WO2020067546A1 - Fil composite, son procédé de production et tissu - Google Patents

Fil composite, son procédé de production et tissu Download PDF

Info

Publication number
WO2020067546A1
WO2020067546A1 PCT/JP2019/038427 JP2019038427W WO2020067546A1 WO 2020067546 A1 WO2020067546 A1 WO 2020067546A1 JP 2019038427 W JP2019038427 W JP 2019038427W WO 2020067546 A1 WO2020067546 A1 WO 2020067546A1
Authority
WO
WIPO (PCT)
Prior art keywords
amino acid
seq
fibroin
fiber
artificial
Prior art date
Application number
PCT/JP2019/038427
Other languages
English (en)
Japanese (ja)
Inventor
明彦 尾関
佑之介 安部
Original Assignee
Ｓｐｉｂｅｒ株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ｓｐｉｂｅｒ株式会社 filed Critical Ｓｐｉｂｅｒ株式会社
Publication of WO2020067546A1 publication Critical patent/WO2020067546A1/fr

Links

Images

Classifications

    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F4/00—Monocomponent artificial filaments or the like of proteins; Manufacture thereof
    • D01F4/02—Monocomponent artificial filaments or the like of proteins; Manufacture thereof from fibroin
    • D—TEXTILES; PAPER
    • D02—YARNS; MECHANICAL FINISHING OF YARNS OR ROPES; WARPING OR BEAMING
    • D02G—CRIMPING OR CURLING FIBRES, FILAMENTS, THREADS, OR YARNS; YARNS OR THREADS
    • D02G3/00—Yarns or threads, e.g. fancy yarns; Processes or apparatus for the production thereof, not otherwise provided for
    • D02G3/02—Yarns or threads characterised by the material or by the materials from which they are made
    • D02G3/04—Blended or other yarns or threads containing components made from different materials
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M11/00—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising
    • D06M11/01—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with hydrogen, water or heavy water; with hydrides of metals or complexes thereof; with boranes, diboranes, silanes, disilanes, phosphines, diphosphines, stibines, distibines, arsines, or diarsines or complexes thereof
    • D06M11/05—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with hydrogen, water or heavy water; with hydrides of metals or complexes thereof; with boranes, diboranes, silanes, disilanes, phosphines, diphosphines, stibines, distibines, arsines, or diarsines or complexes thereof with water, e.g. steam; with heavy water

