, the additional glycosylation sequence is a sequence internal within the sequence of a carrier protein (e.g., Figure 11A). In certain embodiments, the additional glycosylation sequence is also covalently linked to an oligo- or polysaccharide. In certain embodiments, the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more additional glycosylation sequences. In certain embodiments, the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more Atty. Dkt. No.64100-229640 than twenty, or more than twenty five additional glycosylation sequences. In certain embodiments, the additional glycosylation sequences are identical. In certain embodiments, at least one additional glycosylation sequence differs from another. In certain embodiments, at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other. And, in certain embodiments, none of the additional glycosylation sequences are the same. In certain embodiments, the fusion protein further comprises an internal glycosylation fragment of ComP in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus. In certain embodiments, the ComP glycosylation fragment is also covalently linked to an oligo- or polysaccharide. And, in certain embodiments, the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13. In certain embodiments, the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more ComP glycosylation fragments. In certain embodiments, the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five ComP glycosylation fragments. In certain embodiments, the ComP glycosylation fragments are identical. In certain embodiments, the ComP glycosylation fragments differ from each other. In certain embodiments, at least three, at least four, or at least five of the ComP glycosylation fragments all differ from each other. And, in certain embodiments, none of the ComP glycosylation fragments are the same. [0096] In certain embodiments of the fusion protein of this disclosure, the oligo- or polysaccharide covalently linked to the pilin-like protein or glycosylation fragment thereof has a size of at least three repeating units of oligo- or polysaccharide structure. In certain embodiments of the fusion protein of this disclosure, the oligo- or polysaccharide covalently linked to the pilin- like protein or glycosylation fragment thereof has a size of at least ten monosaccharides. [0097] In certain embodiments of the fusion protein of this disclosure, the oligo- or polysaccharide is produced by bacteria of the genus Streptococcus (for example, S. pneumoniae or S. agalactiae) and the polysaccharide is capsular polysaccharide such as Ia, Ib, II, III, IV, V, VI, VII, VIII, or IX. [0098] In certain embodiments of the fusion protein of this disclosure, the oligo- or polysaccharide is produced by bacteria of the genus Klebsiella (for example, K. pneumoniae) and the polysaccharide is a capsular polysaccharide or O-antigen polysaccharide. Atty. Dkt. No.64100-229640 [0099] In certain embodiments of the fusion protein of this disclosure, the oligo- or polysaccharide is produced by bacteria of the genus Salmonella and the polysaccharide is O- antigen polysaccharide. In certain embodiments, the bacteria is S. enterica and the S. enterica polysaccharide is a Group B O-antigen. [0100] In certain embodiments of the fusion protein of this disclosure, the glycosylated fusion protein is produced in vivo, such as in a bacterial cell. In certain embodiments, the bacteria is Escherichia coli. In certain embodiments, the bacterium is from the genus Klebsiella. In certain embodiments, the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. [0101] In certain embodiments of the fusion protein of this disclosure, the fusion protein is a vaccine that induces an immune response when administered to a subject. In certain embodiments, the immune response elicits long term memory (memory B and T cells), is an antibody response, and is optionally a serotype-specific antibody response. In certain embodiments, the antibody response is an IgG or IgM response. In certain embodiments, the antibody response is an IgG response, for example, an IgG1 response. And, in certain embodiments, the fusion protein generates immunological memory in a subject administered the fusion protein. Method of producing a glycoconjugate [0102] Provided for herein is a method of producing a glycoconjugate. In certain embodiments, the method occurs in vivo. In certain aspects, the glycoconjugate is produced in a cell free system. Examples of the use of a cell free system utilizing OTases other than TfpM can be found in WO2013/067523A1, which is in incorporated herein by reference. In certain embodiments, the method comprises covalently linking (conjugating) an oligo- or polysaccharide to an acceptor protein comprising or consisting of a TfpM-associated pilin-like protein or glycosylation fragment thereof using a TfpM oligosaccharyltransferase (OTase) of this disclosure. In certain embodiments, the pilin-like protein or glycosylation fragment comprises a C-terminus serine or threonine residue, the acceptor protein comprises a C-terminus serine or threonine residue, and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine residue of the acceptor protein. In certain embodiments, the oligo- or polysaccharide comprises a glucose at its reducing end. In certain embodiments, the acceptor protein is a fusion protein of this disclosure as described in detail elsewhere herein. And, in certain embodiments, the glycoconjugate is immunogenic. [0103] In certain embodiments of a method of producing a glycoconjugate of this disclosure or with any other compositions or methods disclosed herein, the TfpM OTase contains a wzy_C Atty. Dkt. No.64100-229640 superfamily domain that is defined by the .National Library of Science (NCBI) conserved protein domain family cl04850, an O-antigen ligase domain, and/or the TfpM OTase contains a wzy_C domain defined by the European Molecular Biology Laboratory (EMBL) European Bioinformatics Institute (EBI, EMBL-EBI) protein family (pfam) conserved protein domain family motif pfam04932, an O-antigen ligase domain, wherein pfam04932 is a protein domain family in the cl04850 superfamily protein domain. In certain embodiments, the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpM_YZS-X (SEQ ID NO: 76), TfpM_CIP102637 (SEQ ID NO: 77), TfpM_T-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81). In certain embodiments, the TfpM OTase is TfpM_Mo (SEQ ID NO: 56), TfpM_DSM16617 (SEQ ID NO: 63), TfpM_ZZC3 (SEQ ID NO: 64), TfpM_TUM15069 (SEQ ID NO: 65), TfpM_AI7 (SEQ ID NO: 66), TfpM_VE-C3 (SEQ ID NO: 67), TfpM_YH01026 (SEQ ID NO: 68), TfpM_CIP102143 (SEQ ID NO: 69), TfpM_AI40 (SEQ ID NO: 70), TfpM_F78 (SEQ ID NO: 71), TfpM_S71 (SEQ ID NO: 72), TfpM_ANC4282 (SEQ ID NO: 73), TfpM_CIP102159 (SEQ ID NO: 74), TfpM_junii-65 (SEQ ID NO: 75), TfpM_YZS-X (SEQ ID NO: 76), TfpM_CIP102637 (SEQ ID NO: 77), TfpM_T-3-2 (SEQ ID NO: 78), TfpM_BI730 (SEQ ID NO: 79), TfpM_A3K91 (SEQ ID NO: 80), or TfpM_72-O-c (SEQ ID NO: 81). In certain embodiments, the TfpM OTase is TfpM_Mo (SEQ ID NO: 56). [0104] In certain embodiments of a method of producing a glycoconjugate of this disclosure, the acceptor protein comprises or consists of a full-length TfpM-associated pilin-like protein. In certain embodiments, the acceptor protein comprises or consists of a glycosylation fragment of a TfpM-associated pilin-like-protein that is less than a full-length TfpM-associated pilin-like protein. In certain embodiments, the pilin-like-protein glycosylation fragment comprises a C-terminus serine or threonine residue. In certain embodiments, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end. In certain embodiments, the pilin-like-protein glycosylation fragment has a length of from 3 to 138 amino acids in length, has a length of from 10 to 138 amino acids in length, has a length of from 20 to 138 amino acids in length, 50 to 138 amino acids in length, has a length of from 100 to 138 amino acids in length, or has a length of from 116 to 138 amino acids in length. In certain embodiments, the pilin-like- Atty. Dkt. No.64100-229640 protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 10 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, 50 to 139 amino acids in length, has a length of from 100 to 139 amino acids in length, or has a length of from 116 to 139 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 140 amino acids in length, has a length of from 10 to 140 amino acids in length, has a length of from 20 to 140 amino acids in length, 50 to 140 amino acids in length, has a length of from 100 to 140 amino acids in length, or has a length of from 116 to 140 amino acids in length. In certain embodiments, the glycosylation fragment has a length of has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from any of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 to any of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In certain embodiments, the glycosylation fragment has a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. [0105] In certain embodiments of a method of producing a glycoconjugate of this disclosure, the pilin-like-protein glycosylation fragment comprises or consists of Pil_Mo (SEQ ID NO: 57) or Pil_Mo lacking amino acids corresponding to residues 1–28 (Pil_Mog+1% E9C =8 @A3 .1$ VY H polypeptide comprising at least comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, for example, wherein the C-terminus threonine is substituted with serine. In certain embodiments, the TfpM-associated pilin- like-protein is selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100). In certain embodiments, the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Atty. Dkt. No.64100-229640 PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment (e.g., C-terminus fragment) of any thereof and/or a variant wherein the C-terminus threonine is substituted with serine. And, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20 (SEQ ID NO: 60), Pil19 (SEQ ID NO: 133), Pil18 (SEQ ID NO: 134), Pil17 (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil₁₅ (SEQ ID NO: 109), Pil₁₄ (SEQ ID NO: 137), Pil₁₃ (SEQ ID NO: 110), Pil₁₂ (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil8 (SEQ ID NO: 141), Pil₇ (SEQ ID NO: 113), Pil₆ (SEQ ID NO: 114), Pil₅ (SEQ ID NO: 115), Pil₄ (SEQ ID NO: 116), or Pil₃ (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil_20S (SEQ ID NO: 148), Pil_19S (SEQ ID NO: 149), Pil_18S (SEQ ID NO: 150), Pil_17S (SEQ ID NO: 151), Pil_16S (SEQ ID NO: 152), Pil_15S (SEQ ID NO: 153), Pil_14S (SEQ ID NO: 154), Pil_13S (SEQ ID NO: 155), Pil_12S (SEQ ID NO: 156), Pil_11S (SEQ ID NO: 157), Pil_10S (SEQ ID NO: 158), Pil_9S (SEQ ID NO: 159), Pil_8S (SEQ ID NO: 160), Pil_7S (SEQ ID NO: 161), Pil_6S (SEQ ID NO: 162), Pil_5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine. In certain of any of the above, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end (RGT) or at least the last three amino acids from the pilin C-terminal end except that the C-terminus threonine is substituted with serine (RGS). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant Atty. Dkt. No.64100-229640 thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. [0106] In certain embodiments of a method of producing a glycoconjugate of this disclosure, the acceptor protein is a fusion protein and the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. In certain embodiments, the TfpM-associated pilin-like protein or glycosylation fragment thereof is translationally fused/linked to a heterologous carrier protein via an amino acid linker. [0107] In certain embodiments of a method of producing a glycoconjugate of this disclosure, the acceptor protein is a fusion protein and the method comprises glycosylating the acceptor protein at two or more different positions. In certain embodiments, the method comprises glycosylating the acceptor protein with at least two different OTase classes in one expression system. In certain embodiments, the fusion protein comprises two or more glycosylation sequences (e.g., glycosylation fragment) associated with at least two different OTases. Representative examples of OTases that may be used in combination include PglB, PglL, PglS, TfpO, and TfpM. One of ordinary skill in the art would recognize that OTases cannot be used together if they both require a glycosylation sequence (sequon) in the same position, e.g., both at the N-terminus or both at the C-terminus. For example, generally, TfpO and TfpM cannot be used together if they both need a sequon at the extreme C-terminus. For example, in certain non-limiting illustrative embodiments, the acceptor protein comprises a TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus in addition to an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase). In certain embodiments, the other OTase is PglB, PglL, and/or PglS. In certain embodiments, one or more glycosylation sequence is a sequence internal to the Atty. Dkt. No.64100-229640 fusion protein (i.e., not the C-terminal or N-terminal most sequence). In certain embodiments, the one or more glycosylation sequence is a sequence internal within the sequence of a carrier protein (e.g., Figure 11A). In certain embodiments, the additional glycosylation sequence is a sequence internal to the fusion protein (i.e., not the C-terminal or N-terminal most sequence). In certain embodiments, the additional glycosylation sequence is a sequence internal within the sequence of a carrier protein (e.g., Figure 11A). In certain embodiments, at least two different glycosylation sequences of two different OTase systems are covalently linked to an oligo- or polysaccharide. In certain embodiments, the TfpM-associated pilin-like-protein glycosylation fragment located at the fusion protein C-terminus and the additional glycosylation sequence are covalently linked to an oligo- or polysaccharide. In certain embodiments, the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more additional glycosylation sequences. In certain embodiments, the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five additional glycosylation sequences. In certain embodiments, the additional glycosylation sequences are identical. In certain embodiments, at least one additional glycosylation sequence differs from another. In certain embodiments, at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other. And, in certain embodiments, none of the additional glycosylation sequences are the same. For example, in certain embodiments, the method comprises further glycosylating an internal glycosylation fragment of ComP using a PglS OTase in addition to glycosylating the TfpM- associated pilin-like-protein glycosylation fragment located at its C-terminus. In certain embodiments, the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13. In certain embodiments, the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more ComP glycosylation fragments. In certain embodiments, the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five ComP glycosylation fragments. In certain embodiments, the ComP glycosylation fragments are identical. In certain embodiments, the ComP glycosylation fragments differ from each other. In certain embodiments, at least three, at least four, or at least five of the ComP glycosylation fragments all differ from each other. And, in certain embodiments, none of the ComP glycosylation fragments are the same. Atty. Dkt. No.64100-229640 [0108] In certain embodiments of a method of producing a glycoconjugate of this disclosure (e.g., conjugation) occurs in vivo in a host cell. In certain embodiments, the host cell is a bacterial cell. In certain embodiments, the conjugation occurs in in Escherichia coli. In certain embodiments, the conjugation occurs in a bacterium from the genus Klebsiella. In certain embodiments, the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. [0109] In certain embodiments of a method of producing a glycoconjugate of this disclosure, the method comprises culturing a host cell that comprises: (a) a genetic cluster encoding for the proteins required to synthesize the oligo- or polysaccharide; (b) a TfpM OTase; and (3) the acceptor protein. [0110] In certain embodiments of a method of producing a glycoconjugate of this disclosure, the method produces a conjugate vaccine. Additional embodiments [0111] Provided for herein is a host cell comprising (a) a genetic cluster encoding for the proteins required to synthesize an oligo- or polysaccharide; (b) a TfpM OTase of this disclosure; and (3) an acceptor protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof of this disclosure. In certain embodiments, the acceptor protein is a fusion protein. In certain embodiments, the host cell comprises a nucleic acid encoding the TfpM OTase. In certain embodiments, the host cell comprises a nucleic acid encoding the acceptor protein. And, in certain embodiments, the TfpM OTase and the acceptor protein are encoded by the same nucleic acid. [0112] Provided for herein is an isolated nucleic acid encoding the pilin-like-protein glycosylation fragment and/or the fusion protein of this disclosure. In certain embodiments, the nucleic acid is a vector. Also provided for is a host cell comprising such isolated nucleic acid of this disclosure. In certain embodiments, the host cell is a bacterial cell. In certain embodiments, the host cell is Escherichia coli. In certain embodiments, the host cell is from the genus Klebsiella. And, in certain embodiments, the host cell is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. [0113] Provided for herein is a composition comprising the conjugate vaccine or the fusion protein of this disclosure and an adjuvant and/or carrier. In certain embodiments, the composition is a pharmaceutical or therapeutic composition suitable for administration to a subject/patient. [0114] Provided for herein is a method of inducing a host immune response against a bacterial pathogen, the method comprising administering to a subject in need of the immune response an Atty. Dkt. No.64100-229640 effective amount of the conjugate vaccine, the fusion protein, or the composition comprising the conjugate vaccine or the fusion protein and an adjuvant and/or carrier of this disclosure. Treatment with pharmaceutical compositions comprising an immunogenic composition can occur separately or in conjunction with other treatments, as appropriate. An amount adequate to accomplish this is defined as “effective amount,” “effective dose,” or “unit dose.” Amounts effective for this use will depend on, e.g., the glycoconjugate composition, the manner of administration, the stage and severity of the disease being treated, the weight and general state of health of the patient, and the judgment of the prescribing physician. In some aspects, a priming dose is followed by a boosting dose over a period of time. In certain embodiments, the immune response is an antibody response. In certain embodiments, the immune response is selected from the group consisting of an innate response, an adaptive response, a humoral response, an antibody response, cell mediated response, a B cell response, a T cell response, cytokine upregulation or downregulation, immune system cross-talk, and a combination of two or more of said immune responses. In certain embodiments, the immune response is selected from the group consisting of an innate response, a humoral response, an antibody response, a T cell response, and a combination of two or more of said immune responses. [0115] Provided for herein is a method of preventing or treating a bacterial disease and/or infection in a subject comprising administering an effective amount to a subject in need thereof the conjugate vaccine, the fusion protein, or the composition comprising the conjugate vaccine or the fusion protein and an adjuvant and/or carrier of this disclosure. In certain embodiments, the subject is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a companion animal. In certain embodiments, the subject is livestock. In certain embodiments, the infection is a localized or systemic infection of skin, soft tissue, blood, or an organ, or is auto- immune in nature. In certain embodiments, the disease is pneumonia. In certain embodiments, the infection is a systemic infection and/or an infection of the blood. In certain embodiments, the conjugate vaccine, the fusion protein, or the composition is administered via intramuscular injection, intradermal injection, intraperitoneal injection, subcutaneous injection, intravenous injection, oral administration, mucosal administration, intranasal administration, or pulmonary administration. [0116] Provided for herein is a method of producing a pneumococcal conjugate vaccine against pneumococcal infection comprising: (a) isolating the glycoconjugate or a glycosylated fusion Atty. Dkt. No.64100-229640 protein of this disclosure; and (b) combining the isolated glycoconjugate or isolated glycosylated fusion protein with an adjuvant and/or carrier. [0117] Provided for herein is a glycoconjugate, glycosylated fusion protein, or conjugate vaccine, or a composition of any thereof of this disclosure for use in inducing a host immune response against a bacterial pathogen and/or preventing or treating a bacterial disease and/or infection in a subject. [0118] Provided for herein is a recombinant nucleic acid construct comprising a nucleotide sequence encoding a TfpM oligosaccharyltransferase (OTase) operably linked to at least one heterologous transcriptional regulatory sequence. In certain embodiments, the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpM_TUM15069 (SEQ ID NO: 65), TfpM_AI7 (SEQ ID NO: 66), TfpM_VE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpM_F78 (SEQ ID NO: 71), TfpM_S71 (SEQ ID NO: 72), TfpM_ANC4282 (SEQ ID NO: 73), TfpM_CIP102159 (SEQ ID NO: 74), TfpM_junii-65 (SEQ ID NO: 75), TfpM_YZS-X (SEQ ID NO: 76), TfpM_CIP102637 (SEQ ID NO: 77), TfpM_T-3-2 (SEQ ID NO: 78), TfpM_BI730 (SEQ ID NO: 79), TfpM_A3K91 (SEQ ID NO: 80), and/or TfpM_72-O-c (SEQ ID NO: 81). In certain embodiments, the TfpM OTase is TfpM_Mo (SEQ ID NO: 56), TfpM_DSM16617 (SEQ ID NO: 63), TfpM_ZZC3 (SEQ ID NO: 64), TfpM_TUM15069 (SEQ ID NO: 65), TfpM_AI7 (SEQ ID NO: 66), TfpM_VE-C3 (SEQ ID NO: 67), TfpM_YH01026 (SEQ ID NO: 68), TfpM_CIP102143 (SEQ ID NO: 69), TfpM_AI40 (SEQ ID NO: 70), TfpM_F78 (SEQ ID NO: 71), TfpM_S71 (SEQ ID NO: 72), TfpM_ANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpM_CIP102637 (SEQ ID NO: 77), TfpM_T-3-2 (SEQ ID NO: 78), TfpM_BI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81). In certain embodiments, the TfpM OTase is TfpMMo (SEQ ID NO: 56). In certain embodiments, the heterologous transcriptional regulatory sequence is a promotor sequence. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a TfpM-associated pilin- like protein or glycosylation fragment thereof of this disclosure or a fusion protein of this disclosure comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof operably linked to the nucleotide sequence encoding the a TfpM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a TfpM-associated pilin-like protein or glycosylation fragment thereof of this disclosure or a fusion protein of this disclosure comprising Atty. Dkt. No.64100-229640 a TfpM-associated pilin-like protein or glycosylation fragment thereof 5’ of and operably linked to the nucleotide sequence encoding the a TfpM OTase. In certain embodiments, the fusion protein of the construct also comprises a glycosylation sequence of an OTase other than TfpM, such as for PglB, PglL, PglS (e.g., ComP or a glycosylation fragment thereof). In certain embodiments, the coding sequence of the TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof is within 2, 5, 10, 20, 30, 40, or 50 nucleotides of the sequence encoding the TfpM OTase. In certain embodiments, the coding sequence of the TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof overlaps the operably linked nucleotide sequence encoding the a TfpM OTase. [0119] In certain embodiments, the TfpM-associated pilin-like protein comprises or consists of a full-length TfpM-associated pilin-like protein. In certain embodiments, the TfpM-associated pilin-like protein comprises or consists of a glycosylation fragment of a TfpM-associated pilin- like-protein that is less than a full-length TfpM-associated pilin-like protein. In certain embodiments, the pilin-like-protein glycosylation fragment comprises a C-terminus serine or threonine residue. In certain embodiments, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end. In certain embodiments, the pilin- like-protein glycosylation fragment has a length of from 3 to 138 amino acids in length, has a length of from 10 to 138 amino acids in length, has a length of from 20 to 138 amino acids in length, 50 to 138 amino acids in length, has a length of from 100 to 138 amino acids in length, or has a length of from 116 to 138 amino acids in length. In certain embodiments, the pilin-like- protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 10 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, 50 to 139 amino acids in length, has a length of from 100 to 139 amino acids in length, or has a length of from 116 to 139 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from 3 to 140 amino acids in length, has a length of from 10 to 140 amino acids in length, has a length of from 20 to 140 amino acids in length, 50 to 140 amino acids in length, has a length of from 100 to 140 amino acids in length, or has a length of from 116 to 140 amino acids in length. In certain embodiments, the glycosylation fragment has a length of has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length Atty. Dkt. No.64100-229640 of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length. In certain embodiments, the glycosylation fragment has a length of from any of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 to any of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In certain embodiments, the glycosylation fragment has a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. [0120] In certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of PilMo (SEQ ID NO: 57) or PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3 .1$ VY H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% /)"% 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, for example, wherein the C-terminus threonine is substituted with serine. In certain embodiments, the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), Pil_VE-C3 (SEQ ID NO: 86), Pil_YH01026 (SEQ ID NO: 87), Pil_CIP102143 (SEQ ID NO: 88), Pil_AI40 (SEQ ID NO: 89), Pil_F78 (SEQ ID NO: 90), Pil_S71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil_72-O-c (SEQ ID NO: 93), Pil_BI730 (SEQ ID NO: 94), Pil_A3K91 (SEQ ID NO: 95), Pil_CIP102159 (SEQ ID NO: 96), Pil_junii-65 (SEQ ID NO: 97), Pil_YZS-X (SEQ ID NO: 98), Pil_T-3-2 (SEQ ID NO: 99), and Pil_CIP102637 (SEQ ID NO: 100). In certain embodiments, the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), Pil_TUM15069 (SEQ ID NO: 84), Pil_AI7 (SEQ ID NO: 85), Pil_VE-C3 (SEQ ID NO: 86), Pil_YH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil_72-O-c (SEQ ID NO: 93), Pil_BI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment (e.g., C-terminus fragment) of any thereof and/or a variant wherein the C-terminus threonine is substituted with serine. And, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20 (SEQ ID NO: 60), Pil19 (SEQ ID NO: 133), Pil18 (SEQ ID Atty. Dkt. No.64100-229640 NO: 134), Pil17 (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil8 (SEQ ID NO: 141), Pil7 (SEQ ID NO: 113), Pil6 (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine. In certain of any of the above, the pilin-like-protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end (RGT) or at least the last three amino acids from the pilin C-terminal end except that the C-terminus threonine is substituted with serine (RGS). Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil_20[A] (SEQ ID NO: 166), Pil_19[A] (SEQ ID NO: 167), Pil_18[A] (SEQ ID NO: 168), Pil_17[A] (SEQ ID NO: 169), Pil_16[A] (SEQ ID NO: 170), Pil_15[A] (SEQ ID NO: 171), Pil_14[A] (SEQ ID NO: 172), Pil_13[A] (SEQ ID NO: 173), Pil_12[A] (SEQ ID NO: 174), Pil_11[A] (SEQ ID NO: 175), Pil_10[A] (SEQ ID NO: 176), Pil_9[A] (SEQ ID NO: 177), Pil_8[A] (SEQ ID NO: 178), Pil_7[A] (SEQ ID NO: 179), Pil_6[A] (SEQ ID NO: 180), Pil_5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. Further, in certain embodiments, the pilin-like-protein glycosylation fragment comprises or consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, Atty. Dkt. No.64100-229640 wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C- terminal end. [0121] In certain embodiments, the fusion protein is a fusion protein of this disclosure. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding an additional OTase as described elsewhere herein operably linked to the TpfM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding the additional OTase 3’ of and operably linked to the TpfM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding the additional OTase 5’ of and operably linked to the TpfM OTase. In certain embodiments, the coding sequence of the additional OTase is within 10, 20, 30, 40, 50, 75, or 100 nucleotides of the sequence encoding the TfpM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a PglS OTase 3’ operably linked to the TpfM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a PglS OTase 3’ of and operably linked to the TpfM OTase. In certain embodiments, the recombinant construct further comprises a nucleotide sequence encoding a PglS OTase 5’ of and operably linked to the TpfM OTase. In certain embodiments, the coding sequence of the PglS OTase is within 10, 20, 30, 40, 50, 75, or 100 nucleotides of the sequence encoding the TfpM OTase. [0122] Further provided for herein is a vector comprising the recombinant nucleic acid construct. And further provided for herein is a host cell comprising the recombinant nucleic acid construct or the vector. In certain embodiments, the host cell is a bacterial cell. In certain embodiments, the host cell is Escherichia coli. In certain embodiments, the host cell is from the genus Klebsiella. And, in certain embodiments, the host cell is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. Provided for herein is a method for producing a TfpM OTase comprising culturing the host cell, wherein said vector is an expression vector, and recovering the TfpM OTase. EXAMPLES Example 1. Cloning and Plasmid Assembly [0123] All primers and oligos used in this study are listed in Table 2. Working antibiotic concentrations used for liquid culture and in LB-agar plates were as follows: ampicillin (Amp), 100 µg/mL, kanamycin (Kan), 20 µg/mL, tetracycline (Tet), 10 µg/mL, spectinomycin (Sp), 50 µg/mL. For cloning the tfpM pilin-OTase genes, HiFi gblocks were ordered (Integrated DNA Technologies, IDT) designed with terminal 25 base pair overlaps for Gibson assembly with PCR- Atty. Dkt. No.64100-229640 linearized plasmid. The plasmid backbone for these fragments was amplified from a pEXT20 plasmid (Dykxhoorn, D. M., et al. (1996) Gene 177, 133-136) encoding the P. aeruginosa EPA gene under control of a tac promoter (pVNM57) (Knoot, C. J., et al. (2021) Glycobiology 31, 1192- 1203). The EPA gene has a deletion of residue E553 resulting in an inactivated toxin. The linearized plasmid was mixed separately with each of the