Definitions

  • the present invention relates to a composite yarn, a method for producing the same, and a fabric.
  • Non-Patent Document 1 Since protein fibers have advantages such as biodegradability and low energy for production and processing, demand for various fields is expected to increase in accordance with the growing awareness of environmental conservation in recent years. I have. Animal fiber such as wool among protein fibers has properties such as warm in winter, cool in summer, high moisture absorption and desorption, and elasticity. (For example, Non-Patent Document 1).
  • animal hair fibers are less flammable than cellulose fibers such as cotton, they have the disadvantage that they are more flammable than synthetic fibers such as polyester fibers.
  • synthetic fibers such as polyester fibers.
  • the present inventors have found that the modified fibroin has excellent combustion resistance.
  • the present invention is based on this new finding.
  • the present invention relates to, for example, the following inventions.
  • [1] Contains artificial fibroin fiber and animal hair fiber, The composite yarn, wherein the artificial fibroin fiber contains a modified fibroin and has a contraction history of being irreversibly contracted by contact with water after spinning.
  • [2] The composite yarn according to [1], wherein the artificial fibroin fiber has a wet shrinkage defined by the following formula of 2% or more.
  • wet shrinkage ⁇ 1 ⁇ (length of artificial fibroin fiber wetted by contact with water / length of artificial fibroin fibril after spinning and before contact with water) ⁇ ⁇ 100 (%) [3]
  • the composite yarn according to [1] or [2], wherein the artificial fibroin fiber has a dry shrinkage defined by the following formula of more than 7%. Drying shrinkage ⁇ 1 ⁇ (length of dried artificial fibroin fiber / length of artificial fibroin raw fiber after spinning and before contact with water) ⁇ ⁇ 100 (%) [4]
  • the composite yarn and the fabric according to the present invention contain the modified fibroin fiber, they are also excellent in moisture absorption and heat generation.
  • the composite yarn according to the present embodiment contains artificial fibroin fiber and animal hair fiber.
  • the composite yarns include, for example, blended yarns, blended yarns, blended yarns, interwoven yarns, plied yarns, covering yarns, and the like.
  • the artificial fibroin fiber according to the present embodiment may contain modified fibroin and have a shrinkage history irreversibly shrunk by contact with water after spinning.
  • the modified fibroin has a domain sequence represented by Formula 1: [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif. Including proteins.
  • the modified fibroin may further have an amino acid sequence (N-terminal sequence and C-terminal sequence) added to one or both of the N-terminal side and the C-terminal side of the domain sequence.
  • the N-terminal sequence and the C-terminal sequence are, but not limited to, typically a region having no repeat of the amino acid motif characteristic of fibroin, and are composed of about 100 amino acids.
  • modified fibroin means artificially produced fibroin (artificial fibroin).
  • the modified fibroin may be a fibroin whose domain sequence is different from the amino acid sequence of naturally occurring fibroin, or may be the same as the amino acid sequence of naturally occurring fibroin.
  • naturally-derived fibroin as used herein is also represented by Formula 1: [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif. Is a protein containing the domain sequence to be determined.
  • Modified fibroin may be a directly used amino acid sequence of a naturally occurring fibroin, or a modified amino acid sequence based on the amino acid sequence of a naturally occurring fibroin (for example, cloned naturally occurring fibroin).
  • the amino acid sequence may be modified by modifying the gene sequence of fibroin), or may be artificially designed and synthesized without using naturally occurring fibroin (for example, a nucleic acid encoding the designed amino acid sequence may be used). Which have a desired amino acid sequence by chemical synthesis).
  • domain sequence refers to a crystalline region unique to fibroin (typically, corresponding to the (A) n motif of the amino acid sequence) and an amorphous region (typically, the REP of the amino acid sequence).
  • the (A) n motif indicates an amino acid sequence mainly containing an alanine residue, and has 2 to 27 amino acid residues.
  • the number of amino acid residues in the n motif may be an integer of 2 to 20, 4 to 27, 4 to 20, 8 to 20, 10 to 20, 4 to 16, 8 to 16, or 10 to 16 .
  • the ratio of the number of alanine residues to the total number of amino acid residues in the n motif may be 40% or more, and is 60% or more, 70% or more, 80% or more, 83% or more, 85% or more, It may be 86% or more, 90% or more, 95% or more, or 100% (meaning that it is composed of only alanine residues).
  • At least seven of the (A) n motifs present in the domain sequence may be composed of only alanine residues.
  • REP indicates an amino acid sequence composed of 2 to 200 amino acid residues.
  • REP may be an amino acid sequence composed of 10 to 200 amino acid residues.
  • m represents an integer of 2 to 300, and may be an integer of 10 to 300.
  • the plurality of (A) n motifs may have the same amino acid sequence or different amino acid sequences.
  • a plurality of REPs may have the same amino acid sequence or different amino acid sequences.
  • the modified fibroin according to the present embodiment has, for example, an amino acid sequence corresponding to, for example, substitution, deletion, insertion and / or addition of one or more amino acid residues with respect to a cloned natural fibroin gene sequence.
  • an amino acid sequence corresponding to, for example, substitution, deletion, insertion and / or addition of one or more amino acid residues with respect to a cloned natural fibroin gene sequence can be obtained by performing the following modification.
  • Substitution, deletion, insertion and / or addition of amino acid residues can be performed by methods well known to those skilled in the art, such as partial specific mutagenesis. Specifically, Nucleic Acid Res. 10, 6487 (1982) and Methods ⁇ in ⁇ Enzymology, 100, 448 (1983).
  • Naturally occurring fibroin is a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif. Yes, specifically, for example, fibroin produced by insects or spiders.
  • fibroin produced by insects examples include Bombyx @ mori, Bombyx @ mandarina, natural silkworm (Antheraea @ yamamai), tussah (Anterea @ pernii), and maple silkworm (Erioganyerii). ), Silkworms produced by silkworms (Samia cynthia), chestnut worms (Caligura japonica), tussah silkworms (Antheraea mylitta), silkworms such as moga silkworms (Antheraea assama), and silkworms produced by larvae of the hornet beetle Hornet silk protein.
  • fibroin produced by insects include, for example, silkworm fibroin L chain (GenBank Accession No. M76430 (base sequence) and AAA27840.1 (amino acid sequence)).
  • Examples of the fibroin produced by spiders include spiders belonging to the genus Araneus (genus Araneus), such as Orion spider, Elder spider, Red-colored Spider, Blue-colored Spider, etc. Spiders belonging to the genus Argiope genus (Genus Pronus), such as spiders belonging to the genus Procarpus spp., Spiders belonging to the genus Pronus, and spider spiders belonging to the genus Cynotarachne, such as the genus Cyrtarachne, such as Torinofundamashi and Otorinofundamashi.
  • Genus Araneus such as Orion spider, Elder spider, Red-colored Spider, Blue-colored Spider, etc.
  • Spiders belonging to the genus Argiope genus such as spiders belonging to the genus Procarpus spp.
  • Spiders belonging to the genus Pronus and spider spiders belonging to the genus Cynotarachne, such as the genus Cyrtarachne, such
  • Spiders belonging to the genus Ordgarius such as spiders belonging to the genus (Gasteracantha), spiders belonging to the genus Orbalis, and spiders belonging to the genus Ordgarius, such as the spiders belonging to the genus Ordgarius.
  • Spiders belonging to the genus Argiope such as Argiope bruennichi, spiders belonging to the genus Argiope sp.
  • Spiders belonging to the genus Spider such as spiders belonging to the genus (Cytophora) and spiders belonging to the genus (Potys), spiders belonging to the genus Spiders (genus Cyclosa) such as the spiders belonging to the genus Cyclosa and spiders belonging to the genus Cygnus spp.
  • Spider silk proteins produced by spiders belonging to the genus Chorizopes), and asina such as red-headed spiders, red-backed spiders, blue-backed spiders and urocore-red spiders Spiders belonging to the genus Tetragnatha, spiders belonging to the genus Tetragnatha, spiders belonging to the genus Leucaegium, such as the spiders Argiope bruennichi and spiders spiders belonging to the genus Leucauge, such as the spiders belonging to the genus Nephila sp.
  • Spiders belonging to the genus Dyschiriognatha such as spiders belonging to the genus Menosira and spiders belonging to the genus Dyschiriognatha, such as the spiders belonging to the genus Latus and the spiders belonging to the genus Lastroconidae belonging to the genus Latus sp.
  • Spiders belonging to the family Tetragnathidae such as spiders belonging to the genus Prostenops (Euprosthenops) are produced.
  • Spider silk protein examples include dragline proteins such as MaSp (MaSp1 and MaSp2) and ADF (ADF3 and ADF4), and MiSp (MiSp1 and MiSp2).
  • spider silk proteins produced by spiders include, for example, fibroin-3 (adf-3) [derived from Araneus diadematus] (GenBank accession number AAC47010 (amino acid sequence), U47855 (base sequence)), fibroin-4 (adf-4) [derived from Araneus diadematus] (GenBank accession number AAC47011 (amino acid sequence), U47856 (base sequence)), dragline silk protein spidroin 1 [derived from amino acid sequence of Nephila claviBAC04A4 and derived from amino acid sequence of ph ), U37520 (base sequence)), major ⁇ ampullate ⁇ spidro n 1 [Derived from Latrodictus hesperus] (GenBank accession number ABR68856 (amino acid sequence), EF595246 (base sequence)), dragline silk protein spidroin 2 [Derived from Nephila clavata (GenBank accession number
  • CAJ00428 amino acid sequence
  • AJ97155 base sequence
  • major ⁇ sample ⁇ spidroin ⁇ 2 [Euprosus] ] GenBank Accession No. CAM32249.1 (amino acid sequence), AM490169 (base sequence)
  • minor ⁇ silk ⁇ protein ⁇ 1 [Nephila ⁇ clavipes] GenBank Accession No. AAC14589.1 (amino acid sequence)
  • minor ⁇ ampilatte [in] Nephila clavipes] GenBank Accession No. AAC14591.1 (amino acid sequence)
  • minor ampoulate spidroin-like protein [Nephilengys Cruenata] GenBank Accession No. ABR3727.1.
  • fibroin in which sequence information is registered in NCBI GenBank.
  • sequence information is registered in NCBI GenBank.
  • spidroin, ampullate, fibroin, "silk and polypeptide", or “silk and protein” is described as a keyword in DEFINITION from among sequences including INV as DIVISION in the sequence information registered in NCBI @ GenBank. It can be confirmed by extracting a character string of a specific product from a sequence or CDS, and extracting a sequence in which a specific character string is described in TISSUE @ TYPE from SOURCE.
  • the modified fibroin according to this embodiment may be a modified silk (silk) fibroin (an amino acid sequence of a silk protein produced by a silkworm modified), and a modified spider silk fibroin (a spider silk protein produced by an arachnid). Modified amino acid sequence).
  • a modified spider silk fibroin is preferable.
  • the modified fibroin include a modified fibroin (first modified fibroin) derived from a large spinal cord marker protein produced in a spider's large ampullate gland, and a domain sequence having a reduced content of glycine residues.
  • first modified fibroin derived from a large spinal cord marker protein produced in a spider's large ampullate gland
  • domain sequence having a reduced content of glycine residues derived from a large spinal cord marker protein produced in a spider's large ampullate gland
  • a second modified fibroin derived from a large spinal cord marker protein produced in a spider's large ampullate gland
  • a second modified fibroin derived from a large spinal cord marker protein produced in a spider's large ampullate gland
  • second modified fibroin derived from a large spinal cord marker protein produced in a spider's large ampullate gland
  • a domain sequence having a reduced content of glycine residues derived from a large spinal cord marker protein produced in a spider's large ampul
  • the first modified fibroin includes a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the number of amino acid residues of the (A) n motif is preferably an integer of 3 to 20, more preferably an integer of 4 to 20, still more preferably an integer of 8 to 20, and an integer of 10 to 20 Is still more preferable, an integer of 4 to 16 is still more preferable, an integer of 8 to 16 is particularly preferable, and an integer of 10 to 16 is most preferable.
  • the number of amino acid residues constituting REP in Formula 1 is preferably 10 to 200 residues, more preferably 10 to 150 residues, and more preferably 20 to 100 residues.
  • the first modified fibroin has a total number of glycine, serine and alanine residues contained in the amino acid sequence represented by Formula 1: [(A) n motif-REP] m
  • the total number is preferably 40% or more, more preferably 60% or more, and even more preferably 70% or more.
  • the first modified fibroin comprises a unit of the amino acid sequence represented by Formula 1: [(A) n motif-REP] m , and has a C-terminal sequence represented by any of SEQ ID NOS: 1 to 3 or
  • the polypeptide may be an amino acid sequence having 90% or more homology with the amino acid sequence shown in any one of SEQ ID NOs: 1 to 3.
  • the amino acid sequence shown in SEQ ID NO: 1 is the same as the amino acid sequence consisting of 50 amino acids at the C-terminal of the amino acid sequence of ADF3 (GI: 1263287, NCBI), and the amino acid sequence shown in SEQ ID NO: 2 is
  • the amino acid sequence shown in SEQ ID NO: 3 is identical to the amino acid sequence shown in SEQ ID NO: 1 by removing 20 residues, and the amino acid sequence shown in SEQ ID NO: 3 is obtained by removing 29 residues from the C-terminal of the amino acid sequence shown in SEQ ID NO: 1. It is identical to the amino acid sequence.
  • the first modified fibroin (1-i) an amino acid sequence represented by SEQ ID NO: 4 (recombinant ⁇ spider ⁇ silk ⁇ protein ⁇ ADF3KaiLargeNRSH1) or (1-ii) an amino acid sequence represented by SEQ ID NO: 4 and 90 Modified fibroin comprising an amino acid sequence having at least% sequence identity.
  • the sequence identity is preferably 95% or more.
  • the amino acid sequence represented by SEQ ID NO: 4 is the same as the amino acid sequence of ADF3 in which an amino acid sequence (SEQ ID NO: 5) comprising an initiation codon, a His10 tag, and an HRV3C protease (Human ⁇ rhinovirus @ 3C protease) recognition site at the N-terminus is added.
  • the 13th repeat region was increased so as to be approximately doubled, and the mutation was mutated so that translation was terminated at the 1154th amino acid residue.
  • the amino acid sequence at the C-terminus of the amino acid sequence represented by SEQ ID NO: 4 is the same as the amino acid sequence represented by SEQ ID NO: 3.
  • the modified fibroin of (1-i) may have an amino acid sequence represented by SEQ ID NO: 4.
  • the second modified fibroin has an amino acid sequence whose domain sequence has a reduced content of glycine residues as compared to naturally occurring fibroin.
  • the second modified fibroin can be said to have an amino acid sequence corresponding to at least one or more glycine residues in the REP replaced by another amino acid residue, as compared to a naturally occurring fibroin. .
  • the second modified fibroin has a domain sequence of GGX and GPGXX in REP (where G is a glycine residue, P is a proline residue, and X is an amino acid residue other than glycine, as compared with a naturally-derived fibroin. At least one motif sequence selected from the group consisting of at least one glycine residue in one or more of the motif sequences has been replaced with another amino acid residue. You may.
  • the ratio of the motif sequence in which the glycine residue is replaced with another amino acid residue may be 10% or more of the entire motif sequence.
  • the second modified fibroin comprises a domain sequence represented by Formula 1: [(A) n motif-REP] m , and from the (A) n motif located at the most C-terminal side to the domain sequence
  • the total number of amino acid residues in the amino acid sequence consisting of XGX (where X represents an amino acid residue other than glycine) contained in all REPs in the sequence excluding the sequence up to the C-terminus is represented by z, From, when the total number of amino acid residues in the sequence excluding the sequence from the (A) n motif located at the most C-terminal side to the C-terminal of the domain sequence is defined as w, z / w is 30% or more; It may have an amino acid sequence of 40% or more, 50% or more, or 50.9% or more.
  • the number of alanine residues relative to the total number of amino acid residues in the n motif may be 83% or more, preferably 86% or more, more preferably 90% or more, and more preferably 95% or more. More preferably, it is even more preferably 100% (meaning that it is composed of only alanine residues).
  • the second modified fibroin is preferably one in which the content of the amino acid sequence consisting of XGX is increased by substituting one glycine residue of the GGX motif with another amino acid residue.
  • the content ratio of the amino acid sequence consisting of GGX in the domain sequence is preferably 30% or less, more preferably 20% or less, further preferably 10% or less, and 6% or less. %, Still more preferably 4% or less, further preferably 2% or less.
  • the content ratio of the amino acid sequence consisting of GGX in the domain sequence can be calculated by the same method as the method for calculating the content ratio (z / w) of the amino acid sequence consisting of XGGX below.
  • z / w (%) can be calculated by dividing z by w.
  • z / w is preferably 50.9% or more, more preferably 56.1% or more, still more preferably 58.7% or more, and 70% or more. Is still more preferred, and even more preferably 80% or more.
  • the upper limit of z / w is not particularly limited, but may be, for example, 95% or less.
  • the second modified fibroin is modified, for example, by replacing at least a part of the base sequence encoding a glycine residue from the cloned natural fibroin gene sequence to encode another amino acid residue.
  • a GGX motif and one glycine residue in the GPGXX motif may be selected, or the glycine residue may be substituted so that z / w becomes 50.9% or more.
  • the amino acid sequence can be obtained by designing an amino acid sequence satisfying the above aspect from the amino acid sequence of naturally occurring fibroin, and chemically synthesizing a nucleic acid encoding the designed amino acid sequence.
  • one or more amino acid residues are further substituted or deleted.
  • the amino acid sequence corresponding to insertion, addition, and / or addition may be modified.
  • the other amino acid residue is not particularly limited as long as it is an amino acid residue other than a glycine residue, but includes a valine (V) residue, a leucine (L) residue, an isoleucine (I) residue, and a methionine ( M) residue, hydrophobic amino acid residue such as proline (P) residue, phenylalanine (F) residue and tryptophan (W) residue, glutamine (Q) residue, asparagine (N) residue, serine (S ) Residues, lysine (K) residues and hydrophilic amino acid residues such as glutamic acid (E) residues, and valine (V) residues, leucine (L) residues, isoleucine (I) residues, phenylalanine ( F) residues and glutamine (Q) residues are more preferred, and glutamine (Q) residues are even more preferred.
  • the second modified fibroin examples include (2-i) SEQ ID NO: 6 (Met-PRT380), SEQ ID NO: 7 (Met-PRT410), SEQ ID NO: 8 (Met-PRT525) or SEQ ID NO: 9 (Met -PRT799), or (2-ii) an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, Modified fibroin can be mentioned.
  • the modified fibroin (2-i) will be described.
  • the amino acid sequence represented by SEQ ID NO: 6 is obtained by replacing all GGX in the REP of the amino acid sequence represented by SEQ ID NO: 10 (Met-PRT313) corresponding to naturally occurring fibroin with GQX.
  • the amino acid sequence represented by SEQ ID NO: 7 is obtained by deleting every two (A) n motifs from the N-terminal side to the C-terminal side from the amino acid sequence represented by SEQ ID NO: 6, and further before the C-terminal sequence. In which one [(A) n motif-REP] was inserted.
  • the amino acid sequence represented by SEQ ID NO: 8 has two alanine residues inserted at the C-terminal side of each (A) n motif of the amino acid sequence represented by SEQ ID NO: 7, and further has a partial glutamine (Q) residue. It has been replaced with a serine (S) residue, and some amino acids on the C-terminal side have been deleted so that the molecular weight becomes almost the same as that of SEQ ID NO: 7.
  • the amino acid sequence represented by SEQ ID NO: 9 has a region of 20 domain sequences present in the amino acid sequence represented by SEQ ID NO: 7 (however, several amino acid residues on the C-terminal side of the region are substituted). Is repeated four times with a predetermined hinge sequence and His tag sequence added to the C-terminal.
  • the value of z / w in the amino acid sequence represented by SEQ ID NO: 10 (corresponding to naturally occurring fibroin) is 46.8%.
  • the values of z / w in the amino acid sequence represented by SEQ ID NO: 6, the amino acid sequence represented by SEQ ID NO: 7, the amino acid sequence represented by SEQ ID NO: 8, and the amino acid sequence represented by SEQ ID NO: 9 are 58.7%, respectively. 70.1%, 66.1% and 70.0%.
  • the value of x / y at the jagged ratio (described later) of 1: 1.8 to 11.3 of the amino acid sequences represented by SEQ ID NO: 10, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9 is as follows: They are 15.0%, 15.0%, 93.4%, 92.7% and 89.8%, respectively.
  • the modified fibroin of (2-i) may have an amino acid sequence represented by SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.
  • the modified fibroin of (2-ii) contains an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9.
  • the modified fibroin of (2-ii) is also a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (2-ii) has 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, and contains XGX ( Where X represents an amino acid residue other than glycine.)
  • z is the total number of amino acid residues in the amino acid sequence consisting of Is preferably 50.9% or more.
  • the second modified fibroin may include a tag sequence at one or both of the N-terminus and the C-terminus. As a result, the modified fibroin can be isolated, immobilized, detected, visualized, and the like.
  • the tag sequence examples include an affinity tag utilizing specific affinity (binding property, affinity) with another molecule.
  • affinity tag is a histidine tag (His tag).
  • His tag is a short peptide in which about 4 to 10 histidine residues are arranged, and has a property of specifically binding to a metal ion such as nickel. Therefore, isolation of a modified fibroin by metal chelation chromatography (chelating @ metal @ chromatography).
  • SEQ ID NO: 11 amino acid sequence including a His tag sequence and a hinge sequence.
  • tag sequences such as glutathione-S-transferase (GST), which specifically binds to glutathione, and maltose binding protein (MBP), which specifically binds to maltose, can be used.
  • GST glutathione-S-transferase
  • MBP maltose binding protein
  • an “epitope tag” utilizing an antigen-antibody reaction can be used.
  • a peptide (epitope) showing antigenicity as a tag sequence an antibody against the epitope can be bound.
  • the epitope tag include an HA (peptide sequence of hemagglutinin of influenza virus) tag, myc tag, and FLAG tag.
  • a tag sequence that can be cleaved by a specific protease can be used.
  • protease treatment By subjecting the protein adsorbed via the tag sequence to protease treatment, the modified fibroin from which the tag sequence has been separated can also be recovered.
  • modified fibroin containing a tag sequence (2-iii) the amino acid represented by SEQ ID NO: 12 (PRT380), SEQ ID NO: 13 (PRT410), SEQ ID NO: 14 (PRT525), or SEQ ID NO: 15 (PRT799)
  • Modified fibroin comprising a sequence or (2-iv) an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15 can be mentioned. .
  • amino acid sequences represented by SEQ ID NO: 16 (PRT313), SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15 are represented by SEQ ID NO: 10, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9, respectively.
  • An amino acid sequence represented by SEQ ID NO: 11 (including a His tag sequence and a hinge sequence) is added to the N-terminal of the amino acid sequence shown.
  • the modified fibroin of (2-iii) may have an amino acid sequence represented by SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15.
  • the modified fibroin of (2-iv) includes an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15.
  • the modified fibroin of (2-iv) is also a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (2-iv) has 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15, and contains XGX (where X represents an amino acid residue other than glycine.)
  • z is the total number of amino acid residues in the amino acid sequence consisting of Is preferably 50.9% or more.
  • the second modified fibroin may include a secretion signal for releasing a protein produced in the recombinant protein production system to the outside of the host.
  • the sequence of the secretion signal can be appropriately set according to the type of the host.
  • the third modified fibroin has an amino acid sequence whose domain sequence has a reduced content of the (A) n motif as compared to a naturally occurring fibroin. It can be said that the domain sequence of the third modified fibroin has an amino acid sequence corresponding to the deletion of at least one or a plurality of (A) n motifs as compared to naturally occurring fibroin.
  • the third modified fibroin may have an amino acid sequence corresponding to 10 to 40% deletion of the (A) n motif from naturally occurring fibroin.
  • the third modification fibroin its domain sequence, compared to the naturally occurring fibroin, at least from the N-terminal side toward the C-terminal one to three (A) n motif every one (A) n motif May have an amino acid sequence corresponding to the deletion of
  • the third modified fibroin has a domain sequence deletion of at least two (A) n motifs from the N-terminal side to the C-terminal side, and one (A) It may have an amino acid sequence corresponding to the deletion of the n motif repeated in this order.
  • the third modified fibroin may have a domain sequence having an amino acid sequence corresponding to the deletion of the (A) n motif at least every third sequence from the N-terminal side to the C-terminal side. .
  • the third modified fibroin comprises a domain sequence represented by Formula 1: [(A) n motif-REP] m , and two adjacent [(A) n motifs from the N-terminal side to the C-terminal side] -REP]
  • the number of amino acid residues of REP in the unit is sequentially compared, and when the number of amino acid residues of REP having a small number of amino acid residues is set to 1, the ratio of the number of amino acid residues of the other REP is 1.8 to When the maximum value of the sum of the number of amino acid residues of two adjacent [(A) n motif-REP] units that is 11.3 is x, and the total number of amino acid residues in the domain sequence is y In addition, it may have an amino acid sequence in which x / y is 20% or more, 30% or more, 40% or more, or 50% or more.
  • the number of alanine residues relative to the total number of amino acid residues in the n motif may be 83% or more, preferably 86% or more, more preferably 90% or more, and more preferably 95% or more. More preferably, it is even more preferably 100% (meaning that it is composed of only alanine residues).
  • FIG. 1 shows a domain sequence obtained by removing the N-terminal sequence and the C-terminal sequence from the modified fibroin. From the N-terminal side (left side), the domain sequence is (A) n motif-first REP (50 amino acid residues)-(A) n motif-second REP (100 amino acid residues)-(A) n Motif-third REP (10 amino acid residues)-(A) n motif-fourth REP (20 amino acid residues)-(A) n motif-fifth REP (30 amino acid residues)-(A) It has a sequence called an n motif.
  • FIG. 1 shows pattern 1 (comparison of the first REP and the second REP, and comparison of the third REP with the fourth REP), pattern 2 (comparison of the first REP and the second REP, and Pattern 4 (comparison of the second REP with the third REP, and comparison of the fourth REP with the fifth REP), pattern 4 (the comparison of the fourth REP with the fifth REP), and pattern 4 (the first REP with the fifth REP). Second REP comparison). Note that there are other selection methods.
  • the number of amino acid residues of each REP in two selected adjacent [(A) n motif-REP] units is compared.
  • each pattern the total number of amino acid residues of two adjacent [(A) n motif-REP] units shown by a solid line is added (not only the REP but also the number of amino acid residues of the (A) n motif. is there.). Then, the sum total is compared, and the total value (maximum value of the total values) of the patterns having the maximum total value is x. In the example shown in FIG. 1, the total value of pattern 1 is the maximum.
  • x / y (%) can be calculated by dividing x by the total number of amino acid residues y in the domain sequence.
  • x / y is preferably at least 50%, more preferably at least 60%, further preferably at least 65%, and even more preferably at least 70%. Preferably, it is still more preferably at least 75%, particularly preferably at least 80%.
  • the upper limit of x / y is not particularly limited, and may be, for example, 100% or less. When the indentation ratio is 1: 1.9 to 11.3, x / y is preferably 89.6% or more, and when the indentation ratio is 1: 1.8 to 3.4, x / y is x / y.
  • / Y is preferably at least 77.1%, and when the jagged ratio is 1: 1.9 to 8.4, x / y is preferably at least 75.9%, and the jagged ratio is 1 In the case of 1.9 to 4.1, x / y is preferably at least 64.2%.
  • x / y should be 46.4% or more. Is preferably 50% or more, more preferably 55% or more, still more preferably 60% or more, even more preferably 70% or more, and even more preferably 80% or more. It is particularly preferred that there is.
  • the upper limit of x / y is not particularly limited, and may be 100% or less.
  • x / y in naturally occurring fibroin will be described.
  • 663 types of fibroins (among them, 415 types of spider-derived fibroins) were extracted.
  • x / y was calculated from the amino acid sequence of the naturally occurring fibroin composed of the domain sequence represented by Formula 1: [(A) n motif-REP] m by the above calculation method.
  • FIG. 3 shows the results when the indentation ratio is 1: 1.9 to 4.1.
  • the horizontal axis in FIG. 3 indicates x / y (%), and the vertical axis indicates frequency.
  • the x / y of the naturally derived fibroin is less than 64.2% (the highest is 64.14%).
  • the third modified fibroin deletes one or more of the sequence encoding the (A) n motif from the cloned natural fibroin gene sequence such that x / y is 64.2% or more. Can be obtained. Further, for example, an amino acid sequence corresponding to deletion of one or more (A) n motifs is designed so that x / y is 64.2% or more based on the amino acid sequence of naturally occurring fibroin. It can also be obtained by chemically synthesizing a nucleic acid encoding the amino acid sequence.
  • amino acid residues are further substituted, deleted, inserted and / or added.
  • Amino acid sequence modification corresponding to the above may be performed.
  • the third modified fibroin (3-i) SEQ ID NO: 17 (Met-PRT399), SEQ ID NO: 7 (Met-PRT410), SEQ ID NO: 8 (Met-PRT525), or SEQ ID NO: 9 (Met-PRT525) -PRT799), or (3-ii) an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 17, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, Modified fibroin can be mentioned.
  • the modified fibroin (3-i) will be described.
  • the amino acid sequence represented by SEQ ID NO: 17 differs from the amino acid sequence represented by SEQ ID NO: 10 (Met-PRT313) corresponding to naturally occurring fibroin in that every two amino acids from the N-terminal side to the C-terminal side (A) n The motif was deleted, and one [(A) n motif-REP] was inserted before the C-terminal sequence.
  • the amino acid sequence represented by SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9 is as described for the second modified fibroin.
  • the amino acid sequence represented by SEQ ID NO: 10 (corresponding to naturally-occurring fibroin) has an x / y value of 15.0% at a giza ratio of 1: 1.8-11.3.
  • the value of x / y in the amino acid sequence represented by SEQ ID NO: 17 and the amino acid sequence represented by SEQ ID NO: 7 is 93.4%.
  • the value of x / y in the amino acid sequence represented by SEQ ID NO: 8 is 92.7%.
  • the value of x / y in the amino acid sequence represented by SEQ ID NO: 9 is 89.8%.
  • the values of z / w in the amino acid sequences represented by SEQ ID NO: 10, SEQ ID NO: 17, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9 are 46.8%, 56.2%, 70.1%, 66. 1% and 70.0%.
  • the modified fibroin of (3-i) may be composed of the amino acid sequence represented by SEQ ID NO: 17, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.
  • the modified fibroin of (3-ii) contains an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 17, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.
  • the modified fibroin of (3-ii) is also a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the sequence identity is preferably 95% or more.
  • the modified fibroin (3-ii) has 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 17, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, and is N-terminal to C-terminal.
  • the number of amino acid residues of REP of two adjacent [(A) n motif-REP] units is sequentially compared, and when the number of amino acid residues of REP having a small number of amino acid residues is set to 1, the other Amino acid residues of two adjacent [(A) n motif-REP] units having a ratio of the number of amino acid residues of REP of 1.8 to 11.3 (a giza ratio of 1: 1.8 to 11.3).
  • x / y be 64.2% or more, where x is the maximum value of the sum of the base numbers and y is the total number of amino acid residues in the domain sequence.
  • the third modified fibroin may include the above-described tag sequence at one or both of the N-terminus and the C-terminus.
  • modified fibroin containing a tag sequence (3-iii) the amino acid represented by SEQ ID NO: 18 (PRT399), SEQ ID NO: 13 (PRT410), SEQ ID NO: 14 (PRT525), or SEQ ID NO: 15 (PRT799)
  • Modified fibroin comprising a sequence or (3-iv) an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 18, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15 can be mentioned. .
  • amino acid sequences represented by SEQ ID NO: 18, SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15 are obtained by adding SEQ ID NO: 11 to the N-terminal of the amino acid sequences represented by SEQ ID NO: 17, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, respectively. (Including a His tag sequence and a hinge sequence).
  • the modified fibroin of (3-iii) may have an amino acid sequence represented by SEQ ID NO: 18, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15.
  • the modified fibroin of (3-iv) includes an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 18, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15.
  • the modified fibroin of (3-iv) is also a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (3-iv) has 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 18, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15 and is N-terminal to C-terminal.
  • the number of amino acid residues of REP of two adjacent [(A) n motif-REP] units is sequentially compared, and when the number of amino acid residues of REP having a small number of amino acid residues is set to 1, the other
  • the maximum value of the total value obtained by adding the number of amino acid residues of two adjacent [(A) n motif-REP] units having a ratio of the number of amino acid residues of REP of 1.8 to 11.3 is defined as x.
  • x / y is 64.2% or more, where y is the total number of amino acid residues in the domain sequence.
  • the third modified fibroin may include a secretion signal for releasing a protein produced in the recombinant protein production system to the outside of the host.
  • the sequence of the secretion signal can be appropriately set according to the type of the host.
  • the fourth modified fibroin has an amino acid sequence whose domain sequence has a reduced content of a glycine residue in addition to the content of the (A) n motif reduced as compared to a naturally-derived fibroin.
  • the domain sequence of the fourth modified fibroin is at least one or more (A) n motifs deleted, and further at least one or more glycine residues in the REP, as compared to naturally occurring fibroin. It can be said that it has an amino acid sequence equivalent to being replaced with another amino acid residue. That is, the fourth modified fibroin is a modified fibroin having both the characteristics of the second modified fibroin and the third modified fibroin described above. Specific aspects and the like are as described for the second modified fibroin and the third modified fibroin.
  • the fourth modified fibroin (4-i) SEQ ID NO: 7 (Met-PRT410), SEQ ID NO: 8 (Met-PRT525), SEQ ID NO: 9 (Met-PRT799), SEQ ID NO: 13 (PRT410) ), The amino acid sequence represented by SEQ ID NO: 14 (PRT525) or SEQ ID NO: 15 (PRT799), or (4-ii) SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15
  • a modified fibroin comprising an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by.
  • Specific embodiments of the modified fibroin comprising the amino acid sequence represented by SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15 are as described above.
  • the fifth modified fibroin has a domain sequence in which one or more amino acid residues in REP have been replaced by amino acid residues having a large hydrophobicity index as compared to naturally occurring fibroin, and / or It may have an amino acid sequence locally including a region having a large hydrophobicity index, corresponding to insertion of one or more amino acid residues having a large hydrophobicity index therein.
  • a region having a locally large hydrophobicity index is preferably composed of 2 to 4 consecutive amino acid residues.
  • the amino acid residue having a large hydrophobicity index is selected from isoleucine (I), valine (V), leucine (L), phenylalanine (F), cysteine (C), methionine (M), and alanine (A). More preferably, it is a residue.
  • the fifth modified fibroin may have one or more amino acid residues in the REP replaced with amino acid residues having a higher hydrophobicity index, and / or one or more amino acids in the REP as compared to a naturally occurring fibroin.
  • one or more amino acid residues may be substituted, deleted, inserted and / or added as compared with naturally occurring fibroin.