gBlocks and assembled using an NEBuilder HiFi DNA Assembly Kit (New England Biolabs, NEB). After assembly, the plasmids were transformed into E. coli Stellar cells (Takara Bio) by heat shock, out-grown for an hour at 37ºC, and plated on LB-agar supplemented with Amp. Individual colonies were picked and grown in LB media with appropriate antibiotic and plasmids isolated using a GeneJet Plasmid Miniprep Kit (Thermo Fisher). All plasmids were sequence-verified by Sanger sequencing (Genewiz). The plasmid expressing the M. osloensis *+)+ 9B5&BPSb+1 M\ZPVU HUK FMW?Mo was named pVNM227. To generate the Pil_Mo site-directed mutants, the inventors designed overlapping PCR primers that introduced the necessary codon changes in the pilin gene and amplified each fragment from pVNM227 plasmid. The resulting PCR product was DpnI-digested (NEB) for 30 min at 37ºC and gel-purified from agarose gel using a Pure-Link Gel Extraction Kit (Thermo Fisher). To insert the truncated pilin gene regions, complementary oligos with terminal 25 bp overlaps homologous to the pVNM227 PCR product were ordered. The oligos were re-suspended in purified water, mixed, and annealed together in thermocycler by heating to 98ºC for 5 min followed by a slow cooling to 4ºC at 0.1ºC/minute. The annealed oligos were diluted 1 to 5 in water and assembled with PCR-linearized pVNM227 using an NEBuilder HiFi DNA Assembly Kit (NEB). The resulting DNA was transformed into Stellar cells and plasmids isolated and verified as described above. The plasmid comprising the construct encoding EPA-Pil20 and TfpM was termed pVNM297. The N-terminally His-tagged EPA-Pil₂₀ variant was constructed by linearizing pVNM297 using PCR and using this fragment in Gibson assembly with complementary annealed oligos containing the 6xHis coding region and terminal homologous regions, resulting in pVNM291. pVNM167 was generated by digesting the previously described EPAiGTcc plasmid (Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203) with SalI. The purified SalI fragment was Gibson assembled with the pglS gene with its native 100 bp 5’ UTR amplified from A. baylyi ADP1 gDNA. pVNM245 was generated from pVNM167 template by separate PCR reactions to amplify products with overhangs for Gibson assembly: (i) the vector backbone with PglS and EPA with one iGT, (ii) the second iGT for integration between E548 and G549 and (iii) the C-terminus of EPA downstream of the iGT. The plasmid pVNM337 was created by amplifying tfpM from Atty. Dkt. No.64100-229640 pVNM291 with the primers EPA 3’ F1 and pglS-tfpM R1 and cloning the product into PCR- linearized pVNM167, which was amplified with pglS 5’ F1 and EPA 3’ R1. Phylogenetic trees for the TfpM and pilin proteins were generated using the phylogeny.fr server (on the world wide web at phylogeny.fr/) which uses MUSCLE, PhyML, and TreeDyn for sequence alignment, tree calculation, and image generation, respectively. Example 2. Expression of glycans and cloning of K. pneumoniae O2a glycan genes [0124] The S. pneumonia CPS8 glycan was expressed from plasmid pB8 (TetR) (Kay, E. J., et al. (2016) Open Biology 6, 150243), the Salmonella enterica LT2 glycan from plasmid pPR1347 (Kan^R) (Neal, B. L., et al. (1993) Journal of Bacteriology 175, 7115-7118), the E. coli O16 wbbL gene from plasmid pMF19 (Sp^R) (Feldman, M. F., et al. (2005) Proceedings of the National Academy of Sciences of the United States of America 102, 3016) and the GBSIII glycan was from a pBBR1MCS2 derivative (Duke, J. A., et al. (2021) ACS Infectious Diseases 7, 3111-3123). Bioconjugation with the K. pneumoniae O2a O-antigen has not previously been reported. To clone the genes that encode the machinery required to synthesize the O2a glycan, PCR was used to amplified the wzm, wzt, wbbM, glf, wbbN, and wbbO genes (Clarke, B. R., et al. (2018) Journal of Biological Chemistry 293, 4666-4679)(44) from K. pneumoniae strain NTUH K2044 genomic DNA. K. pneumoniae was cultured to saturation in LB media overnight and genomic DNA was isolated using a Wizard Genomic DNA Purification Kit (Promega). The plasmid backbone for the O2a cluster was amplified from plasmid pBBR1MCS2 (Kan^R) (Kovach, M. E., et al. (1995) Gene 166, 175-176). Primers for these reactions are listed in Table 2 The PCR products from these reactions were assembled using Gibson assembly with a NEBuilder HiFi DNA Assembly Kit (NEB). Stellar cells were transformed and plasmid isolated and verified as described in the previous section. Example 3. Bioconjugation and western blots [0125] E. coli strains used the bioconjugation experiments were either SDB1 or CLM24 (Feldman, M. F., et al. (2005) Proceedings of the National Academy of Sciences of the United States of America 102, 3016). SDB1 is a W3110 E. coli derivative with mutations in the genes encoding for WecA, the glycosyltransferase that initiates synthesis of the endogenous E. coli O16 antigen and WaaL, the enzyme transferring Und-PP linked glycans to Lipid A-core saccharide to produce LPS. CLM24 is a W3110 derivative with only a deletion of waaL. Elimination of these genes prevents crosstalk of the heterologous bioconjugation system and endogenous E. coli glycosylation pathways. To prepare E. coli strains for bioconjugation, the inventors electroporated Atty. Dkt. No.64100-229640 plasmids using competent cells prepared as previously described (Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203) followed by out-growth at 37ºC in SOB media. The cells were plated on LB-agar with appropriate antibiotics. The next day, 8 – 10 colonies were picked and inoculated into LB or TB with antibiotics and grown overnight while shaking at 30ºC. The next morning, starter cultures were inoculated into either 30 mL media in a 125 mL Erlenmeyer flask or 1 L media in a 2 L flask to a starting optical density at 600 nm (OD600) of 0.05. Cultures were grown while shaking at 175 RPM until the OD600 reached 0.4 – 0.6 at which point the cultures were induced with 1 mM IPTG. All bioconjugation experiments were performed at 30 ºC unless otherwise noted. After overnight induction, amounting to 20 – 24 hours total growth, the OD600 was measured, and 0.5 OD units of cells pelleted for analysis. [0126] The cell pellets were suspended in 100 µl 1X Laemmli Buffer (Biorad) and boiled for 10 min at 100ºC. The boiled samples were briefly centrifuged at 10,000 rcf and equivalent amounts, normalized to the same OD600 per lane, loaded for SDS-PAGE separation on a 7.5% Mini-Protean TGX gel (Biorad). Proteins were transferred to a nitrocellulose membrane using a semi-dry electrode system and blocked with Intercept Blocking Buffer (Li-Cor) for one hour. The membrane was incubated with primary antibodies in 1:1 blocking and TBST for 45 min. For protein detection, commercial rabbit anti-EPA and mouse anti-6xHis antibodies (Millipore-Sigma) were used. Rabbit glycan antibodies for CPS8, GBSIII, and O16 were purchased from SSI Diagnostica. K. pneumoniae rabbit O2a antibodies were a generous gift from Prof. Chris Whitfield (University of Guelph) (Clarke, B. R., et al. (2018) Journal of Biological Chemistry 293, 4666-4679). Salmonella Group B rabbit antibodies were purchased from BD. Primary incubation was followed by three washes with TBST buffer totaling 15 min The membranes were then incubated with secondary antibodies IRDye 680RD goat anti-mouse and/or IRDye 800CW goat anti-rabbit (Li-Cor) in 1:1 blocking buffer and TBST for 30 min. After a final 15-min TBST wash, the membranes were imaged using a Li-Cor Odyssey CLx. Example 4. Lys-C digestion of recombinant M. osloensis PilMob+1 [0127] In-gel digestion was accomplished according to the protocol of Shevchenko et al (Shevchenko, A., et al. (2006) Nat Protoc 1, 2856-2860) with minor modifications. Gel-separated NS`JVZ`SH[LK 9B5&BPSb+1 ^HZ L_JPZLK HUK KLZ[HPULK ^P[O KLZ[HPUPUN ZVS\[PVU #.) T? @<4HCO3, 50% ethanol) twice for 10 min at room temperature with shaking at 750 RPM. The destained band was then dehydrated with 100% ethanol for 10 min and dried by vacuum-centrifugation for 10 min before being rehydrated in 10 mM DTT in 50 mM NH4HCO3. Reduction was carried out for 60 Atty. Dkt. No.64100-229640 min at 56°C after which the gel band was dehydrated twice with 100% ethanol for 10 min to remove the remaining reduction buffer. The reduced sample was then sequentially alkylated with 55 mM iodoacetamide in 50 mM NH4HCO3 for 45 min at RT in the dark. The alkylated sample was then washed four times for 10 min with 50 mM NH4HCO3 followed by 100% ethanol, followed by 50 mM NH4HCO3 followed by 100% ethanol before being dried by vacuum-centrifugation. The dried alkylated sample was then rehydrated with 20 ng/µl Lys-C endoprotease (Wako Chemicals) in 40 mM NH4HCO3 at 4°C for 1hr. Excess Lys-C was removed, gel pieces were covered in 40mM NH4HCO3 and incubated overnight at 37°C. Peptides were concentrated and desalted using C18 stage tips (Ishihama, Y., et al. (2006) J Proteome Res 5, 988-994; Rappsilber, J., et al. (2007) Nat Protoc 2, 1896-1906) then eluted in buffer B (0.5% acetic acid, 80% acetonitrile (ACN)) before being dried and stored at -20°C prior to LC-MS analysis. Example 5. Analysis of recombinant M. osloensis Pil_Mob+1 \ZPUN YL]LYZLK WOHZL >7&?E [0128] The C18-concentrated digest was resuspended in buffer A* (0.1% TFA, 2% ACN) and separated using a two-column chromatography set up comprising a PepMap100 C₁₈20 mm x 75 hT [YHW HUK H BLW?HW 7₁₈ .)) TT _ 0. hT HUHS`[PJHS JVS\TU #FOLYTV :PZOLY EJPLU[PMPJ$' FOL ZHTWSL ^HZ JVUJLU[YH[LK VU[V [OL [YHW JVS\TU H[ . hS(TPU\[L \ZPUN )'*" MVYTPJ HJPK #:5$ MVY . min and infused into an Orbitrap Fusion™ Lumos™ Tribrid™ Mass Spectrometer equipped with a FAIMS Pro interface (Thermo Fisher Scientific) at 300 nl/min via the analytical column using a Dionex Ultimate 3000 UPLC (Thermo Fisher Scientific) by altering the concentration of buffer A (2% DMSO, 0.1% FA) and buffer B (78% ACN, 2% DMSO and 0.1% FA). Identification of potential glycopeptides utilized a 140-minute analytical run while targeted analysis utilized a 60- minute run. Within the identification analytical run the buffer composition was altered from 3% buffer B to 28% buffer B over 120 min, 28% buffer B to 40% buffer B over 9 min, 40% buffer B to 100% buffer B over 3 min, then the composition was held at 100% buffer B for 2 min, and then dropped to 3% buffer B over 2 min and held at 3% buffer B for another 8 min The Lumos™ Mass Spectrometer was operated in a stepped FAIMS data-dependent mode at three different FAIMS CVs, -25, -45 and -65 as previously described (Ahmad Izaham, A. R., et al. (2021) J Proteome Res 20, 599-612), switching between the acquisition of a single Orbitrap MS scan (60k resolution) every 1.5 sec followed by Orbitrap HCD scans (maximum fill time 120 milliseconds, AGC 2 x 10<sup>5</sup> with a resolution of 30k for Orbitrap MS-MS scans and an NCE of 25, 30,45) at each of the three FAIMS CVs. For targeted characterization of PilMo glycopeptides an analytical run altering the buffer composition from 3% buffer B to 15% buffer B over 30 min, 15% buffer B to 30% buffer Atty. Dkt. No.64100-229640 B over 10 min, 30% buffer B to 80% buffer B over 5 min, then the composition was held at 100% buffer B for 5 min, and then dropped to 3% buffer B over 1 min and held at 3% buffer B for another 9 min was undertaken. Parallel reaction monitoring using HCD (maximum fill time 250 ms, AGC 2.5 x 10<sup>5</sup> with a resolution of 30k for the Orbitrap MS-MS scan and an NCE of 15, 30,35) and EThcD (maximum fill time 250 ms, AGC 2.5 x 10<sup>5</sup> with a resolution of 30k for the Orbitrap MS- MS scan and ETD reaction times controlled using calibration charge dependent ETD parameters (Rose, C. M., et al. (2015) J Am Soc Mass Spectrom 26, 1848-1857) of the +2 charge state of the HexHexA modified glycopeptide <sup>762</sup>FLPANCRGT<sup>770</sup> (687.2972 m/z) was undertaken with a FAIMS CV of -45. Example 6. Open searching of PilMob+1 HUK [OL HUUV[H[PVU VM <L_<L_5&TVKPMPLK 7&[LYTPUHS peptide [0129] The identification of Pil<sub>Mo</sub> glycosylation events was accomplished using open database searching as previously described (Lewis, J. M., et al. (2021) J Vis Exp,). Briefly, datafiles were processed with MSfragger 3.4 (Polasky, D. A., et al. (2020) Nat Methods 17, 1125-1132; Kong, A. T., et al. (2017) Nat Methods 14, 513-520) in FragPipe (version 17.1) searching against the Moraxella osloensis Pil<sub>Mo</sub> sequence (NCBI Accession: WP_156627541.1). Searches were undertaken using “Lys-C” enzyme specificity with carbamidomethylation of cysteine as a fixed modification and oxidation of methionine as a variable modification with a maximum of 2 missed cleavages allowed. To enable identification of potential glycosylation events a mass tolerance, referred to as a delta mass, of 0 to 2000 Da was allowed. Delta masses observed on the C-terminal peptide <sup>762</sup>FLPANCRGT<sup>770</sup> (SEQ ID NO: 61) were manually inspected to identify potential glycosylation events. Parallel reaction monitoring results corresponding to the HexHexA-modified glycopeptide <sup>762</sup>FLPANCRGT<sup>770</sup> (SEQ ID NO: 62) was manually extracted using the Freestyle Viewer (1.7 SP1, Thermo Fisher Scientific) the MS/MS data annotated using the Interactive Peptide Spectral Annotator (Brademan, D. R., et al. (2019) Mol Cell Proteomics 18, S193-S201) (on the world wide web at interactivepeptidespectralannotator.com/PeptideAnnotator.html). Annotation of spectra allowed for the modification of the terminal T residue with HexHexA (338.0849 Da) as well as Hex (162.0528 Da). The resulting MS data and search results have been deposited into the PRIDE ProteomeXchange Consortium repository (Perez-Riverol, Y., et al. (2019) Nucleic Acids Res 47, D442-D450; Perez-Riverol, Y., et al. (2015) Proteomics 15, 930- 949) and can be accessed with the identifier PXD033468. Atty. Dkt. No.64100-229640 Example 7. Bioconjugate protein purification [0130] Cells for protein purification were grown in 1 L TB media and bioconjugates isolated using an osmotic shock protocol. After overnight growth and induction, the cells were pelleted by centrifugation, and washed in 0.9% NaCl. The washed cell pellets were suspended in 200 mM Tris- HCl pH 8.5, 100 mM EDTA, 25% sucrose and incubated while rolling for 30 min at 4ºC. Cells were pelleted by centrifugation at 4,700 rcf for 30 min and the resulting pellet suspended in 20 mM Tris-HCl pH 8.5 and incubated while rolling for 45 min at 4ºC. The suspension was centrifuged for 30 min at 18,000 rcf. The supernatant containing the periplasmic fraction was concentrated and LP[OLY SVHKLK KPYLJ[S` VU HU :B>7 HUPVU&L_JOHUNL JVS\TU VY% MVY <PZ&[HNNLK 9B5&BPSb+1 bioconjugates, purified using Nickel IMAC as previously described (Knoot, C. J., et al. (2021) Glycobiology 31, 1192-1203). The periplasmic extract or IMAC eluate was concentrated and buffer-exchanged into 20 mM Tris-HCl pH 8.0, filtered through a 0.2 µm PES filter then loaded on an Äkta pure FPLC instrument (Cytiva) equipped with a SOURCE 15Q 4.6/100 PE anion- exchange column (Cytiva). The bioconjugates were eluted at 2 mL/min using a stepwise gradient with buffer A (20 mM Tris pH 8) and buffer B (20 mM Tris pH 8, 1 M NaCl) from 0% B to 25% in 5% increments at 10 column volumes for each concentration. Bioconjugates for immunization were further purified using a Superdex 200 Increase 10/300 GL column. The concentrated bioconjugates pooled from the anion-exchange column were loaded on a pre-equilibrated Superdex 200 column in PBS buffer and eluted at a flow rate of 0.75 mL/min Fractions containing the purified bioconjugates were pooled, concentrated, and frozen at -80ºC for storage. Protein concentrations for immunization and western blots were determined using a Pierce BCA Protein Assay kit (Thermo Fisher). The ratio of polysaccharide to protein calculate for vaccine dosing was determined using the method described in Duke et al. (Duke, J. A., et al. (2021) ACS Infectious Diseases 7, 3111-3123). Example 8. Murine immunization [0131] All murine immunizations followed ethical regulations for animal testing and research. Experiments were carried out at Washington University School of Medicine in St. Louis according to the institutional guidelines and received approval from the Institutional Animal Care and Use Committee at Washington University in St. Louis. Five-week-old female CD-1 outbred mice #7OHYSLZ DP]LY >HIVYH[VYPLZ$ ^LYL Z\IJ\[HULV\ZS` PUQLJ[LK ^P[O *)) h> VM H ]HJJPUL MVYT\SH[PVU VU KH`Z )% *-% HUK +1' FOL ]HJJPUH[PVU NYV\WZ ^LYL +2* HSVUL #. hN WYV[LPU$ HUK ;6E===&+2* #. hN WYV[LPU% * hN WVS`ZHJJOHYPKL$' ?PJL OHK ZLYH JVSSLJ[LK VU KH`Z )% *-% +1% HUK -+' 5SS ]HJJPULZ Atty. Dkt. No.64100-229640 were formulated with Alhydrogel^® +" HS\TPU\T O`KYV_PKL NLS #=U]P]V;LU$ H[ H *32 YH[PV #.) h> ]HJJPUL [V .'. h> HS\T PU --'. h> *_ Z[LYPSL WOVZWOH[L I\MMLYLK ZHSPUL$' Example 9. Enzyme-linked immunosorbent assay (ELISA) [0132] IgG kinetic titers were determined using enzyme-linked immunosorbent assay (ELISA). Briefly, 96-well plates (TRP Immunomaxi plates) were coated in triplicate overnight with approximately 10<sup>6</sup> CFU/100 qL of glycoengineered E. coli expressing the GBSIII capsular polysaccharide in sodium carbonate buffer. The coating E. coli strain was grown the same as referenced above and after overnight induction to induce GBSIII expression was washed and diluted to coat plates. Wells were blocked with 1% BSA in PBS and washed with 0.05% PBS- Tween (PBST), all subsequent washes were the same. Serum from mice was diluted to 1:100 and added to wells for 1 hr at room temperature then washed. Total IgG titers were detected by HRP conjugated anti-mouse IgG (GE Lifesciences, 1:5000 dilution) added to wells for 1 hour at room [LTWLYH[\YL' 5M[LY ^HZOPUN% WSH[LZ ^LYL KL]LSVWLK \ZPUN ,%,f%.