  • the fifth modified fibroin is, for example, one or more hydrophilic amino acid residues (for example, amino acid residues having a negative hydrophobicity index) in the REP from the cloned natural fibroin gene sequence, It can be obtained by substituting a group (for example, an amino acid residue having a positive hydrophobicity index) and / or inserting one or more hydrophobic amino acid residues into REP.
  • a group for example, an amino acid residue having a positive hydrophobicity index
  • one or more hydrophilic amino acid residues in REP were replaced with hydrophobic amino acid residues from the amino acid sequence of naturally occurring fibroin, and / or one or more hydrophobic amino acid residues in REP.
  • one or more hydrophilic amino acid residues in REP were replaced with hydrophobic amino acid residues from the amino acid sequence of naturally occurring fibroin, and / or one or more hydrophobic amino acid residues in REP.
  • the amino acid sequence corresponding to the substitution, deletion, insertion and / or addition of one or more amino acid residues may be further modified.
  • the fifth modified fibroin contains a domain sequence represented by Formula 1: [(A) n motif-REP] m and extends from the (A) n motif located at the most C-terminal side to the C-terminus of the domain sequence.
  • the total number of amino acid residues included in a region where the average value of the hydrophobicity index of four consecutive amino acid residues is 2.6 or more is p,
  • q the total number of amino acid residues contained in the sequence excluding the sequence from the (A) n motif located at the most C-terminal side to the C-terminal of the domain sequence from the domain sequence is defined as q
  • p / q is 6 .2% or more.
  • hydrophobicity index of amino acid residues
  • a publicly known index Kyte J, & Doolittle R (1982) "A simple method for display, the hydropathic charactor of aa protein, J.Pol.Mol. 105-132).
  • HI hydropathic index
  • sequence A [(A) n motif-REP] m (Hereinafter, referred to as “sequence A”).
  • sequence A the average value of the hydrophobicity index of four consecutive amino acid residues is calculated. The average value of the hydrophobicity index is determined by dividing the total sum of HI of each amino acid residue contained in four consecutive amino acid residues by 4 (the number of amino acid residues).
  • the average value of the hydrophobicity index is determined for all four consecutive amino acid residues (each amino acid residue is used for calculating the average value one to four times). Next, a region where the average value of the hydrophobicity index of four consecutive amino acid residues is 2.6 or more is specified. Even when a certain amino acid residue corresponds to a plurality of “consecutive four amino acid residues having an average value of the hydrophobicity index of 2.6 or more”, it is included as one amino acid residue in the region. become. Then, the total number of amino acid residues contained in the region is p. The total number of amino acid residues contained in sequence A is q.
  • p / q is preferably 6.2% or more, more preferably 7% or more, further preferably 10% or more, and more preferably 20% or more. Even more preferably, it is even more preferably 30% or more.
  • the upper limit of p / q is not particularly limited, but may be, for example, 45% or less.
  • the fifth modified fibroin is, for example, one or a plurality of hydrophilic amino acid residues (for example, a hydrophobicity index) in the REP so that the amino acid sequence of the cloned natural fibroin is satisfied so as to satisfy the above-mentioned p / q conditions.
  • a hydrophobic amino acid residue eg, an amino acid residue having a positive hydrophobicity index
  • inserting one or more hydrophobic amino acid residues into the REP By doing so, it can be obtained by locally modifying the amino acid sequence to include a region having a large hydrophobicity index.
  • an amino acid sequence satisfying the above-mentioned p / q condition from the amino acid sequence of naturally occurring fibroin and chemically synthesizing a nucleic acid encoding the designed amino acid sequence.
  • one or more amino acid residues in the REP have been replaced by amino acid residues with a higher hydrophobicity index and / or one or more amino acid residues in the REP as compared to naturally occurring fibroin.
  • a modification corresponding to substitution, deletion, insertion, and / or addition of one or more amino acid residues may be performed. .
  • the amino acid residue having a large hydrophobicity index is not particularly limited, but isoleucine (I), valine (V), leucine (L), phenylalanine (F), cysteine (C), methionine (M), and alanine (A Is preferred, and valine (V), leucine (L) and isoleucine (I) are more preferred.
  • the fifth modified fibroin (5-i) the amino acid sequence represented by SEQ ID NO: 19 (Met-PRT665), SEQ ID NO: 20 (Met-PRT665), or SEQ ID NO: 21 (Met-PRT666); Or (5-ii) a modified fibroin comprising an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 19, SEQ ID NO: 20, or SEQ ID NO: 21.
  • the modified fibroin of (5-i) will be described.
  • the amino acid sequence represented by SEQ ID NO: 19 consists of three amino acid residues every other REP, except for the domain sequence of the terminal at the C-terminal side, with respect to the amino acid sequence represented by SEQ ID NO: 7 (Met-PRT410).
  • two amino acid sequences (VLI) were inserted, some glutamine (Q) residues were further substituted with serine (S) residues, and some C-terminal amino acids were deleted.
  • the amino acid sequence represented by SEQ ID NO: 20 is obtained by inserting one amino acid sequence (VLI) consisting of three amino acid residues every other REP into the amino acid sequence represented by SEQ ID NO: 8 (Met-PRT525). is there.
  • the amino acid sequence represented by SEQ ID NO: 21 is obtained by inserting two amino acid sequences (VLI) each consisting of three amino acid residues every other REP into the amino acid sequence represented by SEQ ID NO: 8.
  • the modified fibroin of (5-i) may be composed of the amino acid sequence represented by SEQ ID NO: 19, SEQ ID NO: 20 or SEQ ID NO: 21.
  • the modified fibroin of (5-ii) contains an amino acid sequence having 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 19, SEQ ID NO: 20, or SEQ ID NO: 21.
  • the modified fibroin of (5-ii) is also a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (5-ii) has 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 19, SEQ ID NO: 20, or SEQ ID NO: 21, and is located at the most C-terminal side (A) n
  • amino acids contained in a region where the average value of the hydrophobicity index of four consecutive amino acid residues is 2.6 or more When the total number of residues is p, and the total number of amino acid residues contained in the sequence obtained by removing the sequence from the (A) n motif located at the most C-terminal side to the C-terminal of the domain sequence from the domain sequence is q , P / q is preferably at least 6.2%.
  • the fifth modified fibroin may include a tag sequence at one or both of the N-terminus and the C-terminus.
  • modified fibroin containing a tag sequence (5-iii) the amino acid sequence represented by SEQ ID NO: 22 (PRT720), SEQ ID NO: 23 (PRT665) or SEQ ID NO: 24 (PRT666), or (5-iv) ) Modified fibroin comprising an amino acid sequence having 90% or more sequence identity with the amino acid sequence shown in SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24.
  • amino acid sequences represented by SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24 correspond to the amino acid sequence represented by SEQ ID NO: 11 (His tag) at the N-terminal of the amino acid sequences represented by SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21, respectively. Sequences and hinge sequences).
  • the modified fibroin of (5-iii) may have an amino acid sequence represented by SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24.
  • the modified fibroin of (5-iv) contains an amino acid sequence having 90% or more sequence identity with the amino acid sequence shown in SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24.
  • the modified fibroin of (5-iv) is also a protein containing a domain sequence represented by Formula 1: [(A) n motif-REP] m .
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (5-iv) has 90% or more sequence identity with the amino acid sequence represented by SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24, and is located at the most C-terminal side (A) n
  • all REPs contained in the sequence excluding the sequence from the motif to the C-terminus of the domain sequence from the domain sequence amino acids contained in a region where the average value of the hydrophobicity index of four consecutive amino acid residues is 2.6 or more
  • P / q is preferably at least 6.2%.
  • the fifth modified fibroin may include a secretion signal for releasing a protein produced in the recombinant protein production system to the outside of the host.
  • the sequence of the secretion signal can be appropriately set according to the type of the host.
  • the sixth modified fibroin has an amino acid sequence in which the content of glutamine residues is reduced as compared with naturally occurring fibroin.
  • the sixth modified fibroin preferably contains at least one motif selected from GGX motif and GPGXX motif in the amino acid sequence of REP.
  • the content of the GPGXX motif is usually 1% or more, and may be 5% or more, and preferably 10% or more.
  • the upper limit of the GPGXX motif content is not particularly limited, and may be 50% or less, or 30% or less.
  • the “GPGXX motif content” is a value calculated by the following method.
  • Formula 1 Fibroin containing a domain sequence represented by [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif (modified fibroin or naturally occurring fibroin) Fibroin), the number of GPGXX motifs contained in the region of all REPs contained in the sequence excluding the sequence from the (A) n motif located at the most C-terminal side to the C-terminus of the domain sequence from the domain sequence
  • the number obtained by multiplying the total number by 3 ie, the total number of G and P in the GPGXX motif
  • the sequence from the (A) n motif located at the most C-terminal side to the C-terminal of the domain sequence is represented (A)
  • “the sequence obtained by removing the sequence from the (A) n motif located at the most C-terminal side to the C-terminal of the domain sequence from the domain sequence” to “the most C-terminal side” (A) Sequence from n motif to C-terminus of domain sequence (sequence corresponding to REP) may include a sequence having low correlation with a sequence characteristic of fibroin, and m may be small. In this case (that is, when the domain sequence is short), the calculation result of the GPGXX motif content is affected, so that this effect is eliminated.
  • “GPGXX motif” is located at the C-terminus of the REP, even when “XX” is, for example, “AA”, it is treated as a “GPGXX motif”.
  • FIG. 5 is a schematic diagram showing the domain sequence of a modified fibroin.
  • the method of calculating the content rate of the GPGXX motif will be specifically described with reference to FIG. First, in the domain sequence of the modified fibroin shown in FIG. 5 (“[(A) n motif-REP] m- (A) n motif” type), all REPs are located at the most C-terminal side.
  • (A) A sequence obtained by removing the sequence from the n motif to the C-terminus of the domain sequence from the domain sequence ”(the sequence shown as“ region A ”in FIG. 5).
  • the sixth modified fibroin preferably has a glutamine residue content of 9% or less, more preferably 7% or less, still more preferably 4% or less, and particularly preferably 0%. .
  • the “glutamine residue content” is a value calculated by the following method.
  • Formula 1 Fibroin containing a domain sequence represented by [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif (modified fibroin or naturally occurring fibroin) Fibroin), the sequence (the sequence corresponding to “region A” in FIG. 5) in which the sequence from the (A) n motif located closest to the C-terminal side to the C-terminal of the domain sequence is excluded from the domain sequence.
  • the total number of glutamine residues contained in the region is defined as u, and the sequence from the (A) n motif located at the most C-terminal side to the C-terminal of the domain sequence is excluded from the domain sequence, and further (A) n
  • the glutamine residue content is calculated as u / t, where t is the total number of amino acid residues in all REPs excluding the motif.
  • the reason why the “sequence in which the sequence from the (A) n motif located closest to the C-terminal to the C-terminal of the domain sequence is excluded from the domain sequence” is targeted is as described above. The same is true.
  • the sixth modified fibroin corresponds to the fact that its domain sequence has one or more glutamine residues in the REP deleted or replaced with other amino acid residues, as compared to the naturally occurring fibroin. It may have an amino acid sequence.
  • the “other amino acid residue” may be an amino acid residue other than a glutamine residue, but is preferably an amino acid residue having a larger hydrophobicity index than a glutamine residue.
  • the hydrophobicity index of amino acid residues is as shown in Table 1.
  • amino acid residues having a larger hydrophobicity index than glutamine residues include isoleucine (I), valine (V), leucine (L), phenylalanine (F), cysteine (C), and methionine (M )
  • Amino acid residues selected from alanine (A), glycine (G), threonine (T), serine (S), tryptophan (W), tyrosine (Y), proline (P) and histidine (H). it can.
  • an amino acid residue selected from isoleucine (I), valine (V), leucine (L), phenylalanine (F), cysteine (C), methionine (M) and alanine (A) is more preferable.
  • the sixth modified fibroin preferably has a hydrophobicity of REP of -0.8 or more, more preferably -0.7 or more, still more preferably 0 or more, and 0.3 or more. Is still more preferable, and it is particularly preferable that it is 0.4 or more.
  • the upper limit of the hydrophobicity of REP is not particularly limited, and may be 1.0 or less, or may be 0.7 or less.
  • “REP hydrophobicity” is a value calculated by the following method.
  • Formula 1 Fibroin containing a domain sequence represented by [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif (modified fibroin or naturally occurring fibroin) Fibroin), the sequence (the sequence corresponding to “region A” in FIG. 5) in which the sequence from the (A) n motif located closest to the C-terminal side to the C-terminal of the domain sequence is excluded from the domain sequence.
  • the sum of the hydrophobicity indices of each amino acid residue in the region is defined as v, and the sequence from the (A) n motif located closest to the C-terminal side to the C-terminal of the domain sequence is removed from the domain sequence.
  • A) The hydrophobicity of REP is calculated as v / t, where t is the total number of amino acid residues of all REPs excluding n motifs.
  • the degree of hydrophobicity of the REP the reason why the “sequence in which the sequence from the (A) n motif located closest to the C-terminal side to the C-terminal of the domain sequence is excluded from the domain sequence” is targeted is as follows. The same is true.
  • the sixth modified fibroin may have a domain sequence that is missing one or more glutamine residues in the REP and / or one or more glutamine residues in the REP, as compared to the naturally occurring fibroin.
  • the sixth modified fibroin may, for example, delete one or more glutamine residues in the REP from the cloned naturally occurring fibroin gene sequence and / or remove one or more glutamine residues in the REP. By substituting the amino acid residue with, for example, one or more glutamine residues in REP were deleted from the amino acid sequence of naturally occurring fibroin, and / or one or more glutamine residues in REP were replaced with other amino acid residues. It can also be obtained by designing an amino acid sequence corresponding to the above and chemically synthesizing a nucleic acid encoding the designed amino acid sequence.
  • SEQ ID NO: 25 (Met-PRT988), SEQ ID NO: 26 (Met-PRT965), SEQ ID NO: 27 (Met-PRT889), SEQ ID NO: 28 (Met-PRT889) -PRT916), SEQ ID NO: 29 (Met-PRT918), SEQ ID NO: 30 (Met-PRT699), SEQ ID NO: 31 (Met-PRT698), SEQ ID NO: 32 (Met-PRT966), SEQ ID NO: 41 (Met-PRT917) or sequence No.
  • modified fibroin comprising the amino acid sequence represented by (6-ii) SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31
  • An amino acid represented by SEQ ID NO: 32, SEQ ID NO: 41 or SEQ ID NO: 42 It can be mentioned modified fibroin comprising an amino acid sequence having a sequence at least 90% sequence identity.
  • the modified fibroin of (6-i) will be described.
  • the amino acid sequence represented by SEQ ID NO: 25 is obtained by substituting VL for all QQ in the amino acid sequence represented by SEQ ID NO: 7 (Met-PRT410).
  • the amino acid sequence represented by SEQ ID NO: 26 is obtained by substituting all QQs in the amino acid sequence represented by SEQ ID NO: 7 with TS, and substituting the remaining Q with A.
  • the amino acid sequence represented by SEQ ID NO: 27 is obtained by substituting all QQs in the amino acid sequence represented by SEQ ID NO: 7 with VL, and substituting the remaining Q with I.
  • the amino acid sequence represented by SEQ ID NO: 28 is obtained by substituting all QQ in the amino acid sequence represented by SEQ ID NO: 7 with VI and substituting the remaining Q with L.
  • the amino acid sequence represented by SEQ ID NO: 29 is obtained by substituting all QQs in the amino acid sequence represented by SEQ ID NO: 7 with VF and substituting the remaining Q with I.
  • amino acid sequence represented by SEQ ID NO: 30 is obtained by replacing all QQ in the amino acid sequence represented by SEQ ID NO: 8 (Met-PRT525) with VL.
  • the amino acid sequence represented by SEQ ID NO: 31 is obtained by substituting all QQs in the amino acid sequence represented by SEQ ID NO: 8 with VL and substituting the remaining Q with I.
  • the amino acid sequence represented by SEQ ID NO: 32 replaces all QQs in the sequence obtained by repeating twice the domain of the 20 domain sequences present in the amino acid sequence represented by SEQ ID NO: 7 (Met-PRT410) with VF, In addition, the remaining Q is replaced with I.
  • the amino acid sequence represented by SEQ ID NO: 41 (Met-PRT917) is obtained by substituting all QQ in the amino acid sequence represented by SEQ ID NO: 7 with LI and replacing the remaining Q with V.
  • the amino acid sequence represented by SEQ ID NO: 42 (Met-PRT1028) is obtained by substituting all QQ in the amino acid sequence represented by SEQ ID NO: 7 with IF, and substituting the remaining Q with T.
  • amino acid sequences represented by SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 41 and SEQ ID NO: 42 all have glutamine residues.
  • the group content is 9% or less (Table 2).
  • the modified fibroin of (6-i) has SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 41 or SEQ ID NO: It may consist of the amino acid sequence shown.
  • the modified fibroin of (6-ii) has SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 41 or SEQ ID NO: It contains an amino acid sequence having 90% or more sequence identity with the amino acid sequence shown.
  • the modified fibroin of (6-ii) also has a domain represented by Formula 1: [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif A protein containing a sequence.
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (6-ii) preferably has a glutamine residue content of 9% or less. Further, the modified fibroin of (6-ii) preferably has a GPGXX motif content of 10% or more.
  • the sixth modified fibroin may include a tag sequence at one or both of the N-terminus and the C-terminus. As a result, the modified fibroin can be isolated, immobilized, detected, visualized, and the like.
  • modified fibroin containing the tag sequence (6-iii) SEQ ID NO: 33 (PRT888), SEQ ID NO: 34 (PRT965), SEQ ID NO: 35 (PRT889), SEQ ID NO: 36 (PRT916), SEQ ID NO: 37 (PRT918), SEQ ID NO: 38 (PRT699), SEQ ID NO: 39 (PRT698), SEQ ID NO: 40 (PRT966), SEQ ID NO: 43 (PRT917) or modified fibroin comprising the amino acid sequence represented by SEQ ID NO: 44 (PRT1028), or ( 6-iv) The amino acid sequence represented by SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 43 or SEQ ID NO: 44 and 90 % Modified amino acid sequence having an amino acid sequence having at least Mention may be
  • amino acid sequences represented by SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 43, and SEQ ID NO: 44 correspond to SEQ ID NO: 25, respectively.
  • SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39 , SEQ ID NO: 40, SEQ ID NO: 43 and SEQ ID NO: 44 all have a glutamine residue content of 9% or less (Table 3).
  • the modified fibroin of (6-iii) has SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 43 or SEQ ID NO: 44 It may consist of the amino acid sequence shown.
  • the modified fibroin of (6-iv) has SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 43 or SEQ ID NO: 44 It contains an amino acid sequence having 90% or more sequence identity with the amino acid sequence shown.
  • the modified fibroin of (6-iv) also has a domain represented by Formula 1: [(A) n motif-REP] m or Formula 2: [(A) n motif-REP] m- (A) n motif A protein containing a sequence.
  • the sequence identity is preferably 95% or more.
  • the modified fibroin of (6-iv) preferably has a glutamine residue content of 9% or less.
  • the modified fibroin of (6-iv) preferably has a GPGXX motif content of 10% or more.
  • the sixth modified fibroin may contain a secretion signal for releasing a protein produced in the recombinant protein production system to the outside of the host.
  • the sequence of the secretion signal can be appropriately set according to the type of the host.
  • the modified fibroin is at least two or more of the characteristics of the first modified fibroin, the second modified fibroin, the third modified fibroin, the fourth modified fibroin, the fifth modified fibroin, and the sixth modified fibroin. Modified fibroin having both of the following characteristics may be used.
  • the modified fibroin may be a hydrophilic modified fibroin or a hydrophobic modified fibroin.
  • hydrophobic modified fibroin refers to a value obtained by calculating the sum of the hydrophobicity indexes (HI) of all amino acid residues constituting the modified fibroin, and then dividing the total by the total number of amino acid residues. Modified fibroin having an (average HI) greater than 0. The hydrophobicity index is as shown in Table 1. Further, “hydrophilic modified fibroin” is a modified fibroin having an average HI of 0 or less. As the modified fibroin, a hydrophilic modified fibroin is preferred from the viewpoint of excellent combustion resistance, and a hydrophobic modified fibroin is preferred from the viewpoint of excellent moisture absorption and heat generation.
  • hydrophobically modified fibroin examples include the amino acid sequence represented by SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 or SEQ ID NO: 43, SEQ ID NO: 35, Modified fibroin comprising the amino acid sequence represented by SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 or SEQ ID NO: 44.
  • hydrophilic modified fibroin examples include the amino acid sequence represented by SEQ ID NO: 4, the amino acid sequence represented by SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 11, SEQ ID NO: Or the amino acid sequence represented by SEQ ID NO: 15, the amino acid sequence represented by SEQ ID NO: 18, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9, the amino acid sequence represented by SEQ ID NO: 17, SEQ ID NO: 11, SEQ ID NO: 14, or SEQ ID NO: 15 Modified fibroin comprising the amino acid sequence represented by the sequence, SEQ ID NO: 19, SEQ ID NO: 20, or SEQ ID NO: 21.
  • the artificial fibroin fiber according to the present embodiment may contain one kind of modified fibroin alone or may contain two or more kinds of modified fibroin in combination.
  • the modified fibroin may be, for example, a host transformed with an expression vector having a nucleic acid sequence encoding the modified fibroin and one or more regulatory sequences operably linked to the nucleic acid sequence. Can be produced by expressing the nucleic acid.
  • the method for producing the nucleic acid encoding the modified fibroin is not particularly limited.
  • the nucleic acid is produced by a method of amplifying and cloning by a polymerase chain reaction (PCR) or the like using a gene encoding a natural fibroin and modifying it by a genetic engineering technique, or a chemical synthesis method. can do.
  • the method for chemically synthesizing nucleic acids is not particularly limited.
  • AKTA oligopilot plus10010 / 100 Genes can be chemically synthesized by a method of linking oligonucleotides synthesized automatically by PCR or the like.
  • a nucleic acid encoding a modified fibroin consisting of an amino acid sequence obtained by adding an amino acid sequence comprising an initiation codon and a His10 tag to the N-terminus of the above amino acid sequence is synthesized. May be.
  • the regulatory sequence is a sequence that controls the expression of the modified fibroin in the host (for example, a promoter, an enhancer, a ribosome binding sequence, a transcription termination sequence, and the like), and can be appropriately selected depending on the type of the host.
  • An inducible promoter that functions in a host cell and is capable of inducing the expression of a modified fibroin may be used as the promoter.
  • An inducible promoter is a promoter that can control transcription by the presence of an inducer (expression inducer), the absence of a repressor molecule, or a physical factor such as an increase or decrease in temperature, osmotic pressure, or pH value.
  • the type of expression vector can be appropriately selected depending on the type of host, such as a plasmid vector, a virus vector, a cosmid vector, a fosmid vector, an artificial chromosome vector, and the like.
  • a plasmid vector a virus vector
  • a cosmid vector a fosmid vector
  • an artificial chromosome vector an artificial chromosome vector
  • those capable of autonomous replication in a host cell or integration into a host chromosome and containing a promoter at a position where a nucleic acid encoding a modified fibroin can be transcribed are suitably used.
  • any of prokaryotes and eukaryotes such as yeast, filamentous fungi, insect cells, animal cells, and plant cells can be suitably used.
  • prokaryotic hosts include bacteria belonging to the genus Escherichia, Brevibacillus, Serratia, Bacillus, Microbacterium, Brevibacterium, Corynebacterium and Pseudomonas.
  • microorganisms belonging to the genus Escherichia include, for example, Escherichia coli.
  • microorganisms belonging to the genus Brevibacillus include Brevibacillus agri.
  • Microorganisms belonging to the genus Serratia include, for example, Serratia requestifaciens and the like.
  • microorganisms belonging to the genus Bacillus include, for example, Bacillus subtilis.
  • Microorganisms belonging to the genus Microbacterium include, for example, Microbacterium ammonia phyllum.
  • Examples of microorganisms belonging to the genus Brevibacterium include Brevibacterium divaricatum.
  • Examples of the microorganism belonging to the genus Corynebacterium include Corynebacterium ammoniagenes.
  • Examples of microorganisms belonging to the genus Pseudomonas include Pseudomonas putida.
  • examples of a vector into which a nucleic acid encoding a modified fibroin is introduced include, for example, pBTrp2 (manufactured by Boehringer Mannheim), pGEX (manufactured by Pharmacia), pUC18, pBluescriptII, pSuex, pET22b, pCold, pUB110, pNCO2 (JP-A-2002-238569) and the like.
  • Examples of eukaryotic hosts include yeast and filamentous fungi (such as mold).
  • yeast include yeast belonging to the genus Saccharomyces, the genus Pichia, the genus Schizosaccharomyces, and the like.
  • filamentous fungi include filamentous fungi belonging to the genus Aspergillus, Penicillium, Trichoderma, and the like.
  • examples of a vector into which a nucleic acid encoding a modified fibroin is introduced include YEP13 (ATCC37115) and YEp24 (ATCC37051).
  • any method for introducing the expression vector into the host cell any method can be used as long as it is a method for introducing DNA into the host cell.
  • a method using calcium ions [Proc. ⁇ Natl. ⁇ Acad. ⁇ Sci. USA, 69, 2110 (1972)], electroporation, spheroplast, protoplast, lithium acetate, competent, and the like.
  • a method for expressing a nucleic acid by a host transformed with an expression vector in addition to direct expression, secretory production, fusion protein expression, and the like can be performed according to the method described in Molecular Cloning, 2nd edition, and the like. .
  • the modified fibroin can be produced, for example, by culturing a host transformed with an expression vector in a culture medium, producing and accumulating the modified fibroin in the culture medium, and collecting the modified fibroin from the culture medium.
  • the method of culturing the host in the culture medium can be performed according to a method usually used for culturing the host.
  • the host is a prokaryote such as Escherichia coli or a eukaryote such as yeast, a culture medium containing a carbon source, a nitrogen source, inorganic salts, and the like which can be utilized by the host, so that the host can be cultured efficiently. If so, either a natural medium or a synthetic medium may be used.
  • a prokaryote such as Escherichia coli or a eukaryote such as yeast
  • a culture medium containing a carbon source, a nitrogen source, inorganic salts, and the like which can be utilized by the host, so that the host can be cultured efficiently. If so, either a natural medium or a synthetic medium may be used.
  • the carbon source may be any as long as the transformed microorganism can assimilate, for example, glucose, fructose, sucrose, and molasses containing these, carbohydrates such as starch and starch hydrolyzate, acetic acid and propionic acid Organic acids and alcohols such as ethanol and propanol can be used.
  • the nitrogen source for example, ammonia, ammonium chloride, ammonium sulfate, ammonium salts of inorganic or organic acids such as ammonium acetate and ammonium phosphate, other nitrogen-containing compounds, and peptone, meat extract, yeast extract, corn steep liquor, Casein hydrolyzate, soybean meal, soybean meal hydrolyzate, various fermented cells and digests thereof can be used.
  • potassium (I) phosphate potassium (II) phosphate, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate, and calcium carbonate
  • potassium (I) phosphate potassium (II) phosphate
  • magnesium phosphate magnesium phosphate
  • magnesium sulfate sodium chloride
  • ferrous sulfate manganese sulfate
  • copper sulfate copper sulfate
  • calcium carbonate calcium carbonate
  • ⁇ Cultivation of prokaryotes such as Escherichia coli or eukaryotes such as yeast can be performed under aerobic conditions such as shaking culture or deep aeration stirring culture.
  • the culture temperature is, for example, 15 to 40 ° C.
  • the culturing time is usually 16 hours to 7 days.
  • the pH of the culture medium during the culture is preferably maintained at 3.0 to 9.0.
  • the pH of the culture medium can be adjusted using an inorganic acid, an organic acid, an alkaline solution, urea, calcium carbonate, ammonia, or the like.
  • antibiotics such as ampicillin and tetracycline may be added to the culture medium.
  • an inducer may be added to the medium as necessary.
  • isopropyl- ⁇ -D-thiogalactopyranoside or the like is used.
  • An acid or the like may be added to the medium.
  • the expressed and modified fibroin can be isolated and purified by a commonly used method. For example, when the modified fibroin is expressed in a lysed state in the cells, after completion of the culture, the host cells are collected by centrifugation, suspended in an aqueous buffer, and then sonicated with a sonicator, French press, Menton. The host cells are crushed with a Gaulin homogenizer, Dynomill or the like to obtain a cell-free extract.
  • a method commonly used for isolating and purifying proteins that is, a solvent extraction method, a salting-out method using ammonium sulfate, a desalting method, an organic solvent Precipitation method, anion-exchange chromatography using a resin such as diethylaminoethyl (DEAE) -Sepharose, DIAION @ HPA-75 (manufactured by Mitsubishi Kasei), and cation using a resin such as S-Sepharose @ FF (manufactured by Pharmacia).
  • a resin such as diethylaminoethyl (DEAE) -Sepharose, DIAION @ HPA-75 (manufactured by Mitsubishi Kasei)
  • cation using a resin such as S-Sepharose @ FF (manufactured by Pharmacia).
  • Electrophoretic methods such as ion exchange chromatography, hydrophobic chromatography using resins such as butyl sepharose and phenyl sepharose, gel filtration using molecular sieves, affinity chromatography, chromatofocusing, isoelectric focusing, etc. Purification using methods such as alone or in combination It is possible to obtain the goods.
  • the modified fibroin When the modified fibroin is expressed by forming an insoluble form in the cells, the host cells are similarly recovered, crushed, and centrifuged to collect the insoluble form of the modified fibroin as a precipitate fraction.
  • the recovered insoluble form of the modified fibroin can be solubilized with a protein denaturant.
  • a purified sample of the modified fibroin can be obtained by the same isolation and purification method as described above.
  • the modified fibroin When the modified fibroin is secreted extracellularly, the modified fibroin can be recovered from the culture supernatant. That is, a culture supernatant is obtained by treating the culture by a method such as centrifugation, and a purified sample can be obtained from the culture supernatant by using the same isolation and purification method as described above.
  • the artificial fibroin fibril according to the present embodiment is obtained by spinning the above-mentioned modified fibroin, and contains the above-mentioned modified fibroin as a main component.
  • the artificial fibroin fiber according to the present embodiment is a fiber after spinning and before contact with water.
  • the artificial fibroin fibrils according to the present embodiment can be manufactured by a known spinning method. That is, for example, first, modified fibroin produced according to the above-described method is dissolved in a solvent such as dimethyl sulfoxide (DMSO), N, N-dimethylformamide (DMF), or hexafluoroisopronol (HFIP) as a dissolution promoter. And dissolve it to prepare a dope solution. Next, using this dope solution, spinning is performed by a known spinning method such as wet spinning, dry spinning, dry-wet spinning, or melt spinning to obtain a desired artificial fibroin fibril. Preferred spinning methods include wet spinning and dry-wet spinning.
  • DMSO dimethyl sulfoxide
  • DMF N, N-dimethylformamide
  • HFIP hexafluoroisopronol
  • Preferred spinning methods include wet spinning and dry-wet spinning.
  • FIG. 6 is an explanatory view schematically showing an example of a spinning apparatus for producing artificial fibroin fibrils.
  • the spinning device 10 shown in FIG. 6 is an example of a spinning device for dry-wet spinning, and includes an extruder 1, an undrawn yarn manufacturing device 2, a wet heat drawing device 3, and a drying device 4.
  • a spinning method using the spinning device 10 will be described.
  • the dope solution 6 stored in the storage tank 7 is pushed out of the base 9 by the gear pump 8.
  • a dope solution may be filled in a cylinder and extruded from a nozzle using a syringe pump.
  • the extruded dope solution 6 is supplied into the coagulation solution 11 of the coagulation solution tank 20 via the air gap 19, the solvent is removed, and the modified fibroin is coagulated to form a fibrous coagulate.
  • the fibrous coagulated material is supplied into the hot water 12 in the stretching bath 21 and stretched.
  • the stretching ratio is determined by the speed ratio between the supply nip roller 13 and the take-off nip roller 14.
  • the drawn fibrous coagulated material is supplied to the drying device 4 and dried in the yarn path 22, and the artificial fibroin raw fiber is obtained as the wound yarn 5.
  • 18a to 18g are yarn guides.
  • the coagulating liquid 11 may be any solvent that can remove the solvent, and examples thereof include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol and 2-propanol, and acetone.
  • the coagulating liquid 11 may appropriately contain water.
  • the temperature of the coagulating liquid 11 is preferably 0 to 30 ° C.
  • the extrusion speed is preferably 0.2 to 6.0 ml / hour per hole, and 1.4 to 4.0 ml / hour. More preferably, it is time.
  • the distance that the coagulated protein passes through the coagulating liquid 11 may be long enough to efficiently remove the solvent, for example, 200 to 500 mm. It is.
  • the take-up speed of the undrawn yarn may be, for example, 1 to 20 m / min, and is preferably 1 to 3 m / min.
  • the residence time in the coagulating liquid 11 may be, for example, 0.01 to 3 minutes, and is preferably 0.05 to 0.15 minutes.
  • stretching pre-stretching
  • the coagulation liquid tank 20 may be provided in multiple stages, and the stretching may be performed in each stage or a specific stage as needed.
  • the stretching performed when obtaining the artificial fibroin fibrils includes, for example, dry stretching in addition to wet stretching performed in the coagulation bath 20 and stretching bath 21 described above.
  • Wet heat stretching can be performed in warm water, a solution obtained by adding an organic solvent or the like to warm water, or steam heating.
  • the temperature may be, for example, 50 to 90 ° C, preferably 75 to 85 ° C.
  • the undrawn yarn (or pre-drawn yarn) can be drawn, for example, 1 to 10 times, and preferably 2 to 8 times.
  • Dry heat drawing can be performed using an electric tube furnace, a dry heat plate or the like.
  • the temperature may be, for example, 140 ° C. to 270 ° C., preferably 160 ° C. to 230 ° C.
  • the undrawn yarn (or pre-drawn yarn) can be drawn, for example, 0.5 to 8 times, and preferably 1 to 4 times.
  • the wet heat stretching and the dry heat stretching may each be performed alone, or may be performed in multiple stages or in combination. That is, the first stage stretching is performed by wet heat stretching, the second stage stretching is performed by dry heat stretching, or the first stage stretching is performed by wet heat stretching, the second stage stretching is performed by wet heat stretching, and further the third stage stretching is performed by dry heat stretching.
  • wet heat stretching and dry heat stretching can be performed in an appropriate combination.
  • the final draw ratio is preferably such that the lower limit is more than 1 time, 2 times or more, 3 times or more, 4 times or more, 5 times or more, and 6 times of the undrawn yarn (or pre-drawn yarn). Or more, 7 times or more, 8 times or more, 9 times or more, and the upper limit is preferably 40 times or less, 30 times or less, 20 times or less, 15 times or less, 14 times or less, 13 times or less. , 12 times or less, 11 times or less, 10 times or less.