%.f [L[YHTL[O`S ILUaPKPUL #F?6$ substrate (Biolegend) and stopped with 2 N H2SO4. The optical densities were determined at 450 nm using a microplate reader (Bio-Tek). Total IgG product was determined using IgG standards to generate a standard curve for data fitting. Standard wells were coated with IgG in sodium carbonate buffer and treated the same as sample wells thereafter. All wells were normalized to blank wells that were treated the same as all samples wells minus receiving the primary mouse sera. Significance was determined using Mann-Whitney nonparametric test with P < 0.05. ***** [0133] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents. [0134] Certain embodiments of the present disclosure can be defined in any of the following numbered paragraphs: 1. A glycoconjugate comprising an oligo- or polysaccharide covalently linked to an acceptor protein, wherein the acceptor protein comprises or consists of a TfpM-associated pilin-like protein or glycosylation fragment thereof and the oligo- or polysaccharide is covalently linked to the TfpM-associated pilin-like protein or glycosylation fragment thereof, and wherein the TfpM-associated pilin-like protein or glycosylation fragment thereof comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine; Atty. Dkt. No.64100-229640 optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin-like protein C-terminal end; optionally, wherein the acceptor protein is a fusion protein comprising the TfpM- associated pilin-like protein or glycosylation fragment thereof translationally fused to a heterologous carrier protein and the TfpM-associated pilin-like protein or glycosylation fragment thereof is the C-terminus-most sequence of the acceptor protein such that the acceptor protein comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine; optionally, wherein the oligo- or polysaccharide comprises a glucose at its reducing end; and/or optionally, wherein the glycoconjugate is immunogenic. 2. The glycoconjugate of Paragraph 1, wherein the TfpM-associated pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 3. The glycoconjugate of Paragraph 1 or 2, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) Pil_Mo (SEQ ID NO: 57), (ii) Pil_Mo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3.1$% VY #PPP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ .)"% /)"% 0)"% 0."% 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein comprises a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: Atty. Dkt. No.64100-229640 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the Pil<sub>Mo</sub> pilin disulfide loop region (Pil<sub>Mo</sub>_DSL, also referred to as Pil<sub>20</sub>; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil<sub>20</sub> (SEQ ID NO: 60), Pil<sub>19</sub> (SEQ ID NO: 133), Pil<sub>18</sub> (SEQ ID NO: 134), Pil<sub>17</sub> (SEQ ID NO: 135), Pil<sub>16</sub> (SEQ ID NO: 136), Pil<sub>15</sub> (SEQ ID NO: 109), Pil<sub>14</sub> (SEQ ID NO: 137), Pil<sub>13</sub> (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil<sub>8</sub> (SEQ ID NO: 141), Pil<sub>7</sub> (SEQ ID NO: 113), Pil<sub>6</sub> (SEQ ID NO: 114), Pil<sub>5</sub> (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] Atty. Dkt. No.64100-229640 (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil<sub>18S[A]</sub> (SEQ ID NO: 185), Pil<sub>17S[A]</sub> (SEQ ID NO: 186), Pil<sub>16S[A]</sub> (SEQ ID NO: 187), Pil<sub>15S[A]</sub> (SEQ ID NO: 188), Pil<sub>14S[A]</sub> (SEQ ID NO: 189), Pil<sub>13S[A]</sub> (SEQ ID NO: 190), Pil<sub>12S[A]</sub> (SEQ ID NO: 191), Pil<sub>11S[A]</sub> (SEQ ID NO: 192), Pil<sub>10S[A]</sub> (SEQ ID NO: 193), Pil<sub>9S[A]</sub> (SEQ ID NO: 194), Pil<sub>8S[A]</sub> (SEQ ID NO: 195), Pil<sub>7S[A]</sub> (SEQ ID NO: 196), Pil<sub>6S[A]</sub> (SEQ ID NO: 197), Pil<sub>5S[A]</sub> (SEQ ID NO: 198), or Pil<sub>4S[A]</sub> (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 4. The glycoconjugate of any one of Paragraphs 1 to 3, wherein the acceptor protein is a fusion protein comprising a heterologous carrier protein and the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. 5. The glycoconjugate of any one of Paragraphs 1 to 4, wherein the acceptor protein is a fusion protein and the fusion protein further comprises an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus, optionally wherein the additional glycosylation sequence is an internal glycosylation fragment of ComP, further optionally, wherein the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13; Atty. Dkt. No.64100-229640 optionally, wherein the additional glycosylation sequence is also covalently linked to an oligo- or polysaccharide. 6. The glycoconjugate of Paragraph 5, wherein the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more additional glycosylation sequences; optionally, wherein the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five additional glycosylation sequences; optionally, wherein the additional glycosylation sequences are identical; optionally, wherein the additional glycosylation sequences differ from each other; and/or optionally, wherein at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other; and/or optionally, wherein none of the additional glycosylation sequences are the same. 7. The glycoconjugate of any one of Paragraphs 1 to 6, wherein the oligo- or polysaccharide covalently linked to the pilin-like protein or glycosylation fragment thereof has a size of at least three repeating units of oligo- or polysaccharide structure and/or has a size of at least ten monosaccharides. 8. The glycoconjugate of any one of Paragraphs 1 to 7, (i) wherein the oligo- or polysaccharide is produced by bacteria of the genus Streptococcus and the polysaccharide is capsular polysaccharide, optionally, wherein is S. pneumoniae or S. agalactiae, and optionally, wherein the S. agalactiae capsular polysaccharide is Ia, Ib, II, III, IV, V, VI, VII, VIII, or IX; (ii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Klebsiella and the polysaccharide is a capsular polysaccharide or O-antigen polysaccharide, optionally wherein the bacteria is K. pneumoniae; or (iii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Salmonella and the polysaccharide is O-antigen polysaccharide; optionally wherein the bacteria is S. enterica and the S. enterica polysaccharide is a Group B O-antigen. 9. The glycoconjugate of any one of Paragraphs 1 to 8, wherein the glycoconjugate is produced in vivo; Atty. Dkt. No.64100-229640 optionally, in a bacterial cell; optionally, in Escherichia coli; optionally, in a bacterium from the genus Klebsiella; and/or optionally, wherein the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 10. The glycoconjugate of any one of Paragraphs 1 to 9, wherein the bioconjugate is a conjugate vaccine that induces an immune response when administered to a subject; optionally, wherein the immune response elicits long term memory (memory B and T cells), is an antibody response, and is optionally a serotype-specific antibody response; optionally, wherein the antibody response is an IgG or IgM response; optionally, wherein the antibody response is an IgG response; optionally an IgG1 response; and/or optionally, wherein the conjugate vaccine generates immunological memory in a subject administered the vaccine. 11. A pilin-like-protein glycosylation fragment comprising or consisting of an isolated fragment of a TfpM-associated pilin-like protein, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) Pil<sub>Mo</sub> (SEQ ID NO: 57), (ii) Pil<sub>Mo</sub> lacking amino acids corresponding to residues 1–28 (Pil<sub>Mo</sub>g+1% E9C =8 @A3.1$% VY #PPP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine, (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 Atty. Dkt. No.64100-229640 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil₂₀ (SEQ ID NO: 60), Pil₁₉ (SEQ ID NO: 133), Pil₁₈ (SEQ ID NO: 134), Pil₁₇ (SEQ ID NO: 135), Pil₁₆ (SEQ ID NO: 136), Pil₁₅ (SEQ ID NO: 109), Pil₁₄ (SEQ ID NO: 137), Pil₁₃ (SEQ ID NO: 110), Pil₁₂ (SEQ ID NO: 138), Pil₁₁ (SEQ ID NO: 139), Pil₁₀ (SEQ ID NO: 112), Pil₉ (SEQ ID NO: 140), Pil₈ (SEQ ID NO: 141), Pil₇ (SEQ ID NO: 113), Pil₆ (SEQ ID NO: 114), Pil₅ (SEQ ID NO: 115), Pil₄ (SEQ ID NO: 116), or Pil₃ (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil_20[A] (SEQ ID NO: 166), Pil_19[A] (SEQ ID NO: 167), Pil_18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: Atty. Dkt. No.64100-229640 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil_8S[A] (SEQ ID NO: 195), Pil_7S[A] (SEQ ID NO: 196), Pil_6S[A] (SEQ ID NO: 197), Pil_5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 12. The pilin-like-protein glycosylation fragment of Paragraph 11, wherein the pilin-like- protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 13. A fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof translationally fused to a heterologous carrier protein, wherein the TfpM-associated pilin-like protein or glycosylation fragment comprises a C- terminus serine or threonine residue, wherein the TfpM-associated pilin-like protein or glycosylation fragment is the C- terminus-most sequence of the fusion protein, and wherein the fusion protein comprises a C-terminus serine or threonine residue; Atty. Dkt. No.64100-229640 optionally, wherein the fusion protein is glycosylated by an olio- or polysaccharide covalently linked to the C-terminus serine or threonine; optionally, wherein the fusion protein is glycosylated by an oligo- or polysaccharide comprising glucose at its reducing end covalently linked to the C-terminus serine or threonine. 14. The fusion protein of Paragraph 13, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 15. The fusion protein of Paragraph 13 or 14, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) Pil_Mo (SEQ ID NO: 57), (ii) Pil_Mo lacking amino acids corresponding to residues 1–28 (Pil_Mog+1% E9C =8 @A3.1$% VY #PP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of Pil<sub>DSM16617</sub> (SEQ ID NO: 82), Pil<sub>ZZC3-9</sub> (SEQ ID NO: 83), Pil<sub>TUM15069</sub> (SEQ ID NO: 84), Pil<sub>AI7</sub> (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Pil<sub>AI40</sub> (SEQ ID NO: 89), Pil<sub>F78</sub> (SEQ ID NO: 90), Pil<sub>S71</sub> (SEQ ID NO: 91), Pil<sub>ANC4282</sub> (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: Atty. Dkt. No.64100-229640 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil<sub>19</sub> (SEQ ID NO: 133), Pil<sub>18</sub> (SEQ ID NO: 134), Pil<sub>17</sub> (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil<sub>12</sub> (SEQ ID NO: 138), Pil<sub>11</sub> (SEQ ID NO: 139), Pil<sub>10</sub> (SEQ ID NO: 112), Pil<sub>9</sub> (SEQ ID NO: 140), Pil<sub>8</sub> (SEQ ID NO: 141), Pil<sub>7</sub> (SEQ ID NO: 113), Pil<sub>6</sub> (SEQ ID NO: 114), Pil<sub>5</sub> (SEQ ID NO: 115), Pil<sub>4</sub> (SEQ ID NO: 116), or Pil<sub>3</sub> (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil<sub>20[A]</sub> (SEQ ID NO: 166), Pil<sub>19[A]</sub> (SEQ ID NO: 167), Pil<sub>18[A]</sub> (SEQ ID NO: 168), Pil<sub>17[A]</sub> (SEQ ID NO: 169), Pil<sub>16[A]</sub> (SEQ ID NO: 170), Pil<sub>15[A]</sub> (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil<sub>10[A]</sub> (SEQ ID NO: 176), Pil<sub>9[A]</sub> (SEQ ID NO: 177), Pil<sub>8[A]</sub> (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Atty. Dkt. No.64100-229640 Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 16. The fusion protein of any one of Paragraphs 13 to 15, wherein the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. 17. The fusion protein of any one of Paragraphs 13 to 16, wherein the fusion protein further comprises an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus, optionally wherein the additional glycosylation sequence is an internal glycosylation fragment of ComP, further optionally, wherein the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13; optionally, wherein the additional glycosylation sequence is also covalently linked to an oligo- or polysaccharide. 18. The fusion protein of Paragraph 17, wherein the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more additional glycosylation sequence; optionally, wherein the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five additional glycosylation sequence; optionally, wherein the additional glycosylation sequence are identical; Atty. Dkt. No.64100-229640 optionally, wherein the additional glycosylation sequence differ from each other; and/or optionally, wherein at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other; and/or optionally, wherein none of the additional glycosylation sequence are the same. 19. The fusion protein of any one of Paragraphs 13 to 18, wherein the oligo- or polysaccharide covalently linked to the pilin-like protein or glycosylation fragment thereof has a size of at least three repeating units of oligo- or polysaccharide structure and/or has a size of at least ten monosaccharides. 20. The fusion protein of any one of Paragraphs 13 to 19, (i) wherein the oligo- or polysaccharide is produced by bacteria of the genus Streptococcus and the polysaccharide is capsular polysaccharide, optionally, wherein is S. pneumoniae or S. agalactiae, and optionally, wherein the S. agalactiae capsular polysaccharide is Ia, Ib, II, III, IV, V, VI, VII, VIII, or IX; (ii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Klebsiella and the polysaccharide is a capsular polysaccharide or O-antigen polysaccharide, optionally wherein the bacteria is K. pneumoniae; or (iii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Salmonella and the polysaccharide is O-antigen polysaccharide; optionally wherein the bacteria is S. enterica and the S. enterica polysaccharide is a Group B O-antigen. 21. The fusion protein of any one of Paragraphs 13 to 20, wherein the glycosylated fusion protein is produced in vivo; optionally, in a bacterial cell; optionally, in Escherichia coli; optionally, in a bacterium from the genus Klebsiella; and/or optionally, wherein the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 22. The fusion protein of any one of Paragraphs 13 to 21, wherein when the fusion protein is a vaccine that induces an immune response when administered to a subject; optionally, wherein the immune response elicits long term memory (memory B and T cells), is an antibody response, and is optionally a serotype-specific antibody response; Atty. Dkt. No.64100-229640 optionally, wherein the antibody response is an IgG or IgM response; optionally, wherein the antibody response is an IgG response; optionally an IgG1 response; and/or optionally, wherein the fusion protein generates immunological memory in a subject administered the vaccine. 23. A method of producing a glycoconjugate, the method comprising covalently linking an oligo- or polysaccharide to an acceptor protein comprising or consisting of a TfpM-associated pilin-like protein or glycosylation fragment thereof using a TfpM oligosaccharyltransferase (OTase); wherein the pilin-like protein or glycosylation fragment comprises a C-terminus serine or threonine residue, the acceptor protein comprises a C-terminus serine or threonine residue, and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine residue of the acceptor protein; optionally, wherein the oligo- or polysaccharide comprises a glucose at its reducing end; optionally, wherein the acceptor protein is a fusion protein of any one of Paragraphs 13 to 22; optionally, wherein the method is a method of in vivo conjugation of an oligo- or polysaccharide to an acceptor protein; and/or optionally, wherein the glycoconjugate is immunogenic. 24. The method of Paragraph 23, wherein the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpM<sub>Mo</sub> (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpM<sub>AI7</sub> (SEQ ID NO: 66), TfpM<sub>VE-C3</sub> (SEQ ID NO: 67), TfpM<sub>YH01026</sub> (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 Atty. Dkt. No.64100-229640 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), or TfpM72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpMMo (SEQ ID NO: 56). 25. The method of Paragraph 23 or 24, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 26. The method of any one of Paragraphs 23 to 25, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) Pil<sub>Mo</sub> (SEQ ID NO: 57), (ii) Pil<sub>Mo</sub> lacking amino acids corresponding to residues 1–28 (Pil<sub>Mo</sub>g+1% E9C =8 @A3.1$% VY #PP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), Pil_TUM15069 (SEQ ID NO: 84), Pil_AI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Pil_AI40 (SEQ ID NO: 89), Pil_F78 (SEQ ID NO: 90), Pil_S71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: Atty. Dkt. No.64100-229640 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil₁₉ (SEQ ID NO: 133), Pil₁₈ (SEQ ID NO: 134), Pil₁₇ (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil₁₂ (SEQ ID NO: 138), Pil₁₁ (SEQ ID NO: 139), Pil₁₀ (SEQ ID NO: 112), Pil₉ (SEQ ID NO: 140), Pil₈ (SEQ ID NO: 141), Pil₇ (SEQ ID NO: 113), Pil₆ (SEQ ID NO: 114), Pil₅ (SEQ ID NO: 115), Pil₄ (SEQ ID NO: 116), or Pil₃ (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil_20[A] (SEQ ID NO: 166), Pil_19[A] (SEQ ID NO: 167), Pil_18[A] (SEQ ID NO: 168), Pil_17[A] (SEQ ID NO: 169), Pil_16[A] (SEQ ID NO: 170), Pil_15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil_10[A] (SEQ ID NO: 176), Pil_9[A] (SEQ ID NO: 177), Pil_8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Atty. Dkt. No.64100-229640 Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 27. The method of any one of Paragraphs 23 to 26, wherein the acceptor protein is a fusion protein comprising a heterologous carrier protein and the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. 28. The method of any one of Paragraphs 23 to 27, wherein the acceptor protein is a fusion protein comprising an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus, and the method further comprises covalently linking an oligo- or polysaccharide to the additional glycosylation sequence with an OTase other than TfpM OTase; optionally, wherein the acceptor protein is a fusion protein comprising a ComP glycosylation fragment, and the method further comprises covalently linking an oligo- or polysaccharide to the ComP glycosylation fragment using a PglS OTase, optionally, wherein the ComP glycosylation fragment is an internal glycosylation fragment of ComP, further optionally, wherein the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13. 29. The method of any one of Paragraphs23 to 28, wherein the conjugation occurs in a host cell. Atty. Dkt. No.64100-229640 30. The method of Paragraph 29, wherein the host cell is a bacterial cell; optionally, in Escherichia coli; optionally, in a bacterium from the genus Klebsiella; or optionally, wherein the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 31. The method of Paragraph 29 or 30, comprising culturing a host cell that comprises: (a) a genetic cluster encoding for the proteins required to synthesize the oligo- or polysaccharide; (b) a TfpM OTase; and (3) the acceptor protein. 32. The method of any one of Paragraphs 23 to 31, wherein the method produces a conjugate vaccine. 33. A host cell comprising (a) a genetic cluster encoding for the proteins required to synthesize an oligo- or polysaccharide; (b) a TfpM OTase; and (3) an acceptor protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof. 34. The host cell of Paragraph 33, wherein the acceptor protein is a fusion protein. 35. The host cell of Paragraph 33 or 34, wherein the host cell comprises a nucleic acid encoding the TfpM OTase; and/or wherein the host cell comprises a nucleic acid encoding the acceptor protein; optionally, wherein the TfpM OTase and the acceptor protein are encoded by the same nucleic acid. 36. An isolated nucleic acid encoding the pilin-like-protein glycosylation fragment of Paragraph 11 or 12 and/or the fusion protein of any one of Paragraphs 13 to 22. 37. The isolated nucleic acid of Paragraph 36, wherein the nucleic acid is a vector. 38. A host cell comprising the isolated nucleic acid of Paragraph 36 or 37, optionally, wherein the host cell is a bacterial cell; and further: optionally, wherein the host cell is Escherichia coli; optionally, wherein the host cell is from the genus Klebsiella; or optionally, wherein the host cell is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 39. A composition comprising the conjugate vaccine of Paragraph 10 or the fusion protein of Paragraph 22, and an adjuvant and/or carrier. 40. A method of inducing a host immune response against a bacterial pathogen, the method comprising administering to a subject in need of the immune response an effective amount of the Atty. Dkt. No.64100-229640 conjugate vaccine of Paragraph 10, the fusion protein of Paragraph 22, or the composition of Paragraph 39. 41. The method of Paragraph 40, wherein the immune response is an antibody response; wherein the immune response is selected from the group consisting of an innate response, an adaptive response, a humoral response, an antibody response, cell mediated response, a B cell response, a T cell response, cytokine upregulation or downregulation, immune system cross-talk, and a combination of two or more of said immune responses; and/or wherein the immune response is selected from the group consisting of an innate response, a humoral response, an antibody response, a T cell response, and a combination of two or more of said immune responses. 42. A method of preventing or treating a bacterial disease and/or infection in a subject comprising administering to a subject in need thereof the conjugate vaccine of Paragraph 10, the fusion protein of Paragraph 22, or the composition of Paragraph 39; optionally, wherein the subject is a human. 43. The method of Paragraph 42, wherein the infection is a localized or systemic infection of skin, soft tissue, blood, or an organ, or is auto-immune in nature; wherein the disease is pneumonia; and/or wherein the infection is a systemic infection and/or an infection of the blood. 44. The method of any one of Paragraphs 40 to 43, wherein the conjugate vaccine, the fusion protein, or the composition is administered via intramuscular injection, intradermal injection, intraperitoneal injection, subcutaneous injection, intravenous injection, oral administration, mucosal administration, intranasal administration, or pulmonary administration. 45. A method of producing a pneumococcal conjugate vaccine against pneumococcal infection, the method comprising: (a) isolating the glycoconjugate of any one of Paragraphs 1 to 10 or a glycosylated fusion protein of any one of Paragraphs 13 to 22; and (b) combining the isolated glycoconjugate or isolated glycosylated fusion protein with an adjuvant and/or carrier. Atty. Dkt. No.64100-229640 46. The glycoconjugate, glycosylated fusion protein, or conjugate vaccine of any of the above claims for use in inducing a host immune response against a bacterial pathogen and/or preventing or treating a bacterial disease and/or infection in a subject. 47. A recombinant nucleic acid construct comprising a nucleotide sequence encoding a TfpM oligosaccharyltransferase (OTase) operably linked to at least one heterologous transcriptional regulatory sequence. 48. The recombinant construct of Paragraph 47, wherein the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpM_ANC4282 (SEQ ID NO: 73), TfpM_CIP102159 (SEQ ID NO: 74), TfpM_junii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpM_BI730 (SEQ ID NO: 79), TfpM_A3K91 (SEQ ID NO: 80), and/or TfpM_72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpM_Mo (SEQ ID NO: 56), TfpM_DSM16617 (SEQ ID NO: 63), TfpM_ZZC3 (SEQ ID NO: 64), TfpM_TUM15069 (SEQ ID NO: 65), TfpM_AI7 (SEQ ID NO: 66), TfpM_VE-C3 (SEQ ID NO: 67), TfpM_YH01026 (SEQ ID NO: 68), TfpM_CIP102143 (SEQ ID NO: 69), TfpM_AI40 (SEQ ID NO: 70), TfpM_F78 (SEQ ID NO: 71), TfpM_S71 (SEQ ID NO: 72), TfpM_ANC4282 (SEQ ID NO: 73), TfpM_CIP102159 (SEQ ID NO: 74), TfpM_junii-65 (SEQ ID NO: 75), TfpM_YZS-X (SEQ ID NO: 76), TfpM_CIP102637 (SEQ ID NO: 77), TfpM_T-3-2 (SEQ ID NO: 78), TfpM_BI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpM_Mo (SEQ ID NO: 56). 49. The recombinant construct of Paragraph 47 or 48, wherein the heterologous transcriptional regulatory sequence is a promotor sequence. 50. The recombinant construct of any one of Paragraphs 47 to 49, further comprising a nucleotide sequence encoding a TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof operably linked to the nucleotide sequence encoding the a TfpM OTase; optionally, wherein the coding sequence of the TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like Atty. Dkt. No.64100-229640 protein or glycosylation fragment thereof is within 2, 5, 10, 20, 30, 40, or 50 nucleotides of the sequence encoding the TfpM OTase; optionally, wherein the coding sequence of the TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof overlaps the operably linked nucleotide sequence encoding the a TfpM OTase. 51. The recombinant construct of Paragraph 50, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 52. The recombinant construct of Paragraph 50 or 51, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) Pil_Mo (SEQ ID NO: 57), (ii) Pil_Mo lacking amino acids corresponding to residues 1–28 (Pil_Mog+1% E9C =8 @A3.1$% VY #PP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ Atty. Dkt. No.64100-229640 ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C-terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil<sub>19</sub> (SEQ ID NO: 133), Pil<sub>18</sub> (SEQ ID NO: 134), Pil<sub>17</sub> (SEQ ID NO: 135), Pil<sub>16</sub> (SEQ ID NO: 136), Pil<sub>15</sub> (SEQ ID NO: 109), Pil<sub>14</sub> (SEQ ID NO: 137), Pil<sub>13</sub> (SEQ ID NO: 110), Pil<sub>12</sub> (SEQ ID NO: 138), Pil<sub>11</sub> (SEQ ID NO: 139), Pil<sub>10</sub> (SEQ ID NO: 112), Pil<sub>9</sub> (SEQ ID NO: 140), Pil<sub>8</sub> (SEQ ID NO: 141), Pil<sub>7</sub> (SEQ ID NO: 113), Pil<sub>6</sub> (SEQ ID NO: 114), Pil<sub>5</sub> (SEQ ID NO: 115), Pil<sub>4</sub> (SEQ ID NO: 116), or Pil<sub>3</sub> (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil<sub>20[A]</sub> (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil<sub>16[A]</sub> (SEQ ID NO: 170), Pil<sub>15[A]</sub> (SEQ ID NO: 171), Pil<sub>14[A]</sub> (SEQ ID NO: 172), Pil<sub>13[A]</sub> (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C- terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Atty. Dkt. No.64100-229640 Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 53. The recombinant construct of any one of Paragraphs 50 to 52, wherein the fusion protein is a fusion protein of any one of Paragraphs 13 to 22. 54. The recombinant construct of any one of Paragraphs 50 to 53, further comprising a nucleotide sequence encoding a PglS OTase operably linked to the TpfM OTase; optionally, wherein the coding sequence of the PglS OTase is within 10, 20, 30, 40, 50, 75, or 100 nucleotides of the sequence encoding the TfpM OTase. 55. A vector comprising the recombinant nucleic acid construct of any one of Paragraphs 47 to 54. 56. A host cell comprising the recombinant nucleic acid construct of any one of Paragraphs 47 to 54, or the vector of Paragraph 55; optionally, wherein the host cell is a bacterial cell; and further: optionally, wherein the host cell is Escherichia coli; optionally, wherein the host cell is from the genus Klebsiella; or optionally, wherein the host cell is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 57. A method for producing a TfpM OTase, the method comprising culturing the host cell of Paragraph 56, wherein said vector of Paragraph 54 is an expression vector, and recovering the TfpM OTase. Atty. Dkt. No.64100-229640 58. A glycoconjugate comprising an oligo- or polysaccharide covalently linked to an acceptor protein, wherein the acceptor protein comprises means for being covalently linked to the oligo- or polysaccharide by a TfpM OTase. 59. The glycoconjugate of Paragraph 58, wherein the glycoconjugate is the glycoconjugate of any one of Paragraphs 1 to 10. 60. A fusion protein comprising means for being covalently linked to an oligo- or polysaccharide by a TfpM OTase, wherein said means is translationally fused to a heterologous carrier protein, wherein the means is located at the C-terminus-most sequence of the fusion protein. 61. The fusion protein of Paragraph 60, wherein the fusion protein is the fusion protein of any one of Paragraphs 13 to 22. 62. The glycoconjugate of Paragraph 58 or 59 or the fusion protein of Paragraph 60 or 61, for use in any of the methods above. Atty. Dkt. No.64100-229640 ADDITIONAL SEQUENCES >TfpM_Mo (SEQ ID NO: 56) MFNLIKHFKNKELFVALYFLLFIVGFSIYSFNFYNELRVLEVFLLIGLGLYGLIHKKNYFSKYEFCF LIFTLLGFLFWSNYSYIFYELILFYLLYKAFFNLNYNEFLSKLLIWLSFSIFLMLPIAITEFILSGVYPN WYPMPWNIRVYNSYFLVLSIFAVWFYLKQEKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIACV CIFNHAARFKLLFTYVATWLAYFSILFFSSHSASIVGSIVRESTSGRYELWVSAFQCWSQNPIFGCG FYQLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSFS GIHIYPVTQVALLWLFVFLLKNPEFQHAASFHPSTYEASTVSQVFGYIAYISIALAFIYLFFYTSALS ESLPSTPPRFWEYGYQLF >TfpM_DSM16617 (SEQ ID NO: 63) MLNFNKHLKNKELFIALYFLTFILGLSIHISSNFYNESRVLETFLLLSLVFNKNLHLKKIEYIFIIFIIF HLLFLRNSQFIIFEILLYYLLYKAFLIINYNELLAKFIIWISFSIFSILPIEIVKYIHNSIYSNWYPTPWNI RVYNSYFLVLSVFAVWFYLKQEKFKIIYLFFLFLAFLSILLDGGRSATLAYSVFIACVCIFNRAVRF KLLFTYAATWLTYFSILFFSSHSASNVRSIVRESTSGRYELWVSAFQCWSQNPIFGCGFYQLDQYS GVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSFSGIHIYPVTQ VALLWLFVFLLKNPEFEHAASFHPSTYEASRVSQVFGYIAYISIALAFIYLFFYTSALSESLASTPPR FWEYGYQLF >TfpM_ZZC3 (SEQ ID NO: 64) MLNFNKYLKNKELFIALYFLSFILGLSIHISSNFYNESRVLETFLLLSLVFNKNLHLKKIEYIFIIFIIF HLFFLRNSQFIIFEILLYYLLYKAFLVINYNELLAKFIIWISFSIFSMLPIEIVKYIHNSIYSNWYPTPW NIRVYNSYFFVLSIFAIWFYLKQEKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIACVCIFNRAAR FKLLFTYAATWLTYFSILFFSSHSASNVRSIVRESTSGRYELWVSAFQCWSQNPIFGCGFYQLDQY SGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSFSGIHIYPVT QVALLWLFVFLLKNPEFQHAASFHPSTYEASRVSQVFGYIAYISIALAFIYLFFYTSALSESLPSTPP RFWEYGYQLF >TfpM_TUM15069 (SEQ ID NO: 65) MFNLIKHFKNKELFVALYFFLFILGVSINGFNFYNELRVVEVFLLIFLGFYGLINKKNYFSKYEVCF LIFTLLGFLFWSNYSYIFYELILFYLLYKAFLNLNYNEFLSKFVIWLSFLIFLMLPIAITEFIASGVYQ NWYPMPWNIRVYNSYFLVLSIFAVWFYLKQEKFKNIYLVFLFLAFLSILLDGGRSATLAYSVFIAC VCIFNHAARFKLLFTYVATWLAYFSILFFSSHSASIVGSIVRESTSGRYELWVSAFQCWSQNPIFGC GFYQLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLS FSGIHIYPVTQVALLWLFVFLLKNPEFEHAASFHPSTYEASRVSQVFGYIAYISIALAFIYLFFYTSA LSESLASTPPRFWEYGYQLF >TfpM_AI7 (SEQ ID NO: 66) MFKYIKHFKNKELFIALYFLLFIVGFTIYSFNFYNELRAIEFFLLIFFGFHSLINKNYVSKYEFCFLIFI FLGFLFWSNYSYIFYELILFYLLYKAFLNLNYNEFLSKFVIWLSFLIFLMLPIAITEFILSGVYQNWY PMPWNIRVYNSYFLVLSIFAVWFYLKQEKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIACVCMF NRAVRFKLLFTYAATWLVYFSILFISSHSASGVRSIVRESTSGRYELWVNAFQCWSQNPIFGCGFY QLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSFSGI HIYPVTQVALLWLFVFLLKNPEFEHAASFHPSTYEASRVSQVFGYIAYILIALAFIYLFSYTSALSES LPSTPPRFWEYGYQLF >TfpM_VE-C3 (SEQ ID NO: 67) MNFLVHFKNKELFVALYFLLFTVGFSIYSFNFYNELRALEFFLLIGLGLYGLIHKKNYFSKYEFCFL IFTLLGFLFWSNYYYIFFELILFYLLYKAFLNLNYNEFLSKLLIWLSFSIFLMLPIAITEFILSGVYQN WYPMPWNIRVYNSYFLVLSIFAVWFYLKQEKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIACV CMFNRAARFKLLFTSAATWLTYFSILFFSSHSASNVRSIVRESTSGRYELWVSAFQCWSQNPIFGC Atty. Dkt. No.64100-229640 GFYQLDHYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLS FSGIHIYPVTQVALLWLFVFLLKNPEFQHAASFHPSTYEASRVSQVFGYIAYISIALAFIYLFFYTSA LSESLPSTPPRFWEYGYQLF >TfpM_YH01026 (SEQ ID NO: 68) MFNLIKHFKNKELFIALYFLLFIVGFSIYSFNFYNELRVLEVFIFIGLVLFGLIHKKNYFSKYEFCFLI FTLLGFLFWSNYSYIFYELILFYLLYKAFLNLNYNEFISKLVIWLSFSIFLMLPIAITEFMLSGLYQN WYPMPWNIRVYNSYFLVLSIFAVWFYFKQEKFINIYLVFLFLAFLSILLDGGRSATLAYSVFIACV CIFNRAARFKLLFIYVATWLTYFSILFFSSHSASNVRSIVRESTSGRYELWVSAFQCWSQNPIFGCG FYQLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVICNILKRIKWNLKENYFVISAFIAVSIDLSFS GIHIYPITQVALLWLFVFLLKNPEFQHAASFHPSTYEASRVSQVFGYIAYILIALAFIYLFFYTSALS ESLPSTPPRFWEYGYQLF >TfpM_CIP102143 (SEQ ID NO: 69) MFNLIKHLKNKELFVALYFLLFIVGFSIYSSNFYNELRALEFFLLIGLGLYVLIHKKNYFSKYELCF LIFTLLGFLFWSNYSYIFYELILFYLLYKAFLNLNYNEFLSKLLIWLSFSIFLMLPIAITEFIISGVYQN WYPMPWNIRVYNSYFLVLSIFAVWFYLKQEKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIACV CIFNHAARFKLLFTYAVTWLAYFSILFFSSHSTSIVGSIVRESTSGRYELWVSAFQCWSQNPIFGCG FYQLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSFS GIHIYPITQVALLWLFVFLLKNPEFQHATSFHPPIYEASKVSQVFEFMVYILIALAFIYLFFYTSALS ESLASTPPRFWEYGYQLF >TfpM_AI40 (SEQ ID NO: 70) MFSVVSHLKNKELFIALYFLLFISGISIYSYNFYNELRVSEVFLLIGLGLCGLTHKKNYFSKYEFYFF IFILLGFLFWSNYNYILYEFILFYLLYQAFLNLNYSEFLSKFVIWLSFSIFLMFPIAVAEFLVSGVYQ NWYPMPWNIRIYNSYFLVFSIFSVWFYLKEEKFKNIYLFFLFLAFLSILLDGGRSATLAYSIFIACIC MFNRTARLKILFTYMATWLTYFSILFFSSNSASGVRSIARESTSGRYELWVSAFQCWLQNPLFGC GFYQLDLYSGIIPAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSF SGIHIYPVTQVALLWLFVFLLKNPEFQHAASFHSSTYEASRVSQVFGYIVCILIALAFIYLFFYTSAL SESLPSTPPRFWEYGYQLF >TfpM_F78 (SEQ ID NO: 71) MSNLIKYLKNKELFIALYFFIFILGISIQLSFNFYNEARVLEILLLLCLGLYSFINNENLFFKKELVFLF FISFGFFYWFNFQIVFYEILLFYLLYKAFFFLKYNAIVSKLIVFSSFFIFIFLPMSLWEYLTTGKYQN WYPLPWNIRIYNSYFLIFSIFAIWFFLKEKYKSIYLAFIFLAFLSILLDGGRSAALAYTVFIGLLSIFN RLARLKLIFIYSLTWLAYFLIIYFSSQSGSSLRYIARDSTSGRYDLWLNAFQCWLQSPILGCGFYQL DKYSNLSAHPHNLFIQILTETGLIGLSFLLYIIFIILRNISWKFKENYFVISALIAVFIDLSFSGIHIYPIT QVALLWLFVFLLKNPEFQHATYFSPSACEASKKSQFLEFIVYILIALAFIYLFFNTSALSESLPSTPP RFWEYGYQIF >TfpM_S71 (SEQ ID NO: 72) MSNLIRHLKNKELFLALYFLTFILGVSLGVSYNFYNEARVLEVLLLLGFGFYSVFSKEIFFSKKEYL FLVVFISYLFFLKNSQFIIFDILLFYLLYKSFFILNYNLVVSKIIVLSSFLIFMTFPLSLLGYWGDGVY RNWYPMPWNIRVYNSYFLILLIFSTWLLMRGNRYTWVYLLFTYLSLLSILLDGGRSALLAYSTFFI IVIIFNKKVRLKLIFIYIISWLSFLLIVFSAGIASDGISIARVTTSRRSDLWMHALQCWIESPIFGCGFY QLGAYENLSAHPHNLFIQILTETGVMGFSFLALIIFGVLRNISWNIKENYFVIAAFFAIGVDLSFSGI HIYPITQVGLLWLFVFLLKNPEFRHAKYFSDILVQNPKSVWVVNFIIYLIITCAFIYLFVNTSALSES LAVTPPRFWEYGYQLF >TfpM_ANC4282 (SEQ ID NO: 73) MLNLIKNKELFIALYFLIFNLGFSVHISSNFYNEARLLEIFLLLSLGIFSGFVKNIVFHKIEYIFLLFFI FSIFFLKNQPFIFFEILLFYLLFKAFFALNYNSKISKAIILLSFLIFLMFPVSILHYLNSGLYQNWYPMP WNIRIYNSYFLIFSIFAIWFYLKEDKYKNIYLIFIFLAFLSILLDGGRSATLAYTIFIVIVCIFNRLERF Atty. Dkt. No.64100-229640 KLLLIYCSTWLAYFSIVYFSSQSASTLRSITRESTSGRYELWLNAFQCWLENPILGCGFYQLDKYPS LSAHPHNLFVQILTETGLIGFIFLSFIIFKVVKNISWNFKQNYFVLAALFSVAIELSFSGIHIYPVTQV ALLWLFVFLLKNPEFSHASYFNYLKIKNSKIDKLIQLFIYLFILIIFIYLFINTSVLSENLPSTPPRFWE YGYQLF >TfpM_CIP102519 (SEQ ID NO: 74) MFNLIKHIKNKELFLALYFLLFSVGFSIYSFNFYNELRALEFFLLIGLGLYGLIHKKNYFSKYEVCF LIFTFLGFSFWSNYSYIFYELILFYLLYKAFLNLNYNEFLSKLVIWLSFSIFLMLPIAITEFMLNGIYQ NWYPMPWNIRVYNSYFLVLSIFAVWFYLKQDKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIAC VCMFNRAARFKLLFTYAVTWLVYFSILFFSSHSDSNVSSIVRESTSGRYELWVSAFQCWSQNPIFG CGFYQLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDL SFSGIHIYPITQVALLWLFVFLLKNPEFEHAASFYPSTYEASRVSQVFGYIAYILIALAFIYLFFYTSA LSESLPSTPPRFWEYGYQLF >TfpM_junii_65 (SEQ ID NO: 75) MNFLVHFKNKELFVALYFLLFSVGFSIYSFNFYNELRVLEIFLFIGLGLNSLTHKKNYFSKYEFCFL IFTLLGFLFWSNYSYIFFELILFYLLYKAFLNLNYNEFLSKLVIWLSFSIFLMLPIAIKEFIASGVYQN WYPMPWNIRVYNSYFFILSIFAIWFYSKQEKFKNIYLIFLFLAFLSILLDGGRSATLAYSVFIACVC MFNRAARFKLLFTYAVTWLAYFSILFFSSHSTSIVGSIVRESTSGRYELWVSAFQCWSQNPIFGCG FYQLEQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLSFS GIHIYPVTQVALLWLFVFLLKNPEFQHAASFHPSTYEASRVSQVFGYIVYILIALAFIYLFSYTSALS ESLPSTPPRFWEYGYQLF >TpfM_YZS-X (SEQ ID NO: 76) MFNLIKHFKNKELFIALYFLLFIAGFSIDSFNFYNELRIFEVFLLIGLGFYGLTHKKKYFSKYEFCFFI FILLGFLCWSNYSYILYEFILFYLLYKAFFDLNYNEFLSKLIIWLSFSIFLMFPIAVAEFLVSGVYQN WYPMPWNIRIYNSYFLVFSIFSVWFYLKEEKFKNIYLFFLFLAFLSILLDGGRSANLAYSIFIACICM FNRTARLKILFTYIATWLTYFSILFFSSNSASGVRSIARESTSGRYELWVSAFQCWLQNPLFGCGFY QLDLYSGIIPAHPHNLFIQILTETGLIGFGFLILVIWNILKRIKWNFKENYFVIPALIAVSIDLSFSGIHI YPITQIALLWLFVFLLKNPEFQYAASFNQSTHKGSKKSKNFELIVYILIALAFIYLFFHTSALSESLPS TPPRFWEYGYRLF >TfpM_CIP102637 (SEQ ID NO: 77) MSNLIKHLKNKELLVALYFFLFILGVSINGFNFYNELRTLEVFLLIGLGFYSLIQKKSYFSKYEFCFL IFTLLGFLFWSNYSYIFYELILFYLLYKAFLNLNYNEFLSKLVIWLSFSIFLMLPIAITEFMLSGVYQ NWYPMPWNIRVYNSYFLVLSIFAIWFYLKQEKFNNIYLIFLFLAFLSILLDGGRSTALAYSVFIACV CMFNRAVRFKLLFTYAATWLTYFSILFFSSHSTSSVRSIVRESTSGRYELWVSAFQCWSQNPIFGC GFYQLDQYSGVISAHPHNLFIQVLTETGLIGFGFLILVIWNILKRIKWNLKENYFVISAFIAVSIDLS FSGIHIYPVTQVALLWLFVFLLKNPEFQRAASFHPSTYEASRVSQVFGYIAYILIALAFIYLFFYTSA LSESLPSTPPRFWEYGYQLF >TfpM_T-3-2 (SEQ ID NO: 78) MMKAIKLLAHKESFIGLYFLLFIMGVTVGSGYGIYNESRIAEIALLLGLGAHACFNKYYIVTKVEY LFFVFIIIGSFFWSNSFFIIIDLLLVYLLYKSFFFLEYRPLLTKIIVLASFLIFLLLPVAIWDYITSGIYTS NWYLLRLNIRIYNSYFLIMSIFAVWLYLTEKNYKKLYLSFIFLAFLSILMDGGRSATLAYTAFVIIIC IFRRPVSWQLGFAYSMSWLTYLTINHLASLNAVETLGLGIARATTSQRYDIWMNAVQCWVQNPI WGCGFYQLDSTRNLASHPHNLFLQVLSETGLIGFGFLLAIIFSILKNISWNLNKDYFVIAALLAVVI ETLLSGIHIYPITQIALLWLFIFLLKNPIFPHTLYFNSLIASFASNSYLSITVYLILTIFFLYFFINTSALI DPELLTRPRFLENGYNIF >TfpM_BI730 (SEQ ID NO: 79) MSTIVKRLKNKELFIALYFLLFILGFTIGITPKFYNEFRILQVTLLLNFGLHNIIHKHGYISRAELLFF VYIGIASLFWQNYEFIVIDLLLAYLLYKTFFLLKYNELATKVIVFFSLLIFPLLPLSVFDYISTGTYYP Atty. Dkt. No.64100-229640 IWHPMPWNIRIYDSYLLIVSIFAVWFYITETKYKKIYLLFLLLAFFSVLLNGGRSATLAYTVFIAVIV VFNRIVRWQILATYAIAWLAYISISYLAISNLSMASPIGLQIARTTTSLRYDLWMNAIECWIQSPLV GCGFYQLYRYENLGAHPHNLLLQILTETGLIGFGFLLAIVVTILKHIDWQLKRSYFVIAALLAIGV DTSLSGTHIYPITQMALLWLLVFLLKNPVFQHAAYFNRVPYITSVTDMVVSIVVYFSLTVIFIYLFL NTSVLFDSLMLATPPRFWEYGYQLF >TfpM_A3K91 (SEQ ID NO: 80) MYKKQLLNNQQMTVSIIKALANKESFVALYSLIFIVGVTANSGYGIYNESRIFEVALLLGLGAHAC FNKYYIVTKVEYLFFVFIIIGSFFWSNSFFIIIDLLLVYLLYKSFFFLEYRPLLTKIIVLASFLIFLLLPV AIWDYITSGIYTSNWYLLRLNIRIYNSYFLIMSIFAVWLYLTEKNYKKLYLSFIFLAFLSILMDGGR SATLAYTAFVIIICIFRRPVSWQLGFAYSMSWLTYLTINHLASLNAVETLDLGIARATTSQRYDIW MNAVQCWVQNPIWGCGFYQLDSTRNLAAHPHNLFLQVLSETGLIGFWFLLAIIFFILKNISWNLN KDYFVIASLLAVVIETLLSGIHIYPITQIALLWLFVFLLKSSNFSHAIYFHSSIPSFVVNRYFSVTMFL VLVILFIYFFINTSALLDNEIMTRPRFLENGYNIF >TfpM_72-O-c (SEQ ID NO: 81) MYNKNPLSKQHKLQTFIKSVRLLFNKELSIGLYFLLFILGMNTGSGHDSYNEFRVFQVTLLLVIGV STWCYRRLFITKLELLFFAFIAFGSFFWQQPIFVLNDVLLVYLLYKSFYLLNYQPLLSKLIVLSSLLI FLLLPVALWNYIDTGKYSPIWYPLPWNMRVYDSYFLIISVFAVWFYLTEKQYRFLYLLFLFLAFL AVLLDGGRSVTLAYTVFIAIISLFHRRARWRLVLMYAMSWLTYIIVTYTANTSVTSLRIARDTRSD RYDLWINAFQCWSQHPLFGCGFYQLDKYPNIAAHPHNLFIQVLTETGLIGFGFLAFIIFKVAKNIN WDLKQNYFVIAALLAISIDMSLSGVHIYPVTQIALLWLFVFLLKNPEFSHAHHFNKVVQQKKAID NILPLIIYLSLTIWFIYLFTNTSSFLPGTPLTPPRFWVYGYQLW >Pil_DSM16617 (SEQ ID NO: 82) MNTMQKGFTLIELMIVVAIIGILAAIAIPAYQDYTVRARVSEGITTASAMKATVSENILNAGALVA GTPSTAGSACVGVTEISGGTGNVASATCGASKAGQIIVTMGATTAKSVPVYLTPSYTASAVTWSC STTAGNEKYVPSECRKVGT >Pil_ZZC3-9 (SEQ ID NO: 83) MNAQKGFTLIELMIVVAIIGILAAIAIPAYQDYTVRARVSEGITTASAMKATVSENILNAGALVAG APSTAGSACVGVTEISGGTGNVASATCGASKPGQIIVTMGATTAKSVPVYLTPSYTASAVTWSCS TTTGNEKYVPSECRKVGT >Pil_TUM15069 (SEQ ID NO: 84) MNTMQKGFTLIELMIVVAIIGILAAIAIPAYQDYTIRAKISEGLTLSNGLKTAIAESFQSKGPSSMAC TDATTCASIGASPMDATALAGNKNVASITSSTAGVITIAYKPAVVPNGSNNLTLTPVGADGTTAL DLSAAASAGSQVNWRCGGTGTTVAAKFLPANCRGT >Pil_AI7 (SEQ ID NO: 85) MNAQKGFTLIELMIVVAIIGILAAVAIPAYQDYTTRAKVSEVITAGAACKTSVAEYYQSTGSLPLN TEQAGCSSNATPMVKSLAVASGIITVTASDALAAKFSTSTQNTYVLEPTATTAAAPLTWSCTGSTI EGKYLPAECRGT >Pil_VE-C3 (SEQ ID NO: 86) MKSMQKGFTLIELMIVIAIIGILAAIAIPAYTDYTARAKITEAVGALASAKTSVSEYYTSMGKMPA DAAAAGINTAPAGSYVDNVAYAKTSDTVSTVTATIKNVNSTADTKKFKLTGTGSVAGVTWACA TVDLDQKYLPANCRST >Pil_YH01026 (SEQ ID NO: 87) MNAQKGFTLIELMIVVAIIGILAAIAIPAYQDYTIRAKISEGLTLSNGLKTAIAESFQSKGPSSMECN NAATCALIGASPMDATALGGNKNVTSITSSEAGVITIAYKPAVVPAGANNLLLTPVGADGTTALN LSAAASAGSQVNWQCGGTNGTTVAAKFLPANCRGT Atty. Dkt. No.64100-229640 >Pil_Mo (SEQ ID NO: 57) MNAQKGFTLIELMIVVAIIGILAAIAIPAYQDYTIRAKISEGLTLSNGLKTAIAESFQSKGPSSMACT DATTCASIGASPMDATALAGNKNVASITSDAAGVITIVYKPAVVPTGSNNLTLTPVGADGTTALN LSAAASAGSQVNWRCGGTGTTVAAKFLPANCRGT >Pil_CIP102143 (SEQ ID NO: 88) MIVVAIIGILAAVAIPAYQDYTTRAKVSEVITAGAACKTSVAEYYQSTGSLPVNTEQAGCSSNATP MVKSLAVDKGVITVTASDSLAAKFSTSTKNTYVLRPTATSAAAPLTWSCTGSTIEGKYLPAECRG T >Pil_AI40 (SEQ ID NO: 89) MNAQKGFTLIELMIVVAIIGILAAIAIPAYQDYTTRAKMSEVVNFAAAAKSAVSECAISTGDLDEC DSNQKAGLAPAADLTSTYVESVTVGADGLITLAIQGTNVTALDNGSLTMEPTLDPVAGVTWVCK ISSNTLNKYVPANCRAT >Pil_F78 (SEQ ID NO: 90) MNTVQKGFTLIELMIVIAIIGILAAIAIPAYTDYTVRARVSEAMTTASAMKATVSENIMNAGGTSIV ATNKNCAGIPAFTATKNVATAACTDKTGVILVTTTEAAKSVPLTLTPTYSGGNVSWRCSTTSSFD KYVPSECRST >Pil_S71 (SEQ ID NO: 91) MNAQKGFTLIELMIVVAIIGILAAVAIPAYQDYTTRAKVSEVITAGAACKTSVAEFYQSSGELPGT LEQAGCSSNATPMVASLDVGADGVITVTASTDLAAKFSDSAKNTYVLAPTATTAAAPLTWSCTG STIEGKYLPAECRGT >Pil_ANC4282 (SEQ ID NO: 92) MNAQKGFTLIELMIVVAIIGILAAIAIPAYQDYTTRAKVTEVMNYAAAAKSAVSECLSSTGVTTSC DTNAEAGLEAATSLTSPYVTSVTVGTGGSITAVVKGTNATSGNVALDGASLVLTPALSNAGVAW TCKISNVALNKFVPQTCRST >Pil_72-O-c (SEQ ID NO: 93) MNAQKGFTLIELMIVIAIIGILAAIAIPAYQNYIAKSQAAEAFTLMGGAKTTINSNLQNNSCTNTDD DTQNTVNGKYGVLTIGGAVAQDSNPTAATGCTMSYLFKGTGVSSQLADLVIEAGLNNNGTLAID DTATTVDDELLPKSFAS >Pil_BI730 (SEQ ID NO: 94) MNAQKGFTLIELMIVVAIIGILAAIAIPQYQNYIAKSQVSRAMGETSSVKTAVETCLNEGKGAAGA CPLGITTSNIQATTVAGSAPKADGTNAAELTETTAIVATFGTGAAKTLQETGKNSVTWTRDATGS WKCNSTVQAKYAPAGCSAT >Pil_A3K91 (SEQ ID NO: 95) MNAQKGFTLIELMIVVAIIGILAAIAIPAYQNYIAKSQASEAFTLIDGAKAEVNTNLEGNSCTNATA AKNTIAGKYGSLVIAGTAASDASPTASTGCTLTYTFKGTGVSSQLASKVIGATLLNNGTLTKNAT TTTVDADILPKSFT >Pil_CIP102159 (SEQ ID NO: 96) MNTMQKGFTLIELMIVVAIIGILAAIAIPAYQDYTIRSRVAEALTALSSAKATVSENIANNGGVIAA GACAGYTNQTTATANVASTSCTDTTGVVSATTTTKAGGFVITMTPTVNADKTVVWKCTVPAAS FKYAPAECRGT Atty. Dkt. No.64100-229640 >Pil_junii_65 (SEQ ID NO: 97) MKSMQKGFTLIELMIVVAIIGILAAIAIPAYQDYTTRAKMSEVVNFAAAAKSAVSECAISTGTLSD CNTNAKAGLEAAADLKSTYVESVTVGNNGVITLEIKGTNVTALDAANLTMTPTLDNKAGVSWV CKISSNTLNKYVPANCRST >Pil_YZS-X (SEQ ID NO: 98) MNTVQKGFTLIELMIVVAIIGILAAIAIPAYTDYTARAKVTEAVGALASAKTSVSEYFTSQGVMPT NATQAGINTAPAGQYVSAVAYTKTSDTVSKISATLANINSEANTKTIVLEGTGDTAKGVSWVCK GGTAPGKFLPANCRGT >Pil_T-3-2 (SEQ ID NO: 99) MNAQKGFTLIELMIVIAIIGILAAIAIPAYQNYIAKAQVAEAFTLADGIKTSVGTNLQSGTCFASGA ATAATTDTIEGKYGTATTVADTTTGSNGCGIKYTFKSSGVSNKLTSKVIGMAITENGVLKKSTVT TTDAPADLLPQSFT >Pil_CIP102637 (SEQ ID NO: 100) MECNVLMIVVAIIGILAAIAIPAYTDYTARAKVTEAVGALASAKTSVSEFFTSQGVMPSDADAAGI NKTPAGDYVAGVTYTRTDPTHAVVAVELKNINSDANGTTFQLNATGDTAKGVSWTCSSAGTKP TPEKFLPANCRGT >Pil_Mo_H28 (SEQ ID NO: 58) AYQDYTIRAKISEGLTLSNGLKTAIAESFQSKGPSSMACTDATTCASIGASPMDATALAGNKNVA SITSDAAGVITIVYKPAVVPTGSNNLTLTPVGADGTTALNLSAAASAGSQVNWRCGGTGTTVAA KFLPANCRGT > Pil<sub>20</sub> sequon / Pil_Mo_DSL (SEQ ID NO: 60) CGGTGTTVAAKFLPANCRGT >EPA-Pil_Mo_H28 (SEQ ID NO: 101) MKKIWLALAGLVLAFSASAAEEAFDLWNECAKACVLDLKDGVRSSRMSVDPAIADTNGQGVLH YSMVLEGGNDALKLAIDNALSITSDGLTIRLEGGVEPNKPVRYSYTRQARGSWSLNWLVPIGHEK PSNIKVFIHELNAGNQLSHMSPIYTIEMGDELLAKLARDATFFVRAHESNEMQPTLAISHAGVSVV MAQAQPRREKRWSEWASGKVLCLLDPLDGVYNYLAQQRCNLDDTWEGKIYRVLAGNPAKHDL DIKPTVISHRLHFPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAAR LSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASAD VVSLTCPVAAGECAGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLE ERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIR NGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLTILGWPLA ERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPREDLKAYQDYTIRAKISEGL TLSNGLKTAIAESFQSKGPSSMACTDATTCASIGASPMDATALAGNKNVASITSDAAGVITIVYKP AVVPTGSNNLTLTPVGADGTTALNLSAAASAGSQVNWRCGGTGTTVAAKFLPANCRGT >EPA-Pil20 (SEQ ID NO: 102) MKKIWLALAGLVLAFSASAAEEAFDLWNECAKACVLDLKDGVRSSRMSVDPAIADTNGQGVLH YSMVLEGGNDALKLAIDNALSITSDGLTIRLEGGVEPNKPVRYSYTRQARGSWSLNWLVPIGHEK PSNIKVFIHELNAGNQLSHMSPIYTIEMGDELLAKLARDATFFVRAHESNEMQPTLAISHAGVSVV MAQAQPRREKRWSEWASGKVLCLLDPLDGVYNYLAQQRCNLDDTWEGKIYRVLAGNPAKHDL DIKPTVISHRLHFPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAAR LSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASAD VVSLTCPVAAGECAGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLE ERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIR NGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLTILGWPLA Atty. Dkt. No.64100-229640 ERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPREDLKGGGGCGGTGTTVAA KFLPANCRGT >PglS Sequon (SEQ ID NO: 59) CTGVTQIASGASAATTNVASAQC >EPA_PglS sequon_Pil<sub>20</sub> (SEQ ID NO: 103) MKKIWLALAGLVLAFSASAAEEAFDLWNECAKACVLDLKDGVRSSRMSVDPAIADTNGQGVLH YSMVLEGGNDALKLAIDNALSITSDGLTIRLEGGVEPNKPVRYSYTRQARGSWSLNWLVPIGHEK PSNIKVFIHELNAGNQLSHMSPIYTIEMGDELLAKLARDATFFVRAHESNEMQPTLAISHAGVSVV MAQAQPRREKRWSEWASGKVLCLLDPLDGVYNYLAQQRCNLDDTWEGKIYRVLAGNPAKHDL DIKPTVISHRLHFPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAAR LSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASAD VVSLTCPVAAGECAGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLE ERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDACTGVT QIASGASAATTNVASAQCVRGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLP LRLDAITGPEEEGGRLTILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPG KPPREDLKGGGGCGGTGTTVAAKFLPANCRGT >EPA_PglS sequon 2X (SEQ ID NO: 104) MKKIWLALAGLVLAFSASAAEEAFDLWNECAKACVLDLKDGVRSSRMSVDPAIADTNGQGVLH YSMVLEGGNDALKLAIDNALSITSDGLTIRLEGGVEPNKPVRYSYTRQARGSWSLNWLVPIGHEK PSNIKVFIHELNAGNQLSHMSPIYTIEMGDELLAKLARDATFFVRAHESNEMQPTLAISHAGVSVV MAQAQPRREKRWSEWASGKVLCLLDPLDGVYNYLAQQRCNLDDTWEGKIYRVLAGNPAKHDL DIKPTVISHRLHFPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAAR LSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASAD VVSLTCPVAAGECAGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLE ERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDACTGVT QIASGASAATTNVASAQCVRGRIRNGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLP LRLDAITGPEEECTGVTQIASGASAATTNVASAQCGGRLTILGWPLAERTVVIPSAIPTDPRNVGG DLDPSSIPDKEQAISALPDYASQPGKPPREDLK >Dsba_secretion_signal_sequence (SEQ ID NO: 142) MKKIWLALAGLVLAFSASA 48ZIHG9B5GL_V[V_PUG5 Gg9.., #E9C =8 @A3 *-,$ #9B5 JHYYPLY ^P[OV\[ HU` ZLX\VUZ' FOPZ cIHYLd version has a non-native N-terminal DsbA secretion signal sequence and has a deletion of residue E553 which inactivates the toxin) MKKIWLALAGLVLAFSASAAEEAFDLWNECAKACVLDLKDGVRSSRMSVDPAIADTNGQGVLH YSMVLEGGNDALKLAIDNALSITSDGLTIRLEGGVEPNKPVRYSYTRQARGSWSLNWLVPIGHEK PSNIKVFIHELNAGNQLSHMSPIYTIEMGDELLAKLARDATFFVRAHESNEMQPTLAISHAGVSVV MAQAQPRREKRWSEWASGKVLCLLDPLDGVYNYLAQQRCNLDDTWEGKIYRVLAGNPAKHDL DIKPTVISHRLHFPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAAR LSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAASAD VVSLTCPVAAGECAGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLE ERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIR NGALLRVYVPRSSLPGFYRTGLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLTILGWPLA ERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPREDLK >FlgI_secretion_signal_sequence (SEQ ID NO: 144) MIKFLSALILLLVTTAAQA Atty. Dkt. No.64100-229640 >FlgI_CRM197_G52E (SEQ ID NO: 145) (Inactivated (G52E) form of CRM197 with an N-terminal non- native FlgI secretion signal sequence) MIKFLSALILLLVTTAAQAGADDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNY DDDWKEFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELGLS LTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEINFETRGKRGQ DAMYEYMAQACAGNRVRRSVGSSLSCINLDWDVIRDKTKTKIESLKEHGPIKNKMSESPNKTVS EEKAKQYLEEFHQTALEHPELSELKTVTGTNPVFAGANYAAWAVNVAQVIDSETADNLEKTTA ALSILPGIGSVMGIADGAVHHNTEEIVAQSIALSSLMVAQAIPLVGELVDIGFAAYNFVESIINLFQ VVHNSYNRPAYSPGHKTQPFLHDGYAVSWNTVEDSIIRTGFQGESGHDIKITAENTPLPIAGVLLP TIPGKLDVNKSKTHISVNGRKIRMRCRAIDGDVTFCRPKSPVYVGNGVHANLHVAFHRSSSEKIH SNEISSDSIGVLGYQKTVDHTKVNSKLSLFFEIKS >DsbA_tetanus_toxin_C-fragment_TTc (SEQ ID NO: 146) MKKIWLALAGLVLAFSASAAKNLDCWVDNEEDIDVILKKSTILNLDINNDIISDISGFNSSVITYPD AQLVPGINGKAIHLVNNESSEVIVHKAMDIEYNDMFNNFTVSFWLRVPKVSASHLEQYGTNEYSI ISSMKKHSLSIGSGWSVSLKGNNLIWTLKDSAGEVRQITFRDLPDKFNAYLANKWVFITITNDRLS SANLYINGVLMGSAEITGLGAIREDNNITLKLDRCNNNNQYVSIDKFRIFCKALNPKEIEKLYTSY LSITFLRDFWGNPLRYDTEYYLIPVASSSKDVQLKNITDYMYLTNAPSYTNGKLNIYYRRLYNGL KFIIKRYTPNNEIDSFVKSGDFIKLYVSYNNNEHIVGYPKDGNAFNNLDRILRVGYNAPGIPLYKK MEAVKLRDLKTYSVQLKLYDDKNASLGLVGTHNGQIGNDPNRDILIASNWYFNHLKDKILGCD WYFVPTDEGWTND >DsbA_cholera_toxin_B_subunit (SEQ ID NO: 147) MKKIWLALAGLVLAFSASANGTPQNITDLCAEYHNTQIHTLNDKIFSYTESLAGKREMAIITFKNG ATFQVEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVEKLCVWNNKTPHAIAAISMAN

Atty. Dkt. No.64100-229640

Atty. Dkt. No.64100-229640

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Claims

CLAIMS What is claimed is: 1. A glycoconjugate comprising an oligo- or polysaccharide covalently linked to an acceptor protein, wherein the acceptor protein comprises or consists of a TfpM-associated pilin-like protein or glycosylation fragment thereof and the oligo- or polysaccharide is covalently linked to the TfpM-associated pilin-like protein or glycosylation fragment thereof, and wherein the TfpM-associated pilin-like protein or glycosylation fragment thereof comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine; optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin-like protein C-terminal end; optionally, wherein the acceptor protein is a fusion protein comprising the TfpM- associated pilin-like protein or glycosylation fragment thereof translationally fused to a heterologous carrier protein and the TfpM-associated pilin-like protein or glycosylation fragment thereof is the C-terminus-most sequence of the acceptor protein such that the acceptor protein comprises a C-terminus serine or threonine residue and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine; optionally, wherein the oligo- or polysaccharide comprises a glucose at its reducing end; and/or optionally, wherein the glycoconjugate is immunogenic. 2. The glycoconjugate of Claim 1, wherein the TfpM-associated pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. Atty. Dkt. No.64100-229640 3. The glycoconjugate of Claim 1, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) PilMo (SEQ ID NO: 57), (ii) PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3.1$% VY #PPP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ .)"% /)"% 0)"% 0."% 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein comprises a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Pil<sub>AI40</sub> (SEQ ID NO: 89), Pil<sub>F78</sub> (SEQ ID NO: 90), Pil<sub>S71</sub> (SEQ ID NO: 91), Pil<sub>ANC4282</sub> (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), Pil<sub>CIP102159</sub> (SEQ ID NO: 96), Pil<sub>junii-65</sub> (SEQ ID NO: 97), Pil<sub>YZS-X</sub> (SEQ ID NO: 98), Pil<sub>T-3-2</sub> (SEQ ID NO: 99), and Pil<sub>CIP102637</sub> (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of Pil<sub>DSM16617</sub> (SEQ ID NO: 82), Pil<sub>ZZC3-9</sub> (SEQ ID NO: 83), Pil<sub>TUM15069</sub> (SEQ ID NO: 84), Pil<sub>AI7</sub> (SEQ ID NO: 85), Pil<sub>VE-C3</sub> (SEQ ID NO: 86), Pil<sub>YH01026</sub> (SEQ ID NO: 87), Pil<sub>CIP102143</sub> (SEQ ID NO: 88), Pil<sub>AI40</sub> (SEQ ID NO: 89), Pil<sub>F78</sub> (SEQ ID NO: 90), Pil<sub>S71</sub> (SEQ ID NO: 91), Pil<sub>ANC4282</sub> (SEQ ID NO: 92), Pil<sub>72-O-c</sub> (SEQ ID NO: 93), Pil<sub>BI730</sub> (SEQ ID NO: 94), Pil<sub>A3K91</sub> (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), Pil<sub>CIP102637</sub> (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), Atty. Dkt. No.64100-229640 optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil19 (SEQ ID NO: 133), Pil18 (SEQ ID NO: 134), Pil17 (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil8 (SEQ ID NO: 141), Pil7 (SEQ ID NO: 113), Pil6 (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil<sub>14[A]</sub> (SEQ ID NO: 172), Pil<sub>13[A]</sub> (SEQ ID NO: 173), Pil<sub>12[A]</sub> (SEQ ID NO: 174), Pil<sub>11[A]</sub> (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil<sub>7[A]</sub> (SEQ ID NO: 179), Pil<sub>6[A]</sub> (SEQ ID NO: 180), Pil<sub>5[A]</sub> (SEQ ID NO: 181), or Pil<sub>4[A]</sub> (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil<sub>20S</sub> (SEQ ID NO: 148), Pil<sub>19S</sub> (SEQ ID NO: 149), Pil<sub>18S</sub> (SEQ ID NO: 150), Pil<sub>17S</sub> (SEQ ID NO: 151), Pil<sub>16S</sub> (SEQ ID NO: 152), Pil<sub>15S</sub> (SEQ ID NO: 153), Pil<sub>14S</sub> (SEQ ID NO: 154), Pil<sub>13S</sub> (SEQ ID NO: 155), Pil<sub>12S</sub> (SEQ ID NO: 156), Pil<sub>11S</sub> (SEQ ID NO: 157), Pil<sub>10S</sub> (SEQ ID NO: 158), Pil<sub>9S</sub> (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil<sub>5S</sub> (SEQ ID NO: 163), Pil<sub>4S</sub> (SEQ ID NO: 164), or Pil<sub>3S</sub> (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or Atty. Dkt. No.64100-229640 five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 4. The glycoconjugate of Claim 1, wherein the acceptor protein is a fusion protein comprising a heterologous carrier protein and the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. 5. The glycoconjugate of Claim 1, wherein the acceptor protein is a fusion protein and the fusion protein further comprises an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus, optionally wherein the additional glycosylation sequence is an internal glycosylation fragment of ComP, further optionally, wherein the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13; optionally, wherein the additional glycosylation sequence is also covalently linked to an oligo- or polysaccharide. 6. The glycoconjugate of Claim 5, wherein the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more additional glycosylation sequences; optionally, wherein the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five additional glycosylation sequences; optionally, wherein the additional glycosylation sequences are identical; optionally, wherein the additional glycosylation sequences differ from each other; and/or optionally, wherein at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other; and/or optionally, wherein none of the additional glycosylation sequences are the same. Atty. Dkt. No.64100-229640 7. The glycoconjugate of Claim 1, wherein the oligo- or polysaccharide covalently linked to the pilin-like protein or glycosylation fragment thereof has a size of at least three repeating units of oligo- or polysaccharide structure and/or has a size of at least ten monosaccharides. 8. The glycoconjugate of Claim 1, (i) wherein the oligo- or polysaccharide is produced by bacteria of the genus Streptococcus and the polysaccharide is capsular polysaccharide, optionally, wherein is S. pneumoniae or S. agalactiae, and optionally, wherein the S. agalactiae capsular polysaccharide is Ia, Ib, II, III, IV, V, VI, VII, VIII, or IX; (ii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Klebsiella and the polysaccharide is a capsular polysaccharide or O-antigen polysaccharide, optionally wherein the bacteria is K. pneumoniae; or (iii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Salmonella and the polysaccharide is O-antigen polysaccharide; optionally wherein the bacteria is S. enterica and the S. enterica polysaccharide is a Group B O-antigen. 9. The glycoconjugate of Claim 1, wherein the glycoconjugate is produced in vivo; optionally, in a bacterial cell; optionally, in Escherichia coli; optionally, in a bacterium from the genus Klebsiella; and/or optionally, wherein the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 10. The glycoconjugate of Claim 1, wherein the bioconjugate is a conjugate vaccine that induces an immune response when administered to a subject; optionally, wherein the immune response elicits long term memory (memory B and T cells), is an antibody response, and is optionally a serotype-specific antibody response; optionally, wherein the antibody response is an IgG or IgM response; optionally, wherein the antibody response is an IgG response; optionally an IgG1 response; and/or Atty. Dkt. No.64100-229640 optionally, wherein the conjugate vaccine generates immunological memory in a subject administered the vaccine. 11. A pilin-like-protein glycosylation fragment comprising or consisting of an isolated fragment of a TfpM-associated pilin-like protein, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) PilMo (SEQ ID NO: 57), (ii) PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3.1$% VY #PPP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine, (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), Pil_VE-C3 (SEQ ID NO: 86), Pil_YH01026 (SEQ ID NO: 87), Pil_CIP102143 (SEQ ID NO: 88), Pil_AI40 (SEQ ID NO: 89), Pil_F78 (SEQ ID NO: 90), Pil_S71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil_72-O-c (SEQ ID NO: 93), Pil_BI730 (SEQ ID NO: 94), Pil_A3K91 (SEQ ID NO: 95), Pil_CIP102159 (SEQ ID NO: 96), Pil_junii-65 (SEQ ID NO: 97), Pil_YZS-X (SEQ ID NO: 98), Pil_T-3-2 (SEQ ID NO: 99), and Pil_CIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), Pil_TUM15069 (SEQ ID NO: 84), Pil_AI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Pil_AI40 (SEQ ID NO: 89), Pil_F78 (SEQ ID NO: 90), Pil_S71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- Atty. Dkt. No.64100-229640 terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil19 (SEQ ID NO: 133), Pil18 (SEQ ID NO: 134), Pil17 (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil8 (SEQ ID NO: 141), Pil7 (SEQ ID NO: 113), Pil6 (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil_20[A] (SEQ ID NO: 166), Pil_19[A] (SEQ ID NO: 167), Pil_18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil_14[A] (SEQ ID NO: 172), Pil_13[A] (SEQ ID NO: 173), Pil_12[A] (SEQ ID NO: 174), Pil_11[A] (SEQ ID NO: 175), Pil_10[A] (SEQ ID NO: 176), Pil_9[A] (SEQ ID NO: 177), Pil_8[A] (SEQ ID NO: 178), Pil_7[A] (SEQ ID NO: 179), Pil_6[A] (SEQ ID NO: 180), Pil_5[A] (SEQ ID NO: 181), or Pil_4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil_20S (SEQ ID NO: 148), Pil_19S (SEQ ID NO: 149), Pil_18S (SEQ ID NO: 150), Pil_17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ ID NO: 155), Pil_12S (SEQ ID NO: 156), Pil_11S (SEQ ID NO: 157), Pil_10S (SEQ ID NO: 158), Pil_9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Atty. Dkt. No.64100-229640 Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 12. The pilin-like-protein glycosylation fragment of Claim 11, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 13. A fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof translationally fused to a heterologous carrier protein, wherein the TfpM-associated pilin-like protein or glycosylation fragment comprises a C- terminus serine or threonine residue, wherein the TfpM-associated pilin-like protein or glycosylation fragment is the C- terminus-most sequence of the fusion protein, and wherein the fusion protein comprises a C-terminus serine or threonine residue; optionally, wherein the fusion protein is glycosylated by an olio- or polysaccharide covalently linked to the C-terminus serine or threonine; optionally, wherein the fusion protein is glycosylated by an oligo- or polysaccharide comprising glucose at its reducing end covalently linked to the C-terminus serine or threonine. 14. The fusion protein of Claim 13, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 Atty. Dkt. No.64100-229640 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 15. The fusion protein of Claim 13, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) PilMo (SEQ ID NO: 57), (ii) PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3.1$% VY #PP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of Pil<sub>DSM16617</sub> (SEQ ID NO: 82), Pil<sub>ZZC3-9</sub> (SEQ ID NO: 83), Pil<sub>TUM15069</sub> (SEQ ID NO: 84), Pil<sub>AI7</sub> (SEQ ID NO: 85), Pil<sub>VE-C3</sub> (SEQ ID NO: 86), Pil<sub>YH01026</sub> (SEQ ID NO: 87), Pil<sub>CIP102143</sub> (SEQ ID NO: 88), Pil<sub>AI40</sub> (SEQ ID NO: 89), Pil<sub>F78</sub> (SEQ ID NO: 90), Pil<sub>S71</sub> (SEQ ID NO: 91), Pil<sub>ANC4282</sub> (SEQ ID NO: 92), Pil<sub>72-O-c</sub> (SEQ ID NO: 93), Pil<sub>BI730</sub> (SEQ ID NO: 94), Pil<sub>A3K91</sub> (SEQ ID NO: 95), Pil<sub>CIP102159</sub> (SEQ ID NO: 96), Pil<sub>junii-65</sub> (SEQ ID NO: 97), Pil<sub>YZS-X</sub> (SEQ ID NO: 98), Pil<sub>T-3-2</sub> (SEQ ID NO: 99), and Pil<sub>CIP102637</sub> (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), Pil<sub>VE-C3</sub> (SEQ ID NO: 86), Pil<sub>YH01026</sub> (SEQ ID NO: 87), Pil<sub>CIP102143</sub> (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or Atty. Dkt. No.64100-229640 truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil19 (SEQ ID NO: 133), Pil18 (SEQ ID NO: 134), Pil17 (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil8 (SEQ ID NO: 141), Pil7 (SEQ ID NO: 113), Pil6 (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil<sub>17[A]</sub> (SEQ ID NO: 169), Pil<sub>16[A]</sub> (SEQ ID NO: 170), Pil<sub>15[A]</sub> (SEQ ID NO: 171), Pil<sub>14[A]</sub> (SEQ ID NO: 172), Pil<sub>13[A]</sub> (SEQ ID NO: 173), Pil<sub>12[A]</sub> (SEQ ID NO: 174), Pil<sub>11[A]</sub> (SEQ ID NO: 175), Pil<sub>10[A]</sub> (SEQ ID NO: 176), Pil<sub>9[A]</sub> (SEQ ID NO: 177), Pil<sub>8[A]</sub> (SEQ ID NO: 178), Pil<sub>7[A]</sub> (SEQ ID NO: 179), Pil<sub>6[A]</sub> (SEQ ID NO: 180), Pil<sub>5[A]</sub> (SEQ ID NO: 181), or Pil<sub>4[A]</sub> (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil<sub>20S</sub> (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil<sub>16S</sub> (SEQ ID NO: 152), Pil<sub>15S</sub> (SEQ ID NO: 153), Pil<sub>14S</sub> (SEQ ID NO: 154), Pil<sub>13S</sub> (SEQ ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID Atty. Dkt. No.64100-229640 NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 16. The fusion protein of Claim 13, wherein the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. 17. The fusion protein of Claim 13, wherein the fusion protein further comprises an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus, optionally wherein the additional glycosylation sequence is an internal glycosylation fragment of ComP, further optionally, wherein the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13; optionally, wherein the additional glycosylation sequence is also covalently linked to an oligo- or polysaccharide. 18. The fusion protein of Claim 17, wherein the fusion protein comprises two or more, three or more, four or more, five or more, six or more, eight or more, ten or more, fifteen or more, or twenty or more additional glycosylation sequence; optionally, wherein the fusion protein does not comprise more than two, more than three, more than five, more than ten, more than fifteen, more than twenty, or more than twenty five additional glycosylation sequence; optionally, wherein the additional glycosylation sequence are identical; optionally, wherein the additional glycosylation sequence differ from each other; and/or optionally, wherein at least three, at least four, or at least five of the additional glycosylation sequences all differ from each other; and/or optionally, wherein none of the additional glycosylation sequence are the same. Atty. Dkt. No.64100-229640 19. The fusion protein of Claim 13, wherein the oligo- or polysaccharide covalently linked to the pilin-like protein or glycosylation fragment thereof has a size of at least three repeating units of oligo- or polysaccharide structure and/or has a size of at least ten monosaccharides. 20. The fusion protein of Claim 13, (i) wherein the oligo- or polysaccharide is produced by bacteria of the genus Streptococcus and the polysaccharide is capsular polysaccharide, optionally, wherein is S. pneumoniae or S. agalactiae, and optionally, wherein the S. agalactiae capsular polysaccharide is Ia, Ib, II, III, IV, V, VI, VII, VIII, or IX; (ii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Klebsiella and the polysaccharide is a capsular polysaccharide or O-antigen polysaccharide, optionally wherein the bacteria is K. pneumoniae; or (iii) wherein the oligo- or polysaccharide is produced by bacteria of the genus Salmonella and the polysaccharide is O-antigen polysaccharide; optionally wherein the bacteria is S. enterica and the S. enterica polysaccharide is a Group B O-antigen. 21. The fusion protein of Claim 13, wherein the glycosylated fusion protein is produced in vivo; optionally, in a bacterial cell; optionally, in Escherichia coli; optionally, in a bacterium from the genus Klebsiella; and/or optionally, wherein the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 22. The fusion protein of Claim 13, wherein when the fusion protein is a vaccine that induces an immune response when administered to a subject; optionally, wherein the immune response elicits long term memory (memory B and T cells), is an antibody response, and is optionally a serotype-specific antibody response; optionally, wherein the antibody response is an IgG or IgM response; Atty. Dkt. No.64100-229640 optionally, wherein the antibody response is an IgG response; optionally an IgG1 response; and/or optionally, wherein the fusion protein generates immunological memory in a subject administered the vaccine. 23. A method of producing a glycoconjugate, the method comprising covalently linking an oligo- or polysaccharide to an acceptor protein comprising or consisting of a TfpM-associated pilin-like protein or glycosylation fragment thereof using a TfpM oligosaccharyltransferase (OTase); wherein the pilin-like protein or glycosylation fragment comprises a C-terminus serine or threonine residue, the acceptor protein comprises a C-terminus serine or threonine residue, and the oligo- or polysaccharide is covalently linked to the C-terminus serine or threonine residue of the acceptor protein; optionally, wherein the oligo- or polysaccharide comprises a glucose at its reducing end; optionally, wherein the acceptor protein is a fusion protein of any one of Claims 13 to 22; optionally, wherein the method is a method of in vivo conjugation of an oligo- or polysaccharide to an acceptor protein; and/or optionally, wherein the glycoconjugate is immunogenic. 24. The method of Claim 23, wherein the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpM<sub>Mo</sub> (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpM<sub>AI7</sub> (SEQ ID NO: 66), TfpM<sub>VE-C3</sub> (SEQ ID NO: 67), TfpM<sub>YH01026</sub> (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 Atty. Dkt. No.64100-229640 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), or TfpM72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpMMo (SEQ ID NO: 56). 25. The method of Claim 23, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 26. The method of Claim 23, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) Pil<sub>Mo</sub> (SEQ ID NO: 57), (ii) Pil<sub>Mo</sub> lacking amino acids corresponding to residues 1–28 (Pil<sub>Mo</sub>g+1% E9C =8 @A3.1$% VY #PP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), Pil_TUM15069 (SEQ ID NO: 84), Pil_AI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), and PilCIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of PilDSM16617 (SEQ ID NO: 82), PilZZC3-9 (SEQ ID NO: 83), PilTUM15069 (SEQ ID NO: 84), PilAI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID Atty. Dkt. No.64100-229640 NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil72-O-c (SEQ ID NO: 93), PilBI730 (SEQ ID NO: 94), PilA3K91 (SEQ ID NO: 95), PilCIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C- terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C- terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil₁₉ (SEQ ID NO: 133), Pil₁₈ (SEQ ID NO: 134), Pil₁₇ (SEQ ID NO: 135), Pil₁₆ (SEQ ID NO: 136), Pil₁₅ (SEQ ID NO: 109), Pil₁₄ (SEQ ID NO: 137), Pil₁₃ (SEQ ID NO: 110), Pil₁₂ (SEQ ID NO: 138), Pil₁₁ (SEQ ID NO: 139), Pil₁₀ (SEQ ID NO: 112), Pil₉ (SEQ ID NO: 140), Pil₈ (SEQ ID NO: 141), Pil₇ (SEQ ID NO: 113), Pil₆ (SEQ ID NO: 114), Pil₅ (SEQ ID NO: 115), Pil₄ (SEQ ID NO: 116), or Pil₃ (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil_17[A] (SEQ ID NO: 169), Pil_16[A] (SEQ ID NO: 170), Pil_15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil12[A] (SEQ ID NO: 174), Pil11[A] (SEQ ID NO: 175), Pil10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil5[A] (SEQ ID NO: 181), or Pil4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil20S (SEQ ID NO: 148), Pil19S (SEQ ID NO: 149), Pil18S (SEQ ID NO: 150), Pil17S (SEQ ID NO: 151), Pil16S (SEQ ID NO: 152), Pil15S (SEQ ID NO: 153), Pil14S (SEQ ID NO: 154), Pil13S (SEQ Atty. Dkt. No.64100-229640 ID NO: 155), Pil12S (SEQ ID NO: 156), Pil11S (SEQ ID NO: 157), Pil10S (SEQ ID NO: 158), Pil9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil4S (SEQ ID NO: 164), or Pil3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C- terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil_4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 27. The method of Claim 23, wherein the acceptor protein is a fusion protein comprising a heterologous carrier protein and the carrier protein is selected from the group consisting of Pseudomonas aeruginosa Exotoxin A (EPA), CRM197, cholera toxin B subunit, tetanus toxin C fragment, and a fragment of any thereof. 28. The method of Claim 23, wherein the acceptor protein is a fusion protein comprising an additional glycosylation sequence of an OTase other than TfpM oligosaccharyltransferase (OTase) in addition to the TfpM-associated pilin-like-protein glycosylation fragment located at its C-terminus, and the method further comprises covalently linking an oligo- or polysaccharide to the additional glycosylation sequence with an OTase other than TfpM OTase; optionally, wherein the acceptor protein is a fusion protein comprising a ComP glycosylation fragment, and the method further comprises covalently linking an oligo- or polysaccharide to the ComP glycosylation fragment using a PglS OTase, optionally, wherein the ComP glycosylation fragment is an internal glycosylation fragment of ComP, further optionally, wherein the ComP glycosylation fragment comprises or consists of CTGVTQIASGASAATTNVASAQC (SEQ ID NO: 59) or a fragment thereof comprising at least the amino acids ASA in positions 11-13. Atty. Dkt. No.64100-229640 29. The method of Claim 23, wherein the conjugation occurs in a host cell. 30. The method of Claim 29, wherein the host cell is a bacterial cell; optionally, in Escherichia coli; optionally, in a bacterium from the genus Klebsiella; or optionally, wherein the bacterial species is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 31. The method of Claim 29, comprising culturing a host cell that comprises: (a) a genetic cluster encoding for the proteins required to synthesize the oligo- or polysaccharide; (b) a TfpM OTase; and (3) the acceptor protein. 32. The method of Claim 23, wherein the method produces a conjugate vaccine. 33. A host cell comprising (a) a genetic cluster encoding for the proteins required to synthesize an oligo- or polysaccharide; (b) a TfpM OTase; and (3) an acceptor protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof. 34. The host cell of Claim 33, wherein the acceptor protein is a fusion protein. 35. The host cell of Claim 33, wherein the host cell comprises a nucleic acid encoding the TfpM OTase; and/or wherein the host cell comprises a nucleic acid encoding the acceptor protein; optionally, wherein the TfpM OTase and the acceptor protein are encoded by the same nucleic acid. 36. An isolated nucleic acid encoding the pilin-like-protein glycosylation fragment of Claim 11 or 12 and/or the fusion protein of any one of Claims 13 to 22. 37. The isolated nucleic acid of Claim 36, wherein the nucleic acid is a vector. Atty. Dkt. No.64100-229640 38. A host cell comprising the isolated nucleic acid of Claim 36, optionally, wherein the host cell is a bacterial cell; and further: optionally, wherein the host cell is Escherichia coli; optionally, wherein the host cell is from the genus Klebsiella; or optionally, wherein the host cell is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 39. A composition comprising the conjugate vaccine of Claim 10 or the fusion protein of Claim 22, and an adjuvant and/or carrier. 40. A method of inducing a host immune response against a bacterial pathogen, the method comprising administering to a subject in need of the immune response an effective amount of the conjugate vaccine of Claim 10, the fusion protein of Claim 22, or the composition of Claim 39. 41. The method of Claim 40, wherein the immune response is an antibody response; wherein the immune response is selected from the group consisting of an innate response, an adaptive response, a humoral response, an antibody response, cell mediated response, a B cell response, a T cell response, cytokine upregulation or downregulation, immune system cross-talk, and a combination of two or more of said immune responses; and/or wherein the immune response is selected from the group consisting of an innate response, a humoral response, an antibody response, a T cell response, and a combination of two or more of said immune responses. 42. A method of preventing or treating a bacterial disease and/or infection in a subject comprising administering to a subject in need thereof the conjugate vaccine of Claim 10, the fusion protein of Claim 22, or the composition of Claim 39; optionally, wherein the subject is a human. 43. The method of Claim 42, wherein the infection is a localized or systemic infection of skin, soft tissue, blood, or an organ, or is auto-immune in nature; wherein the disease is pneumonia; and/or Atty. Dkt. No.64100-229640 wherein the infection is a systemic infection and/or an infection of the blood. 44. The method of any one of claims 40 to 43, wherein the conjugate vaccine, the fusion protein, or the composition is administered via intramuscular injection, intradermal injection, intraperitoneal injection, subcutaneous injection, intravenous injection, oral administration, mucosal administration, intranasal administration, or pulmonary administration. 45. A method of producing a pneumococcal conjugate vaccine against pneumococcal infection, the method comprising: (a) isolating the glycoconjugate of any one of Claims 1 to 10 or a glycosylated fusion protein of any one of Claims 13 to 22; and (b) combining the isolated glycoconjugate or isolated glycosylated fusion protein with an adjuvant and/or carrier. 46. The glycoconjugate, glycosylated fusion protein, or conjugate vaccine of any of the above claims for use in inducing a host immune response against a bacterial pathogen and/or preventing or treating a bacterial disease and/or infection in a subject. 47. A recombinant nucleic acid construct comprising a nucleotide sequence encoding a TfpM oligosaccharyltransferase (OTase) operably linked to at least one heterologous transcriptional regulatory sequence. 48. The recombinant construct of Claim 47, wherein the TfpM OTase comprises at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81); Atty. Dkt. No.64100-229640 optionally, wherein the TfpM OTase is TfpMMo (SEQ ID NO: 56), TfpMDSM16617 (SEQ ID NO: 63), TfpMZZC3 (SEQ ID NO: 64), TfpMTUM15069 (SEQ ID NO: 65), TfpMAI7 (SEQ ID NO: 66), TfpMVE-C3 (SEQ ID NO: 67), TfpMYH01026 (SEQ ID NO: 68), TfpMCIP102143 (SEQ ID NO: 69), TfpMAI40 (SEQ ID NO: 70), TfpMF78 (SEQ ID NO: 71), TfpMS71 (SEQ ID NO: 72), TfpMANC4282 (SEQ ID NO: 73), TfpMCIP102159 (SEQ ID NO: 74), TfpMjunii-65 (SEQ ID NO: 75), TfpMYZS-X (SEQ ID NO: 76), TfpMCIP102637 (SEQ ID NO: 77), TfpMT-3-2 (SEQ ID NO: 78), TfpMBI730 (SEQ ID NO: 79), TfpMA3K91 (SEQ ID NO: 80), and/or TfpM72-O-c (SEQ ID NO: 81); optionally, wherein the TfpM OTase is TfpMMo (SEQ ID NO: 56). 49. The recombinant construct of Claim 47 or 48, wherein the heterologous transcriptional regulatory sequence is a promotor sequence. 50. The recombinant construct of any one of Claims 47 to 49, further comprising a nucleotide sequence encoding a TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof operably linked to the nucleotide sequence encoding the a TfpM OTase; optionally, wherein the coding sequence of the TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof is within 2, 5, 10, 20, 30, 40, or 50 nucleotides of the sequence encoding the TfpM OTase; optionally, wherein the coding sequence of the TfpM-associated pilin-like protein or glycosylation fragment thereof or a fusion protein comprising a TfpM-associated pilin-like protein or glycosylation fragment thereof overlaps the operably linked nucleotide sequence encoding the a TfpM OTase. 51. The recombinant construct of Claim 50, wherein the pilin-like-protein glycosylation fragment has a length of from 3 to 139 amino acids in length, has a length of from 20 to 139 amino acids in length, has a length of from 116 to 139 amino acids in length, has a length of from 3 to 22 amino acids in length, has a length of from 10 to 22 amino acids in length, has a length of from 11 to 22 amino acids in length, has a length of from 5 to 21 amino acids in length, has a length of from 10 to 21 amino acids in length, or has a length of from 11 to 21 amino acids in length, Atty. Dkt. No.64100-229640 wherein the TfpM-associated pilin-like protein glycosylation fragment comprises a C- terminus serine or threonine residue. 52. The recombinant construct of Claim 50 or 51, (a) wherein the TfpM-associated pilin-like-protein or glycosylation fragment thereof is: (i) PilMo (SEQ ID NO: 57), (ii) PilMo lacking amino acids corresponding to residues 1–28 (PilMog+1% E9C =8 @A3.1$% VY #PP$ H WVS`WLW[PKL JVTWYPZPUN H[ SLHZ[ JVTWYPZLZ H[ SLHZ[ .)"% 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 57 or SEQ ID NO: 58, wherein the TfpM-associated pilin-like protein contains a C-terminus serine or threonine residue, optionally, wherein the C-terminus threonine is substituted with serine; (b) wherein the TfpM-associated pilin-like-protein is selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), Pil_TUM15069 (SEQ ID NO: 84), Pil_AI7 (SEQ ID NO: 85), PilVE-C3 (SEQ ID NO: 86), PilYH01026 (SEQ ID NO: 87), PilCIP102143 (SEQ ID NO: 88), Pil_AI40 (SEQ ID NO: 89), Pil_F78 (SEQ ID NO: 90), Pil_S71 (SEQ ID NO: 91), Pil_ANC4282 (SEQ ID NO: 92), Pil_72-O-c (SEQ ID NO: 93), Pil_BI730 (SEQ ID NO: 94), Pil_A3K91 (SEQ ID NO: 95), Pil_CIP102159 (SEQ ID NO: 96), Pil_junii-65 (SEQ ID NO: 97), Pil_YZS-X (SEQ ID NO: 98), Pil_T-3-2 (SEQ ID NO: 99), and Pil_CIP102637 (SEQ ID NO: 100); (c) wherein the TfpM-associated pilin-like-protein or the pilin-like-protein glycosylation fragment comprises or consists of an amino acid sequence selected from the group consisting of Pil_DSM16617 (SEQ ID NO: 82), Pil_ZZC3-9 (SEQ ID NO: 83), Pil_TUM15069 (SEQ ID NO: 84), Pil_AI7 (SEQ ID NO: 85), Pil_VE-C3 (SEQ ID NO: 86), Pil_YH01026 (SEQ ID NO: 87), Pil_CIP102143 (SEQ ID NO: 88), PilAI40 (SEQ ID NO: 89), PilF78 (SEQ ID NO: 90), PilS71 (SEQ ID NO: 91), PilANC4282 (SEQ ID NO: 92), Pil_72-O-c (SEQ ID NO: 93), Pil_BI730 (SEQ ID NO: 94), Pil_A3K91 (SEQ ID NO: 95), Pil_CIP102159 (SEQ ID NO: 96), Piljunii-65 (SEQ ID NO: 97), PilYZS-X (SEQ ID NO: 98), PilT-3-2 (SEQ ID NO: 99), PilCIP102637 (SEQ ID NO: 100), and a fragment of any thereof that contains a C-terminus serine or threonine residue, or a variant wherein the C-terminus threonine is substituted with serine, optionally, wherein the TfpM-associated pilin-like protein glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end; and/or (d) wherein the pilin-like-protein glycosylation fragment comprises or consists of the PilMo pilin disulfide loop region (PilMo_DSL, also referred to as Pil20; SEQ ID NO: 60) or truncated derivatives thereof comprising at least the last three amino acids from the pilin C-terminal end or a variant wherein the C-terminus threonine is substituted with serine (SEQ ID NO: 148), Atty. Dkt. No.64100-229640 optionally, (e) wherein the pilin-like-protein glycosylation fragment consists of Pil20 (SEQ ID NO: 60), Pil19 (SEQ ID NO: 133), Pil18 (SEQ ID NO: 134), Pil17 (SEQ ID NO: 135), Pil16 (SEQ ID NO: 136), Pil15 (SEQ ID NO: 109), Pil14 (SEQ ID NO: 137), Pil13 (SEQ ID NO: 110), Pil12 (SEQ ID NO: 138), Pil11 (SEQ ID NO: 139), Pil10 (SEQ ID NO: 112), Pil9 (SEQ ID NO: 140), Pil8 (SEQ ID NO: 141), Pil7 (SEQ ID NO: 113), Pil6 (SEQ ID NO: 114), Pil5 (SEQ ID NO: 115), Pil4 (SEQ ID NO: 116), or Pil3 (SEQ ID NO: 117), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20[A] (SEQ ID NO: 166), Pil19[A] (SEQ ID NO: 167), Pil18[A] (SEQ ID NO: 168), Pil17[A] (SEQ ID NO: 169), Pil16[A] (SEQ ID NO: 170), Pil15[A] (SEQ ID NO: 171), Pil14[A] (SEQ ID NO: 172), Pil13[A] (SEQ ID NO: 173), Pil_12[A] (SEQ ID NO: 174), Pil_11[A] (SEQ ID NO: 175), Pil_10[A] (SEQ ID NO: 176), Pil9[A] (SEQ ID NO: 177), Pil8[A] (SEQ ID NO: 178), Pil7[A] (SEQ ID NO: 179), Pil6[A] (SEQ ID NO: 180), Pil_5[A] (SEQ ID NO: 181), or Pil_4[A] (SEQ ID NO: 182), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus threonine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C- terminal end; or optionally, (f) wherein the pilin-like-protein glycosylation fragment consists of Pil_20S (SEQ ID NO: 148), Pil_19S (SEQ ID NO: 149), Pil_18S (SEQ ID NO: 150), Pil_17S (SEQ ID NO: 151), Pil_16S (SEQ ID NO: 152), Pil_15S (SEQ ID NO: 153), Pil_14S (SEQ ID NO: 154), Pil_13S (SEQ ID NO: 155), Pil_12S (SEQ ID NO: 156), Pil_11S (SEQ ID NO: 157), Pil_10S (SEQ ID NO: 158), Pil_9S (SEQ ID NO: 159), Pil8S (SEQ ID NO: 160), Pil7S (SEQ ID NO: 161), Pil6S (SEQ ID NO: 162), Pil5S (SEQ ID NO: 163), Pil_4S (SEQ ID NO: 164), or Pil_3S (SEQ ID NO: 165), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining the C-terminus serine, further optionally, wherein the glycosylation fragment comprises at least the last three amino acids from the pilin C-terminal end, further optionally, wherein the pilin-like-protein glycosylation fragment consists of Pil20S[A] (SEQ ID NO: 183), Pil19S[A] (SEQ ID NO: 184), Pil18S[A] (SEQ ID NO: 185), Pil17S[A] (SEQ ID NO: 186), Pil16S[A] (SEQ ID NO: 187), Pil15S[A] (SEQ ID NO: 188), Pil14S[A] (SEQ ID NO: 189), Pil13S[A] (SEQ ID NO: 190), Pil12S[A] (SEQ ID NO: 191), Pil11S[A] (SEQ ID NO: 192), Pil10S[A] (SEQ ID NO: 193), Pil9S[A] (SEQ ID NO: 194), Pil8S[A] (SEQ ID NO: 195), Pil7S[A] (SEQ ID NO: 196), Pil6S[A] (SEQ ID NO: 197), Pil5S[A] (SEQ ID NO: 198), or Pil4S[A] (SEQ ID NO: 199), or a variant thereof having one, two, three, four, or five amino acid substitutions and maintaining Atty. Dkt. No.64100-229640 the C-terminus serine, wherein the glycosylation fragment comprises at least the last four amino acids from the pilin C-terminal end. 53. The recombinant construct of any one of Claims 50 to 52, wherein the fusion protein is a fusion protein of any one of Claims 13 to 22. 54. The recombinant construct of any one of Claims 50 to 53, further comprising a nucleotide sequence encoding a PglS OTase operably linked to the TpfM OTase; optionally, wherein the coding sequence of the PglS OTase is within 10, 20, 30, 40, 50, 75, or 100 nucleotides of the sequence encoding the TfpM OTase. 55. A vector comprising the recombinant nucleic acid construct of any one of Claims 47 to 54. 56. A host cell comprising the recombinant nucleic acid construct of any one of Claims 47 to 54, or the vector of Claim 55; optionally, wherein the host cell is a bacterial cell; and further: optionally, wherein the host cell is Escherichia coli; optionally, wherein the host cell is from the genus Klebsiella; or optionally, wherein the host cell is K. pneumoniae, K. varricola, K. michinganenis, or K. oxytoca. 57. A method for producing a TfpM OTase, the method comprising culturing the host cell of Claim 56, wherein said vector of Claim 54 is an expression vector, and recovering the TfpM OTase. 58. A glycoconjugate comprising an oligo- or polysaccharide covalently linked to an acceptor protein, wherein the acceptor protein comprises means for being covalently linked to the oligo- or polysaccharide by a TfpM OTase. 59. The glycoconjugate of claim 58, wherein the glycoconjugate is the glycoconjugate of any one of Claims 1 to 10. Atty. Dkt. No.64100-229640 60. A fusion protein comprising means for being covalently linked to an oligo- or polysaccharide by a TfpM OTase, wherein said means is translationally fused to a heterologous carrier protein, wherein the means is located at the C-terminus-most sequence of the fusion protein. 61. The fusion protein of Claim 60, wherein the fusion protein is the fusion protein of any one of Claims 13 to 22. 62. The glycoconjugate of Claim 58 or 59 or the fusion protein of Claim 60 or 61, for use in any of the methods above. Atty. Dkt. No.64100-229640