  • the artificial fibroin fiber is a fiber spun at a draw ratio of 2 or more, the shrinkage when the artificial fibroin fiber is brought into a wet state by contact with water becomes higher.
  • the artificial fibroin fiber according to the present embodiment can be obtained by a manufacturing method including a shrinking step of shrinking the artificial fibroin raw fiber with water.
  • the shrinking step may include, for example, a step (contact step) of bringing the aforementioned artificial fibroin fibrils (fibers after spinning and before contact with water) into contact with water to shrink irreversibly.
  • the shrinking step may include, after the contacting step, a step of drying and further shrinking the fiber (drying step).
  • FIG. 7 is a diagram showing an example of length changes of artificial fibroin fibrils and artificial fibroin fibers due to contact with water.
  • the artificial fibroin fibrils according to the present embodiment have the property of contracting irreversibly (contact length change in FIG. 7) by contact (wetting) with water. After the primary shrinkage, it is further shrunk when dried (the length change indicated by “secondary shrinkage” in FIG. 7).
  • the artificial fibroin fiber obtained through the primary shrinkage or the secondary shrinkage is brought into contact with water and brought into a wet state, it expands to the same length as that before the secondary shrinkage or to a length similar to that before the secondary shrinkage.
  • the contraction and the elongation are repeated with a width approximately equal to the secondary contraction (the width indicated by “stretch ratio” in FIG. 7).
  • Irreversible shrinkage of the artificial fibroin fibrils in the contact step (“primary shrinkage” in FIG. 7) is considered to occur, for example, for the following reasons. That is, one reason is considered to be due to the secondary structure or tertiary structure of artificial fibroin fibrils, and another reason is, for example, in artificial fibroin fibrils having residual stress due to stretching in the manufacturing process. It is considered that water is infiltrated into the fibers or into the fibers, so that the residual stress is reduced. Therefore, it is considered that the shrinkage rate of the artificial fibroin fibrils in the shrinking step can be arbitrarily controlled in accordance with, for example, the magnitude of the draw ratio in the process of producing the artificial fibroin fibrils described above.
  • the artificial fibroin fibrils before spinning are brought into contact with water after spinning to bring the artificial fibroin fibrils into a wet state.
  • the wet state means a state where at least a part of the artificial fibroin fibrils is wet with water. Thereby, the artificial fibroin fibrils can be shrunk without external force. This contraction is irreversible (corresponding to "primary contraction" in FIG. 7).
  • the temperature of the water contacted with the artificial fibroin fibrils in the contacting step may be lower than the boiling point. Thereby, the handleability, the workability in the shrinking step, etc. are improved.
  • the lower limit of the water temperature is preferably 10 ° C. or higher, more preferably 40 ° C. or higher, and further preferably 70 ° C. or higher.
  • the upper limit of the water temperature is preferably 90 ° C. or less.
  • a method of bringing water into contact with artificial fibroin fibrils is not particularly limited.
  • the method for example, a method of immersing artificial fibroin fibrils in water, a method of spraying water on artificial fibroin fibrils at room temperature or in a state of heated steam, and a method of filling artificial fibroin fibrils with water vapor Exposure to a high humidity environment.
  • the method of immersing the artificial fibroin fibrils in water is preferable because the contraction time can be effectively shortened and the processing equipment can be simplified.
  • the artificial fibroin fibrils when the artificial fibroin fibrils are brought into contact with water in a relaxed state, the artificial fibroin fibrils may not only shrink but also shrink in a wavy manner.
  • the artificial fibroin fibrous fibers are brought into contact with water while being tensioned (pulled) in the fiber axis direction, and the contact step is performed in a state where the artificial fibroin fibrils are not relaxed. Is also good.
  • the method for producing artificial fibroin fibers according to the present embodiment may further include a drying step.
  • the drying step is a step of drying and further shrinking the artificial fibroin fibrils that have passed through the contacting step (or the artificial fibroin fibers obtained through the contacting step) (corresponding to “secondary shrinkage” in FIG. 7). Drying may be, for example, natural drying or forced drying using a drying facility. As the drying equipment, any known contact-type or non-contact-type drying equipment can be used. Also, the drying temperature is not limited at all, for example, as long as the protein contained in the artificial fibroin fibrils is decomposed or a temperature lower than the temperature at which the artificial fibroin fibrils are thermally damaged.
  • the temperature is in the range of 20 to 150 ° C, and preferably in the range of 50 to 100 ° C. Temperatures in this range allow the fibers to dry more quickly and efficiently without thermal damage to the fibers or degradation of the proteins contained in the fibers.
  • the drying time is appropriately set according to the drying temperature and the like, and, for example, a time that can minimize the influence of the overdrying on the quality and physical properties of the artificial fibroin fiber and the like is adopted.
  • FIG. 8 is an explanatory view schematically showing an example of a production apparatus for producing artificial fibroin fibers.
  • the manufacturing apparatus 40 shown in FIG. 8 includes a feed roller 42 for feeding out artificial fibroin fibers, a winder 44 for winding the artificial fibroin fibers 38, a water bath 46 for performing a contacting step, and a dryer 48 for performing a drying step. , Is configured.
  • the feed roller 42 is configured to be capable of mounting a wound of the artificial fibroin fibril 36, and is rotated from an wound of the artificial fibroin fibril 36 by rotation of an electric motor (not shown). 36 can be continuously and automatically sent out. After being fed from the feed roller 42, the winder 44 can continuously and automatically wind the artificial fibroin fiber 38 manufactured through the contacting step and the drying step by rotation of an electric motor (not shown). .
  • the feeding speed of the artificial fibroin fiber 36 by the feed roller 42 and the winding speed of the artificial fibroin fiber 38 by the winder 44 can be controlled independently of each other.
  • the water bath 46 and the dryer 48 are arranged between the feed roller 42 and the winder 44 on the upstream side and the downstream side in the feed direction of the artificial fibroin fibril 36, respectively.
  • the manufacturing apparatus 40 shown in FIG. 8 has relay rollers 50 and 52 for relaying the artificial fibroin raw fibers 36 before and after the contact step running from the feed roller 42 toward the winder 44.
  • the water bath 46 has a heater 54, and the water 47 heated by the heater 54 is stored in the water bath 46.
  • a tension roller 56 is installed in a state of being immersed in water 47.
  • the artificial fibroin fibrils 36 sent out from the feed rollers 42 run toward the winder 44 while being immersed in the water 47 while being wound around the tension rollers 56 in the water bath 46. It has become.
  • the immersion time of the artificial fibroin fibrils 36 in the water 47 is appropriately controlled according to the running speed of the artificial fibroin fibrils 36.
  • the dryer 48 has a pair of hot rollers 58.
  • the pair of hot rollers 58 can be wound around the artificial fibroin fibrils 36 that are separated from the water bath 46 and run toward the winder 44.
  • the artificial fibroin fibrils 36 immersed in water 47 in the water bath 46 are heated by the pair of hot rollers 58 in the dryer 48, dried, and further sent out to the winder 44. It has become.
  • the artificial fibroin fibrils 36 are continuously sent out from the feed roller 42 and immersed in water 47 in a water bath 46.
  • the winding speed of the winder 44 is set lower than the feeding speed of the feed roller 42. Accordingly, the artificial fibroin fibrous material 36 contracts by contact with the water 47 in a state where the artificial fibroin fibrous material 36 is tensioned so as not to relax between the feed roller 42 and the winder 44, so that the occurrence of shrinkage can be prevented.
  • the artificial fibroin fibrils 36 contract irreversibly (corresponding to "primary contraction" in FIG. 7).
  • the artificial fibroin fibrils 36 after contact with the water 47 are heated by the pair of hot rollers 58 of the dryer 48.
  • the artificial fibroin fibrils 36 after contact with the water 47 or the artificial fibroin fibers 38 produced through the contact with the water 47
  • can be dried and further contracted (“secondary contraction” in FIG. 7). ").
  • the artificial fibroin fiber 38 can be further contracted and its length can be kept unchanged.
  • the obtained artificial fibroin fiber 38 is wound by a winder 44 to obtain a roll of the artificial fibroin fiber 38.
  • the artificial fibroin fibrils 36 after being brought into contact with the water 47 by using a drying equipment such as a dry heat plate 64 as shown in FIG. It may be dried. Also in this case, by adjusting the relative speed between the feed speed of the feed roller 42 and the winding speed of the winder 44 in the same manner as in the case where a pair of hot rollers 58 is used as the drying equipment, the artificial fibroin fiber is adjusted. 38 can be further contracted or the length can be unchanged.
  • the drying means is constituted by the dry heat plate 64. Further, the dryer 48 is not essential.
  • the target artificial fibroin fiber 38 can be manufactured automatically, continuously, and extremely easily.
  • FIG. 9 is an explanatory view schematically showing another example of a production apparatus for producing artificial fibroin fibers.
  • FIG. 9A shows a processing apparatus provided in the manufacturing apparatus for performing the contacting step
  • FIG. 9B shows a drying apparatus provided in the manufacturing apparatus for performing the drying step.
  • the manufacturing apparatus shown in FIG. 9 includes a processing apparatus 60 that performs a contacting step on the artificial fibroin fibril 36 and a drying step on the artificial fibroin fibril 36 (or the artificial fibroin fiber 38 manufactured through the contacting step) after the contacting step. And a drying device 62 for making them independent from each other.
  • the processing apparatus 60 shown in FIG. 9A omits the dryer 48 from the manufacturing apparatus 40 shown in FIG. 8, and connects the feed roller 42, the water bath 46, and the winder 44 It has a structure in which the fibroin fibrils 36 are arranged side by side in order from upstream to downstream in the running direction.
  • the artificial fibroin fibrils 36 sent out from the feed roller 42 are immersed in water 47 in a water bath 46 to be contracted. Then, the obtained artificial fibroin fiber 38 is wound up by a winder 44.
  • the drying device 62 shown in FIG. 9B has the feed roller 42 and the winder 44, and a dry heat plate 64.
  • the dry heat plate 64 is disposed between the feed roller 42 and the winder 44 such that the dry heat surface 66 contacts the artificial fibroin fiber 38 and extends in the running direction.
  • the drying device 62 as described above, for example, by controlling the ratio of the feed speed of the feed roller 42 and the winding speed of the winder 44, the artificial fibroin fiber 38 can be further contracted, It can be the same.
  • the artificial fibroin fiber 38 can be dried by the drying device 62 after the artificial fibroin fiber 36 is shrunk by the processing device 60 to obtain the artificial fibroin fiber 38. .
  • the processing device may be constituted only by the water bath 46 by omitting the feed roller 42 and the winder 44 from the processing device 60 shown in FIG.
  • a manufacturing apparatus having such a processing apparatus for example, artificial fibroin fibers are manufactured by a so-called batch method.
  • the drying device 62 shown in FIG. 9B is not essential.
  • the artificial fibroin fiber according to the present embodiment may be crimped by contact with water.
  • the degree of crimping is not limited, and the number of crimps may be, for example, 10 to 100 pieces / 40 mm or 20 to 40 pieces / 40 mm.
  • the artificial fibroin fiber crimped by contact with water can be obtained, for example, by contacting the artificial fibroin fibril with an aqueous medium.
  • the aqueous medium is a liquid or gas (steam) medium containing water (including water vapor).
  • the aqueous medium may be water, or a mixture of water and a hydrophilic solvent.
  • the hydrophilic solvent for example, a volatile solvent such as ethanol and methanol or a vapor thereof can be used.
  • the aqueous medium is preferably a mixed solution of water and ethanol.
  • the artificial fibroin fiber dries quickly and has a softer finish.
  • the ratio of water to the volatile solvent or its vapor is not particularly limited.
  • the mass ratio of water: volatile solvent or its vapor may be 10:90 to 90:10.
  • a known oil agent such as an oil for passage through a process (for example, for antistatic) or a finish may be dispersed in the aqueous medium. That is, instead of the aqueous medium, an oil dispersion containing an aqueous medium and an oil dispersed in the aqueous medium can be used. By using such an oil dispersion, the artificial fibroin fibrils can be crimped, and the oil agent can be adhered to the artificial fibroin fibers obtained by crimping the artificial fibroin fibrils. By attaching the oil agent to the artificial fibroin fiber, various properties can be imparted to the artificial fibroin fiber.
  • the amount of the oil agent is not particularly limited, and may be, for example, 1 to 10% by mass or 2 to 5% by mass based on the total amount of the aqueous medium and the oil agent.
  • the temperature of the aqueous medium may be 10 ° C or higher, 25 ° C or higher, 40 ° C or higher, 60 ° C or higher, or 100 ° C or higher, and may be 230 ° C or lower, 120 ° C or lower, or 100 ° C or lower. More specifically, when the aqueous medium is a gas (steam), the temperature of the aqueous medium is preferably 100 to 230 ° C, more preferably 100 to 120 ° C. When the steam of the aqueous medium is 230 ° C. or lower, heat denaturation of the artificial fibroin fiber can be prevented. When the aqueous medium is a liquid, the temperature of the aqueous medium is preferably 10 ° C.
  • the time for contacting the artificial fibroin fibrils with the aqueous medium is not particularly limited, but may be 30 seconds or more, 1 minute or more, or 2 minutes or more, and is preferably 10 minutes or less from the viewpoint of productivity.
  • the contact of the aqueous medium with the artificial fibroin fibrils may be performed under normal pressure or may be performed under reduced pressure (for example, vacuum).
  • a method of contacting the artificial fibroin fibrils with an aqueous medium a method of immersing the artificial fibroin fibrils in water, a method of spraying the artificial fibroin fibrils with an aqueous medium steam, an environment filled with the aqueous medium steam.
  • Examples include a method of exposing artificial fibroin fibrils.
  • the contact of the aqueous medium with the artificial fibroin fibrils can be performed using a general steam setting device.
  • the steam set device include devices such as a product name: FMSA type steam setter (manufactured by Fukushin Kogyo Co., Ltd.) and a product name: EPS-400 (manufactured by Tsujii Dyeing Machinery Co., Ltd.).
  • the method of crimping the artificial fibroin fibrils by the steam of the aqueous medium the artificial fibroin fibrils are accommodated in a predetermined accommodation room, while the steam of the aqueous medium is introduced into the accommodation room, and the temperature in the accommodation room is increased. Is adjusted to the above-mentioned predetermined temperature (for example, 100 ° C. to 230 ° C.), and steam is brought into contact with the artificial fibroin fibrils.
  • the step of crimping the artificial fibroin fibrils by contact with the aqueous medium is preferably performed in a state where no tensile force is applied to the artificial fibroin fibrils (no tension is applied in the fiber axis direction), or a predetermined size. (Strained by a predetermined amount in the fiber axis direction). At that time, the degree of crimp can be controlled by adjusting the tensile force applied to the artificial fibroin fibrils.
  • a method of adjusting the tensile force applied to the artificial fibroin fibrils for example, by suspending various weights on the artificial fibroin fibrils
  • a method of adjusting the load applied to the artificial fibroin fibrils A method in which both ends are fixed in a state where the artificial fibroin fibrils are slackened, and the amount of slack is variously changed, and the artificial fibroin fibrils are wound around a wound body such as a paper tube or a bobbin, and the winding force at that time (Tightening force on paper tube or bobbin) may be appropriately changed.
  • the artificial fibroin fibrils may be dried.
  • the drying method is not particularly limited, and may be natural drying, or the artificial fibroin fibrils may be forcibly dried using a drying facility.
  • the crimping with the aqueous medium and the subsequent drying can be performed continuously.
  • the artificial fibroin fibrils can be dried by being immersed in an aqueous medium while being sent from a bobbin, and then blown with hot air or sent over a hot roller.
  • the drying temperature is not particularly limited, and may be, for example, 20 to 150 ° C., preferably 40 to 120 ° C., and more preferably 60 to 100 ° C.
  • the process of shrinking artificial fibroin fibrils with water which is performed to obtain artificial fibroin fibers, includes processes other than simply reducing the length of artificial fibroin fibrils (not shrinking). And crimping the artificial fibroin fibrils (to shorten the length by crimping).
  • the shrinking step contact step, drying step
  • shrinking reducing the length
  • the artificial fibroin fibrils with water and the step of crimping can be performed simultaneously or stepwise.
  • the artificial fibroin fiber according to the present embodiment may have a wet shrinkage defined by the following formula of 2% or more.
  • Wet shrinkage ⁇ 1 ⁇ (length of artificial fibroin fiber wetted by contact with water / length of artificial fibroin fibril after spinning and before contact with water) ⁇ ⁇ 100 (%)
  • the artificial fibroin fiber according to this embodiment has a wet shrinkage of 2.5% or more, 3% or more, 3.5% or more, 4% or more, 4.5% or more, 5% or more, and 5.5% or more. Or 6% or more.
  • the upper limit of the wet shrinkage is not particularly limited, but is 80% or less, 60% or less, 40% or less, 20% or less, 10% or less, 7% or less, 6% or less, 5% or less, 4% or less, or 3%. It may be:
  • the artificial fibroin fiber according to the present embodiment has a drying shrinkage of 8% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 37% or more, 38% or more. Or 39% or more.
  • the upper limit of the drying shrinkage is not particularly limited, but may be 80% or less, 70% or less, 60% or less, 50% or less, or 40% or less.
  • the artificial fibroin fiber according to the present embodiment may have a limiting oxygen index (LOI) value of 18 or more, 20 or more, 22 or more, or 24 or more, It may be 26 or more, 28 or more, 29 or more, or 30 or more.
  • LOI limiting oxygen index
  • the LOI value is a value measured in accordance with the Fire and Disaster Management Agency, Dangerous Goods Regulations Section, #Fire and Fire Criterion No. 50, May 31, 1995, based on the test method for synthetic resin having a low granularity or a low melting point.
  • the artificial fibroin fiber according to this embodiment may have a maximum heat of moisture absorption determined by the following formula A of more than 0.025 ° C./g.
  • Maximum heat of moisture absorption ⁇ (Maximum value of sample temperature when sample is placed in low humidity environment until sample temperature reaches equilibrium, and then moved to high humidity environment) ⁇ (Sample is sample Sample temperature when placed in a low humidity environment until the temperature reaches equilibrium and then transferred to a high humidity environment) ⁇ (°C) / sample weight (g)
  • the low humidity environment means an environment at a temperature of 20 ° C. and a relative humidity of 40%
  • the high humidity environment means an environment at a temperature of 20 ° C. and a relative humidity of 90%.
  • the artificial fibroin fiber according to the present embodiment may have a maximum heat of moisture absorption by absorption of not less than 0.026 ° C / g, not less than 0.027 ° C / g, and not less than 0.028 ° C / g. May be 0.029 ° C./g or more, may be 0.030 ° C./g or more, may be 0.035 ° C./g or more, and may be 0.040 ° C./g or more. It may be.
  • the upper limit of the maximum heat of moisture absorption is not particularly limited, but is usually 0.060 ° C./g or less.
  • the artificial fibroin fiber according to the present embodiment may have a heat retention index determined according to the following formula B of more than 0.18.
  • Insulation index Insulation rate (%) / Sample weight (g / m 2 )
  • the heat retention rate (%) means a heat retention rate measured by a dry contact method (temperature: 30 ° C., wind speed: 30 cm / sec), and is calculated by (1 ⁇ a / b) ⁇ 100.
  • a shows the amount of heat dissipated through the test piece
  • b shows the amount of heat dissipated without passing through the test piece.
  • the heat retention can be measured by, for example, a Thermolab II tester or the like.
  • the artificial fibroin fiber according to the present embodiment may have a heat retention index of 0.20 or more, may be 0.22 or more, may be 0.24 or more, may be 0.26 or more, It may be 0.28 or more, may be 0.30 or more, and may be 0.32 or more.
  • the upper limit of the heat retention index is not particularly limited, but may be, for example, 0.60 or less, or 0.40 or less.
  • Animal hair fiber in the present specification means animal hair including wool (wool), and fiber derived therefrom.
  • animal hair fibers for example, wool (wool), cashmere (cashmere goat hair), mohair (angora goat hair), angora (angora rabbit hair), alpaca (camelaceae Vicu ⁇ a alpaca hair), vicu ⁇ a ( Camel (Vicuna vicuna hair), Camel (Camelidae camel hair), Lilla (Camelid llama llama hair), and fibers derived therefrom.
  • the composite yarn (blended yarn, mixed fiber yarn, mixed woven yarn, mixed woven yarn, plied yarn, covering yarn, and the like) according to the present embodiment is prepared according to a conventional method except that the above-described artificial fibroin fiber and animal hair fiber are used. Can be manufactured.
  • the composite yarn according to the present embodiment may further contain other fibers other than the artificial fibroin fiber and the animal hair fiber.
  • Other fibers include, for example, synthetic fibers such as polyester and polyamide fibers, and natural fibers such as cotton, hemp, silk, wool, and cashmere. These other fibers are preferably used as long as the effects of the present invention are not impaired.
  • the content ratio of the artificial fibroin fiber and the animal hair fiber in the composite yarn according to the present embodiment is based on the total amount of the artificial fibroin fiber and the animal hair fiber, and the content of the artificial fibroin fiber may be 10% by mass or more, 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more And may be 80% by mass or more.
  • the upper limit of the content of the artificial fibroin fiber may be 90% by mass or less, 80% by mass or less, or 70% by mass or less based on the total amount of the artificial fibroin fiber and the animal hair fiber. , 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, and 20% by mass or less.
  • the content of the artificial fibroin fiber and the animal hair fiber in the composite yarn according to the present embodiment is, based on the total amount of the composite yarn, the total content of the artificial fibroin fiber and the animal hair fiber may be 50% by mass or more, It may be 60% by mass or more, 70% by mass or more, 80% by mass or more, or 90% by mass or more.
  • the upper limit of the total content of the artificial fibroin fiber and the animal hair fiber is not particularly limited, and may be 100% by mass or 95% by mass or less based on the total amount of the composite yarn.
  • the fabric according to the present embodiment contains the above-described artificial fibroin fiber and animal hair fiber.
  • the fabric includes woven fabric, knitted fabric (knitted fabric), lace, felt, nonwoven fabric, and the like.
  • the fabric according to the present embodiment may further contain fibers other than the above-described artificial fibroin fiber and animal hair fiber.
  • Other fibers include, for example, synthetic fibers such as polyester and polyamide fibers, and natural fibers such as cotton, hemp, silk, wool, and cashmere. These other fibers are preferably used as long as the effects of the present invention are not impaired.
  • the fabric according to the present embodiment may contain artificial fibroin fiber and animal hair fiber, for example, by being manufactured by mixing the above-mentioned artificial fibroin fiber and animal hair fiber.
  • the fabric according to the present embodiment also contains, for example, a yarn formed from the above-described artificial fibroin fiber, and a yarn formed from animal hair fiber, which is mixed to produce the artificial fibroin fiber and the animal hair fiber. It may be a thing.
  • the fabric according to the present embodiment may also include, for example, an artificial fibroin fiber and an animal hair fiber by being manufactured using the composite yarn according to the present invention.
  • the yarn formed from the above-mentioned artificial fibroin fiber and the yarn formed from animal hair fiber may be a single yarn or a composite yarn with other fibers. Further, in any of the production methods, other fibers and / or yarns formed from other fibers may be used in combination, if necessary.
  • the fabric (woven fabric, knitted fabric, lace, felt, non-woven fabric, etc.) according to the present embodiment can be manufactured according to a conventional method except that the above-described fibers and / or yarns are used.
  • the content ratio of the artificial fibroin fiber and the animal hair fiber in the fabric according to the present embodiment may be such that the content of the artificial fibroin fiber is 10% by mass or more, based on the total amount of the artificial fibroin fiber and the animal hair fiber. At least 30% by mass, at least 40% by mass, at least 50% by mass, at least 60% by mass, at least 70% by mass. And may be 80% by mass or more.
  • the upper limit of the content of the artificial fibroin fiber may be 90% by mass or less, 80% by mass or less, or 70% by mass or less based on the total amount of the artificial fibroin fiber and the animal hair fiber. , 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, and 20% by mass or less.
  • the content of the artificial fibroin fiber and the animal hair fiber in the fabric according to the present embodiment may be such that the total content of the artificial fibroin fiber and the animal hair fiber is 50% by mass or more, and % Or more, 70% or more, 80% or more, and 90% or more.
  • the upper limit of the total content of the artificial fibroin fiber and the animal hair fiber is not particularly limited, and may be 100% by mass or 95% by mass or less based on the total amount of the fabric.
  • the cloth according to the present embodiment can be used as a material for clothing, medical supplies, sanitary goods, interior goods, bedding, decorations, bags, accessories, miscellaneous goods, vehicle parts, composite goods with resin and the like, for example. it can.
  • Nucleic acids encoding the designed five types of modified fibroins were respectively synthesized.
  • the nucleic acid was added with an NdeI site at the 5 'end and an EcoRI site downstream of the stop codon.
  • These five types of nucleic acids were respectively cloned into a cloning vector (pUC118). Thereafter, the nucleic acid was treated with NdeI and EcoRI with restriction enzymes, cut out, and recombined into a protein expression vector pET-22b (+) to obtain an expression vector.
  • Escherichia coli BLR (DE3) was transformed with the obtained expression vector.
  • the transformed E. coli was cultured in 2 mL of LB medium containing ampicillin for 15 hours.
  • the culture solution was added to 100 mL of a seed culture medium containing ampicillin (Table 4) so that the OD 600 became 0.005.
  • the temperature of the culture was maintained at 30 ° C., and the flask was cultured until the OD 600 reached 5 (about 15 hours) to obtain a seed culture.
  • the seed culture solution was added to a jar fermenter to which 500 ml of a production medium (Table 5 below) had been added so that the OD 600 was 0.05.
  • the temperature of the culture was maintained at 37 ° C., and the culture was performed at a constant pH of 6.9.
  • the concentration of dissolved oxygen in the culture was maintained at 20% of the saturated concentration of dissolved oxygen.
  • a feed solution (455 g / 1 L of glucose, Yeast Extract 120 g / 1 L) was added at a rate of 1 mL / min.
  • the temperature of the culture was maintained at 37 ° C., and the culture was performed at a constant pH of 6.9. Culture was performed for 20 hours while maintaining the dissolved oxygen concentration in the culture solution at 20% of the dissolved oxygen saturation concentration. Thereafter, 1 M isopropyl- ⁇ -thiogalactopyranoside (IPTG) was added to the culture solution to a final concentration of 1 mM to induce the expression of the desired modified fibroin. Twenty hours after the addition of IPTG, the culture was centrifuged to collect the cells.
  • IPTG isopropyl- ⁇ -thiogalactopyranoside
  • SDS-PAGE was performed using the cells prepared from the culture solution before and after the addition of IPTG, and the appearance of a band having a size corresponding to the desired modified fibroin depending on the addition of IPTG revealed the presence of the modified fibroin of interest. Expression was confirmed.
  • the precipitate after washing is suspended in 8 M guanidine buffer (8 M guanidine hydrochloride, 10 mM sodium dihydrogen phosphate, 20 mM NaCl, 1 mM Tris-HCl, pH 7.0) so as to have a concentration of 100 mg / mL, and then suspended at 60 ° C. For 30 minutes with a stirrer to dissolve. After dissolution, dialysis was performed with water using a dialysis tube (cellulose tube 36/32 manufactured by Sanko Junyaku Co., Ltd.). The white aggregated protein obtained after dialysis was recovered by centrifugation. Water was removed from the collected aggregated proteins using a freeze dryer to obtain a freeze-dried powder of modified fibroin.
  • the resulting modified fibroin solution was used as a dope solution (spinning stock solution), and spin-dried and stretched artificial fibroin fibers were manufactured by dry-wet spinning using a spinning device according to the spinning device 10 shown in FIG.
  • the spinning apparatus used in the spinning apparatus 10 shown in FIG. 6 is a second undrawn yarn manufacturing apparatus (between the undrawn yarn manufacturing apparatus 2 (first bath) and the wet heat drawing apparatus 3 (third bath)). (Second bath).
  • the conditions for dry-wet spinning are as follows. Extrusion nozzle diameter: 0.2mm Liquid and temperature in first to third baths: see Table 6 Total draw ratio: see Table 6 Drying temperature: 60 ° C
  • the artificial fibroin raw fibers obtained in Production Examples 1 to 19 are subjected to a contact step of bringing them into contact with water (hereinafter, may be referred to as “primary shrinkage”).
  • a drying step of drying at room temperature is performed (hereinafter, may be referred to as “secondary shrinkage”) to produce artificial fibroin fibers.
  • the length measurement of the artificial fibroin fiber bundle in water was performed with 0.8 g of lead weight attached to the artificial fibroin fiber bundle in order to eliminate the shrinkage of the artificial fibroin fiber bundle.
  • the primary shrinkage (%) of each artificial fibroin fiber was calculated according to the following formula I.
  • L0 indicates the length of the artificial fibroin fibril bundle (here, 30 cm) after spinning and before contact with water
  • Lw indicates the length of the artificial fibroin fiber bundle that has undergone primary shrinkage.
  • the primary shrinkage is synonymous with the wet shrinkage.
  • Primary shrinkage ⁇ 1 ⁇ (Lw / L0) ⁇ ⁇ 100 (%)
  • the prepared spinning stock solution was filtered at 60 ° C. with a metal filter having an aperture of 5 ⁇ m, and then allowed to stand in a 30 mL stainless syringe to remove bubbles, and then coagulated with 100% by mass methanol from a solid nozzle having a needle diameter of 0.2 mm. Discharged into the bath. The discharge temperature was 60 ° C. After coagulation, the obtained fibrils were wound and air-dried to obtain artificial fibroin fibrils.
  • the obtained artificial fibroin fiber was subjected to a contact step of bringing it into contact with water, and then subjected to a drying step of drying at room temperature, thereby producing an artificial fibroin fiber (raw fiber).
  • a knitted fabric was manufactured by circular knitting using a circular knitting machine using the obtained raw material fibers.
  • the knitted fabric had a thickness of 180 denier and 18 gauges. 20 g was cut out from the obtained knitted fabric to obtain a test piece.
  • the flammability test was conducted in accordance with the Fire and Disaster Management Agency, Fire and Disaster Criminal Control Division No. 50, May 31, 1995, test method for synthetic resin having a granular or low melting point. The test was performed under the conditions of a temperature of 22 ° C., a relative humidity of 45%, and an air pressure of 1021 hPa. Table 11 shows the measurement results (oxygen concentration (%), burning rate (%), reduced burning rate (%)).
  • the knitted fabric made of artificial fibroin fibers containing the modified fibroin had a limiting oxygen index (LOI) value of 27.2.
  • LOI limiting oxygen index
  • the LOI value is 26 or more, it is considered to be flame retardant. From this result, it can be seen that the artificial fibroin fiber has excellent flame retardancy. Therefore, a composite yarn or fabric excellent in flame retardancy can be obtained by using a composite yarn or fabric in which artificial fibroin fibers and animal hair fibers are mixed.
  • the prepared spinning dope was filtered at 60 ° C. through a 5 ⁇ m-aperture metal filter, then left standing in a 30 mL stainless syringe and defoamed.
  • the mass% methanol was discharged into a coagulation bath.
  • the discharge temperature was 60 ° C., and the discharge pressure was 0.3 MPa.
  • the obtained yarn was wound at a winding speed of 3.00 m / min and air-dried to obtain artificial fibroin fibrils.
  • the obtained artificial fibroin fiber was subjected to a contact step of bringing it into contact with water, and then subjected to a drying step of drying at room temperature, thereby producing an artificial fibroin fiber (raw fiber).
  • a knitted fabric was manufactured by flat knitting using a flat knitting machine.
  • the knitted fabric using PRT918 fiber as the raw material fiber had a thickness of 1 / 30N (hair count single yarn) and a gauge number of 18.
  • the knitted fabric using PRT799 fiber as the raw material fiber had a thickness of 1 / 30N (hair count single yarn) and a gauge number of 16.
  • the thickness and the number of gauges of the knitted fabric using other raw material fibers were adjusted so as to have almost the same cover factor as that of the knitted fabric using PRT918 fiber and PRT799 fiber. Specifically, it is as shown below.
  • test piece Two pieces of the knitted fabric cut to 10 cm ⁇ 10 cm were aligned, and four sides were sewn to obtain a test piece (sample). After leaving the test specimen in a low humidity environment (temperature 20 ⁇ 2 ° C, relative humidity 40 ⁇ 5%) for 4 hours or more, it was moved to a high humidity environment (temperature 20 ⁇ 2 ° C, relative humidity 90 ⁇ 5%) The temperature was measured at 1 minute intervals for 30 minutes using a temperature sensor attached to the center of the inside.
  • FIG. 10 is a graph showing an example of the result of the moisture absorption / heating test.
  • the horizontal axis of the graph represents the time (minute) of leaving the sample in the high humidity environment as 0 when the time when the sample was transferred from the low humidity environment to the high humidity environment.
  • the vertical axis of the graph indicates the temperature (sample temperature) measured by the temperature sensor.
  • the point indicated by M corresponds to the maximum value of the sample temperature.
  • Table 12 shows the calculation result of the maximum heat of moisture absorption.
  • the artificial fibroin fiber containing the modified fibroin has a higher maximum heat of moisture absorption and is superior in heat absorption and heat generation as compared with existing materials. Therefore, by mixing artificial fibroin fibers and animal hair fibers to form a composite yarn or fabric, a composite yarn or fabric excellent in heat absorption and heat generation can be obtained.
  • the prepared spinning stock solution was filtered at 60 ° C. with a metal filter having an aperture of 5 ⁇ m, and then allowed to stand in a 30 mL stainless syringe to remove bubbles, and then coagulated with 100% by mass methanol from a solid nozzle having a needle diameter of 0.2 mm. Discharged into the bath. The discharge temperature was 60 ° C. After coagulation, the obtained raw yarn was wound up and air-dried to obtain an artificial fibroin fiber (raw material fiber).
  • a knitted fabric was manufactured by flat knitting using a flat knitting machine.
  • the knitted fabric using PRT966 fiber as a raw material fiber had a count of 30 Nm, the number of twists: 1, the number of gauges: 18 GG, and the basis weight: 90.1 g / m 2 .
  • the knitted fabric using PRT799 fiber as the raw material fiber had a count of 30 Nm, a number of twists of 1, a gauge number of GG: 16, and a basis weight of 111.0 g / m 2 .
  • the thickness and the number of gauges of the knitted fabric using other raw material fibers were adjusted so as to have almost the same cover factor as that of the knitted fabric using PRT966 fiber and PRT799 fiber.
  • Insulation index Insulation rate (%) / Sample weight (g / m 2 )
  • Table 13 shows the calculation results of the heat retention index. The higher the heat retention index, the more the material can be evaluated as having excellent heat retention.
  • the artificial fibroin fiber containing the modified fibroin has a higher heat retention index and is superior in heat retention as compared with existing materials. Therefore, by mixing artificial fibroin fiber and animal hair fiber to form a composite yarn or fabric, a composite yarn or fabric excellent in heat retention can be obtained.

Landscapes

  • Engineering & Computer Science (AREA)
  • Textile Engineering (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Mechanical Engineering (AREA)
  • Peptides Or Proteins (AREA)
  • Yarns And Mechanical Finishing Of Yarns Or Ropes (AREA)
  • Woven Fabrics (AREA)
  • Knitting Of Fabric (AREA)
  • Chemical Or Physical Treatment Of Fibers (AREA)
  • Artificial Filaments (AREA)

Abstract

La présente invention concerne un fil composite et un tissu, le fil contenant une fibre de fibroïne artificielle et une fibre de poil animal, la fibre de fibroïne artificielle comprenant une fibroïne modifiée, et ayant un historique de contraction suite à une contraction irréversible en raison du contact avec l'eau après avoir été filée.
PCT/JP2019/038427 2018-09-28 2019-09-27 Fil composite, son procédé de production et tissu WO2020067546A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2018185176A JP2022024192A (ja) 2018-09-28 2018-09-28 複合糸、及びその製造方法、並びに布地
JP2018-185176 2018-09-28

Publications (1)

Publication Number Publication Date
WO2020067546A1 true WO2020067546A1 (fr) 2020-04-02

Family

ID=69951930

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2019/038427 WO2020067546A1 (fr) 2018-09-28 2019-09-27 Fil composite, son procédé de production et tissu

Country Status (2)

Country Link
JP (1) JP2022024192A (fr)
WO (1) WO2020067546A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021241683A1 (fr) 2020-05-28 2021-12-02 シンワ株式会社 Non-tissé et procédé de production dudit non-tissé

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024090523A1 (fr) * 2022-10-26 2024-05-02 Ｓｐｉｂｅｒ株式会社 Fil mélangé élastique et son procédé de production

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106222823A (zh) * 2016-07-28 2016-12-14 东台市苏萌针织时装有限公司 一种多元蛋白调温混纺纱及其制备方法
WO2018164234A1 (fr) * 2017-03-10 2018-09-13 カジナイロン株式会社 Procédé de production de fibre de protéine et procédé de rétraction de fibre de protéine
WO2018164021A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique à retrait élevé ainsi que procédé de fabrication de celles-ci, et procédé de rétraction de fibres de fibroïne synthétique
WO2018165595A1 (fr) * 2017-03-10 2018-09-13 Bolt Threads, Inc. Textiles à base de fibres à supercontraction
WO2018164190A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106222823A (zh) * 2016-07-28 2016-12-14 东台市苏萌针织时装有限公司 一种多元蛋白调温混纺纱及其制备方法
WO2018164234A1 (fr) * 2017-03-10 2018-09-13 カジナイロン株式会社 Procédé de production de fibre de protéine et procédé de rétraction de fibre de protéine
WO2018164021A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique à retrait élevé ainsi que procédé de fabrication de celles-ci, et procédé de rétraction de fibres de fibroïne synthétique
WO2018165595A1 (fr) * 2017-03-10 2018-09-13 Bolt Threads, Inc. Textiles à base de fibres à supercontraction
WO2018164190A1 (fr) * 2017-03-10 2018-09-13 Ｓｐｉｂｅｒ株式会社 Fibres de fibroïne synthétique

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021241683A1 (fr) 2020-05-28 2021-12-02 シンワ株式会社 Non-tissé et procédé de production dudit non-tissé

Also Published As

Publication number Publication date
JP2022024192A (ja) 2022-02-09

Similar Documents

Publication Publication Date Title
US11286286B2 (en) Method and device for manufacturing protein fiber
JP7330468B2 (ja) 混紡糸並びにその編織体及びその編織体の製造方法
JP2020097796A (ja) 人造フィブロイン繊維
WO2020022395A1 (fr) Fibre de cheveux synthétiques, procédé de fabrication associé, et cheveux synthétiques
JP7340262B2 (ja) 高収縮人造フィブロイン紡績糸及びその製造方法、並びに人造フィブロイン紡績糸及びその収縮方法
WO2019194224A1 (fr) Procédé de récupération des dimensions d'un corps de déformation plastique d'un corps moulé en fibroïne modifiée
WO2020067546A1 (fr) Fil composite, son procédé de production et tissu
US20220010460A1 (en) Recombinant-structure protein multifilament and method for manufacturing same
JP7237314B2 (ja) タンパク質繊維の製造方法、タンパク質繊維の製造装置、およびタンパク質繊維の加工方法
WO2020067545A1 (fr) Fil composite et son procédé de fabrication, et tissu
JP7223984B2 (ja) タンパク質紡績糸の製造方法
WO2020067513A1 (fr) Coton en fibre synthétique
WO2020067547A1 (fr) Fibres de fibroïne modifiée
WO2019194263A1 (fr) Fil retors simple de fibroïne synthétique à retrait élevé ainsi que procédé de fabrication de celui-ci, et fil retors simple de fibroïne synthétique ainsi que procédé de rétraction de celui-ci
US20230039765A1 (en) Method for Manufacturing Protein Fiber, Method for Manufacturing Protein Fiber Fabric, and Shrink-Proofing Method for Protein Fiber
WO2019194261A1 (fr) Fibres de fibroïne artificielles
WO2020067553A1 (fr) Procédé de fabrication de fil filé de protéine
JP7367977B2 (ja) タンパク質捲縮ステープルの製造方法
WO2019194230A1 (fr) Étoffe non tissée à haute densité, et procédé de fabrication d'une étoffe non tissée à haute densité
JP7401062B2 (ja) 生地の製造方法
WO2019194260A1 (fr) Fibres artificielles de fibroïne à retrait élevé, leur procédé de production et procédé de rétraction de fibres artificielles de fibroïne
JP2021031811A (ja) 染色された生地の製造方法及び生地の染色方法、並びに染色された混紡糸の製造方法及び混紡糸の染色方法
JP2021054819A (ja) 人工構造タンパク質繊維及びその製造方法
JP2020122251A (ja) 賦形性付与材、賦形性繊維製品及びその製造方法、並びに、形状が付与された繊維製品及びその製造方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19864201

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19864201

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP