WO2022256274A1 - Procédés et systèmes de production de succédané de miel - Google Patents

Procédés et systèmes de production de succédané de miel Download PDF

Info

Publication number
WO2022256274A1
WO2022256274A1 PCT/US2022/031486 US2022031486W WO2022256274A1 WO 2022256274 A1 WO2022256274 A1 WO 2022256274A1 US 2022031486 W US2022031486 W US 2022031486W WO 2022256274 A1 WO2022256274 A1 WO 2022256274A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
operable
vessel
seq
trisaccharides
Prior art date
Application number
PCT/US2022/031486
Other languages
English (en)
Inventor
Efrat Dvash RIESENFELD
Ofir DVASH
Original Assignee
Bee-Io-Honey Technologies Ltd.
The IP Law Firm of Guy Levi, LLC
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 Bee-Io-Honey Technologies Ltd., The IP Law Firm of Guy Levi, LLC filed Critical Bee-Io-Honey Technologies Ltd.
Publication of WO2022256274A1 publication Critical patent/WO2022256274A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
    • A23L21/20—Products from apiculture, e.g. royal jelly or pollen; Substitutes therefor
    • A23L21/25—Honey; Honey substitutes
    • A23L21/27—Honey substitutes

Definitions

  • the disclosure is directed to methods, systems and compositions for producing emulated honey. More specifically, the disclosure is directed to methods systems, and compositions for continuously, batch-wise and semi-continuously producing emulated honey using bioreactors and/or fermenters .
  • honey has been used in medicine since ancient ages and its healing effects on wounds in humans was documented already by the Egyptians at 2000 B.C.
  • honey's therapeutic properties besides osmolarity and acidity are explained by the hydrogen peroxide content as an action of peroxidase oxidase (White, et al. 1963 Biochem Biophys Acta 73, 57-70), the origin of the nectar by its different flavonoid and phenolic acids content (Taormina, et al. 2001. Int J Food Microbiol 69(3), 217-225; Wandan, H. A. 1998. Infection 26(1), 26-31), and an unidentified component (Molan, P. C. 2001.
  • Beekeeping also includes additional abuse such as the removal of the queen-bee wings when it is transferred to a new hive.
  • Honey-bees are often raised at severe conditions including high density, injuries, transportation/handling stress and additional beekeepers’ actions that are applied due to economic considerations only.
  • beekeeping reduces the diversity of wild pollinators and interaction links in the pollination networks. That biodiversity reduction disrupts their hierarchical structural organization causing the loss of interactions by generalist species (e.g., moths, bats), and also impairs pollination services by wild pollinators through reducing the reproductive success of those plant species highly visited by honeybees. High-density beekeeping in natural areas appears to have lasting, serious negative impacts on biodiversity.
  • each vessel is operable to maintain a composition comprising at least one of: a yeast, a fungi, and a bacterium, each transfected with a DNA construct operable to cause the at least one of: the yeast, the fungi, and the bacterium, each operable to express secreted, or cell surface-embedded - at least one predetermined enzyme, wherein the enzyme is at least one of: invertase, amylase, glucose oxidase, superoxide dismutase, catalase, lysozyme, and protease.
  • FIG. 1 is a first implementation schematic of the bioreactor process for producing emulated honey
  • FIG. 2 is a second exemplary implementation schematic of the bioreactor process for producing emulated honey
  • the term “emulated honey” means plant nectar, or its’ equivalent (including among others - sugar solutions) that has undergone a biochemical process resulting in a composition as well as other physico-chemical and organoleptic characteristics that is substantially equivalent to that of honey produced naturally by bees.
  • sugar solution used in certain exemplary implementations can further comprise sugar alcohols, for example: adonitol, allitol, altritol, arabinitol, dulcitol, erythritol, glycerol, iditol, inositol, isomalt, lactitol, maltitol, mannitol, perseitol, ribitol, rhamnitol, sorbitol, threitol or xylitol.
  • sugar alcohols for example: adonitol, allitol, altritol, arabinitol, dulcitol, erythritol, glycerol, iditol, inositol, isomalt, lactitol, maltitol, mannitol, perseitol, ribitol, rhamnitol, sorbitol,
  • the sugar solution used in certain exemplary implementations can comprise indigestible sugars (iS), such as for example: difructose anhydride (DFA) III, fructooligosaccharides (FOS), xylooligosaccharides (XOS), mannanoligo saccharides (MOS), galactooligosaccharides (GOS), and the like.
  • iS indigestible sugars
  • DFA difructose anhydride
  • FOS fructooligosaccharides
  • XOS xylooligosaccharides
  • MOS mannanoligo saccharides
  • GOS galactooligosaccharides
  • system 10 comprising: bioreactor 101 having proximal end 1001 and distal end 1002, bioreactor 101 being in liquid communication with liquid source 100 composition; plurality of vessels 201-204, each vessel 201-204 operable to secrete predetermined enzyme at predetermined period; and collection receptacle 400 and; central processing module (CPM) 700, comprising at least one processor in communication with bioreactor 101, the plurality of vessels 201-204, and collection receptacle 400; CPM 700 being in further communication with memory storage device having thereon processor-readable medium with set of executable instructions, configured when executed, to cause the at least one processor to carry out the steps of the methods disclosed.
  • CPM 700 central processing module
  • each vessel 201- 204 is operable to maintain a composition comprising at least one of: a yeast, a fungi, and a bacterium, each yeast and/or bacterium transfected with a DNA construct operable to cause yeast and/or the bacterium, each operable to express secreted, or cell surface-embedded - at least one predetermined enzyme.
  • the various processes are monitored in several circumstances by a plurality of sensors (see e.g., 150, 450, FIG. 1).
  • the method further comprises contacting vessel (e.g., 201) comprising the at least one of: the yeast, the fungi, and the bacterium operable to express at least one of secreted, and cell surface embedded - at least one predetermined enzyme operable to convert the at least portion of the mono-, di-, and trisaccharides to the substrate comprising hexoses (e.g., aldoses and ketoses), with the liquid source 100 composition for a predetermined period, for example, between about 0.5 min., and about 24 hours (or potentially longer), under controlled temperature, for example between 17 °C, and about 58 °C, thereby forming an inverted liquid source (in other words, plant nectar or it’s equivalent that has undergone invertase digestion).
  • vessel e.g., 201
  • the liquid source 100 composition for a predetermined period, for example, between about 0.5 min., and about 24 hours (or potentially longer), under controlled temperature, for example between 17 °C, and about 58 °C,
  • the invertase used is a fungus (A. niger ) invertase having Protein accession No. ADR80690.1with SEQ ID represented by the following sequence: MKLQT AS VLLGS A A A AS PS MQTRAS VIID YN V APPNLS TLPN GS LFET WRPR AH VLPPN GRS VTPACITPIPPRASFHVGFLHDGSGISSATTDDLPTYQDLNQGNQVIVPGGINDPVAVFDGSV IPN GIN GLPTLLYTS V S YLPIHW S IP YTRGS ETQS LAV S S DGGS NFTKLDQGP VIPGPPF A YN V TAFRDPYVFQNPTLESLLHSKNNTWYTVISGGLHEKGPAQFLYRQYDSDFQYWEYLGQW WHEPTN S T W GN GT W AGRW AFNFET GN VFS LDE Y G YNPHGQ
  • the invertase used is a bacterium (B. subtilis ) invertase having Protein accession No. WP_080528888.1with SEQ ID represented by the following sequence:
  • the invertase used is a Apis mellifera invertase having Protein accession No.
  • the method further comprises contacting vessel 202 comprising the at least one of: the yeast, the fungi, and the bacterium, each operable to express secreted, or cell surface-embedded - at least one predetermined enzyme such as, for example oc-amylase (interchangeable with diastase), operable to convert the at least portion of the starch to a composition comprising mono-, di-, and trisaccharides, with either the preliminary liquid source composition for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature, for example between 17 °C, and about 58 °C.
  • oc-amylase interchangeable with diastase
  • the oc-amylase used is a Rhizopus oryzae oc- amylase having Protein accession No. ADL28123 with SEQ ID represented by the following sequence: MKSFLSLLCSVFLLPLVVQSVPVIKRASASDWENRVIYQLLTDRFAKSTDDTNGCSNLSDYC GGTF QGIINHLD YIAGMGFD AIWIS PIPKN VN GG YHG YW AS DFS QINEHF GT ADDLKKLV A AAHAKNMY VMLD VVANHAGTPS S GGD Y S GYTFGQS SEYHRACDIN YNDQN S IEQC WIS G LPDINTEDS AIV S KLN S IVS GW V S D Y GFDGLRIDT VKH VRKDFWD G Y V S A AG VF ATGE VLS GDVSYVSPYQQHVPSLINYPLYYPVYDVFTKSRTMSRLSSGFSDIK
  • the method further comprises contacting vessel 203 comprising the at least one of: the yeast, the fungi, and the bacterium, each adapted (e.g., through gene transfection) to express secreted, and/or cell-surface embedded at least one predetermined enzyme such as Glucose Oxidase, operable to convert at least one hexose, (e.g., glucose and/or mannose) to an organic acid and hydrogen peroxide, whereby vessel (e.g., 203) is contacted with the inverted liquid source composition after the starch digestion as well, for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature, for example between 17 °C, and about
  • glucose oxidase used is a A. niger glucose oxidase having Protein accession No. AAA32695 with SEQ ID represented by the following sequence:
  • the method further comprises contacting vessel (e.g., 204) comprising the at least one of: the yeast, the fungi, and the bacterium, each operable to express secreted, or cell surface- embedded - at least one predetermined enzyme for example, catalase operable to convert the hydrogen peroxide to tbO and O2, with the converted substrate composition obtained for example, following the digestion with the glucose oxidase secreted from vessel 203 into reactor 101 for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature, for example between 17 °C, and about 58 °C, thereby forming a preliminary processed substrate composition.
  • the 02 is removed from reactor 101 through relief valve 1008, operable to remove excess oxygen from system 10.
  • the catalase used is a Apis mellifera catalase having Protein accession No. NP_001171540 with SEQ ID represented by the following sequence: MTEIKRNPS ADQLID YKKNLKPDCPIFLT GS GTPIS KKAS LTV GPN GPILLQD Y VFLDELS HF NRERIPERVVHAKGAGAFGYFEVTHDITKYSKAKVFSSIGKRTPIAVRFSTVGGESGSADTV RDPRGFA VKF YTEEG VWDLV GNNTPIFFIKDPIYFPS FIHT QKRNP VTHLKD ADMFWDFLS L RPES THQ VMFLFS DRGIPD G YRHMN GY GS HTFS LVN AKDEIV Y C KFH YKTDQGIKNLP VD K AGELSASNPDYAIQDLYDAIAKNQYPTWTFYIQVMTPTQAKSFKWNPFDLTKVWPHDEY
  • the catalase used is a Heliobacter pylori catalase having Protein accession No. AFJ81452 with SEQ ID represented by the following sequence:
  • the protease used is a Apis Mellifera protease (Chymotrypsin) having Protein accession No XP_394370.2 with SEQ ID represented by the following sequence: MIALNVLSVLGFCLAVTAYGVPESKIVGGKNALSGQFPYQVSLRKNKSHFCGGSIIDSRTILT A AHC VEGLS NLN GIT V Q AGTN QLN S GG V S Y VPEKV V AHRS FN ALS LVNDIALIRVN QDIS FT NLIQPIKLAS GS KTYEGSDCILS GW GTTKLN GNVPNNLQWIKLKIETEQKCKQAHWRV QS S HICTFTKS GEGACHGDS GGPLVV GDLQV GIVSFGQPC A V
  • the protease used is a Apis mellifera protease (chymotrypsin-like elastase) having Protein accession No XP_ 006558948.1 with SEQ ID represented by the following sequence:
  • the protease used is a Bacillus subtilis protease (elastase) having Protein accession No AFC89901.1 with SEQ ID represented by the following sequence:
  • the various enzymes are immobilized on the membranes used operably coupled to the reactor 101.
  • the immobilized enzyme can be provided, for example as part of alginate beads.
  • Other immobilization techniques such as, for example immobilization on support surfaces, self immobilization, entrapment in porous (open cell) beads and the like are also contemplated.
  • the enzymes used can be transfected into bacteria and yeast, expressed, or overexpressed either within the periplasm or on the membrane itself (such as for example, optionally chemically modified cell-wall invertase) and be adapted to remain active at high temperatures (in other words, adapted for thermostability), for example, temperatures that will inactivate the host yeast or bacteria (e.g., between about 60 °C and about 90 °C), without substantially adversely affecting the enzymatic activity.
  • substantially adversely affecting enzymatic activity means activity of less than 40% normal activity at STP (1 Atm, at 23 °C).
  • Thermal inactivation of the host bacteria and/or yeast and/or fungi can be done, for example in enzyme reactor 800, maintaining liquid communication with bioreactor 600 (see e.g., FIG. 2) or vessels 201-204 (see e.g., FIG. 1).
  • thermostable enzymes can be isolated from hyperthermophilic bacteria, yeast, fungus or other organisms.
  • Example of enzymes isolated from such hyperthermphilic bacteria can be, for example invertase from Thermotoga maritama, or Pyrococcus furiosus, expressed in, for example Pichia pastoris.
  • the thermostable invertase used is a T. maritama invertase having Protein accession No.
  • thermostable catalase can be used to selectably tune the amount of hydrogen peroxide remaining in the emulated honey produced using the systems and methods provided.
  • Catalases typically found in honey is of pollen origin and its presence modulates the amount of hydrogen peroxide remaining in the produced honey, and which has been found to be beneficial in the bacretriostatic properties of honey.
  • the total concentration of catalase depends on the amount of pollen grains in honey and if hypoallergenic honey is desired, the amount of (added) pollen is drastically reduced.
  • thermostable catalase can be useful in immobilized form in contacting the in-process liquid solutions.
  • the thermostable catalase used is a Bacillus pumilus catalase having Protein accession No. AMW80800.1 with SEQ ID represented by the following sequence:
  • thermostable glucose oxidase can be used in the systems and methods disclosed.
  • the partial concentration of each enzyme and its affinity and reaction kinetics with the in-process liquid can be optimized to produce the proper amount of desired hydrogen peroxide in the emulated honey application where the concentration of hydrogen peroxide and/or the enzymes involved in its production are desired.
  • the thermostable glucose oxidase used is a synthetic thermostable construct with an induced replacement point mutation S100A having Protein accession No. JN809250 with SEQ ID represented by the following sequence:
  • SEQ ID. No. 17 was found to be stable at temperatures of up to 50 °C, without substantially reducing effectiveness for up to 1 hour after transfection with over expression in Pichia pastoris, where wild-type enzyme was sourced from Penicillium notatum.
  • non-peroxide antibacterial properties in honey are attributed inter alia, to the presence of lysozyme sourced from the bees and/or from the nectar (liquid source) used to produce the honey.
  • the enzymes secreted and/or being immobilized and/or expressed on the membrane of the bacteria and/or yeast can be a thermostable gene product sourced for example, from Pseudomonas aeruginosa bacteriophage ⁇ ?KMV having lysozyme domain in its C-terminus with SEQ ID represented by the following sequence:
  • the C-terminus of the gene product represented by SEQ ID. No. 18, is a gene product comprising a highly thermostable lysozyme, retaining 26% of its activity after 2 h at 100°C and 21% after autoclaving (121 -132 °C, at 1 Atm. for 15-60 minutes).
  • expression of secreted, and/or cell-surface, membrane embedded enzymes, by the transfected at least one of yeast, bacteria, and fungi is done by selectively adding at least one of a signal peptide, a promoter, and a terminator to the sequences disclosed herein.
  • a signal peptide added to the protein sequnces disclosed can be SUC2, with SEQ ID represented by the following sequence:
  • a signal peptide sequence having between 80% and 99% homology to SEQ ID. No. 19, its mRNA and cDNA; and/or the signal peptide added to the protein sequnces disclosed can be MATa short and full, with SEQ ID represented by the following sequence:
  • a protein sequence having between 80% and 99% homology to SEQ ID. No. 20, its mRNA and cDNA; and/or the signal peptide added to the protein sequnces disclosed can be MATa mutated, with SEQ ID represented by the following sequence:
  • a protein sequence having between 80% and 99% homology to SEQ ID. No. 21, its mRNA and cDNA; and/or the signal peptide added to the protein sequnces disclosed can be oc-amylase signal protein, with SEQ ID represented by the following sequence:
  • a protein sequence having between 80% and 99% homology to SEQ ID. No. 22, its mRNA and cDNA; and/or the signal peptide added to the protein sequnces disclosed can be lysozyme signaling peptide, with SEQ ID represented by the following sequence:
  • the promoter peptide added to the protein sequnces disclosed can be TEF1, with SEQ ID represented by the following sequence:
  • a protein sequence having between 80% and 99% homology to SEQ ID. No. 24, its mRNA and cDNA; and/or the promoter peptide added to the protein sequnces disclosed can be GPD, with SEQ ID represented by the following sequence:
  • a protein sequence having between 80% and 99% homology to SEQ ID. No. 25, its mRNA and cDNA; and/or the promoter peptide added to the protein sequnces disclosed can be a GAP promoter, with SEQ ID represented by the following sequence:
  • the terminator peptide added to the protein sequnces disclosed can be CYC1 terminator, with SEQ ID represented by the following sequence: [00053] ATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTC CCT ATTT ATTTTTTT AT AGTT AT GTT AGT ATT A AG A AC GTT ATTT AT ATTT C A A ATTTTTC TTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGA AGGTTTT GGG AC GCT C G A AG (SEQ ID. No. 27),
  • a protein sequence having between 80% and 99% homology to SEQ ID. No. 27, its mRNA and cDNA; and or the terminator peptide added to the protein sequnces disclosed can be ADH1 terminator, with SEQ ID represented by the following sequence:
  • the methods implemented using systems 10 disclosed are configured in certain exemplary implementations, with plurality of in-line sensors 150, 450 (and others) operable to analyze a plurality of physico-chemical parameters and provide CPM 700 with the parameters in real time.
  • These plurality of physico-chemical parameters can be, for example at least two of: temperature, pressure, refractive index, turbidity, pH, dynamic viscosity, complex viscosity, flow rate, chroma, and hue.
  • the real-time measurement can then be used to control the residence time of each enzymic process and provide simultaneous control.
  • the parameters measured can provide concentration of reactants and products indicating the end of a reaction, and the like.
  • the at least two sensors can be temperature, refractive index and complex viscosity.
  • CPM may control the processor to increase the temperature of reactor 101.
  • system 10 is in further liquid communication with water source 100, and the method further comprises the steps of using at least one of: the dynamic viscosity sensor, and the complex viscosity sensor, analyzing at least one of: the dynamic, and complex viscosity of the liquid (pure liquid source, inverted liquid source, or processed liquid source e.g.,) in-process; and if the at least one of: the dynamic, and complex viscosity of the liquid in-process is above a predetermined threshold (and temperature is above a predetermined threshold), then, using water source 100, diluting the in-process liquid.
  • the dynamic viscosity sensor and the complex viscosity sensor
  • the system further comprises at least one finishing container 300 comprising a composition of agents operable to provide the preliminary-processed substrate composition with an initial physico-chemical characteristic, forming an intermediary-processed substrate composition.
  • finishing container can comprise a composition of agents operable to provide the preliminary processed substrate with an initial physico-chemical characteristic of a plant-specific honey-emulating components, whereby the plant is, for example, a citrus plant, wild flowers, eucalyptus and the like. Additional agents can be added to fortify the preliminary- processed substrate composition with pharmaceutically active components, flavors, bactericidal agents, exozomes and the like.
  • the preliminary-processed substrate composition optionally with the added agents from finishing container 300, further comprises concentrating the intermediary-processed substrate composition, to obtain a processed liquid source having a final physico-chemical characteristic, in other words, the emulated honey.
  • plurality of vessels 201-204 are each simultaneously coupled to bioreactor 101 via conduit 207k comprising membrane 205i having a predetermined molecular weight cutoff membrane (MWCM) operable to allow passage of predetermined molecular weight proteins therethrough; and flow valve 206j in communication with CPM 700.
  • MWCM molecular weight cutoff membrane
  • Each i Lh membrane 205i can have different MWCM, allowing only reaction product and secreted enzymes to pass through the membrane, while keeping the yeast, bacterium, or fungi (end expressed membrane-embedded enzymes) within their corresponding vessel (201-204).
  • pump 116 may be in communication with vessels 201-204 (or additional vessels) and be also in communication with CPM 700.
  • secreted enzymes in each of vessels 201-204 is collected, ourified and isolated, for example, using various methods, for example tagging with His-tag, and purifiying on Ni-affinity column (as well as other methods similar or identical to those disclosed herein).
  • FIG. 2 Yet another exemplary implementation of the systems implementing the methods disclosed is illustrated in FIG. 2.
  • the step of converting at least the portion of the mono-, di-, and trisaccharides to the substrate comprising hexoses (e.g., aldoses and ketoses) see e.g., FIG.
  • first vessel 601 comprising the at least one of: the yeast, the fungi, and the bacterium, each expressing secreted, or membrane-embedded - at least one predetermined enzyme operable to convert the at least portion of the mono-, di-, and trisaccharides to bioreactor 600 containing the substrate comprising the disaccharides, and the trisaccharides ; and contacting coupled first vessel 601 with the liquid source composition for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature, thereby forming an inverted liquid source.
  • vessel 601 is adapted, sized and configured to form a part of bioreactor 600 and can be comprised of a vessel covered in membrane having a predetermined MWCM, which is configured to allow the liquid source reactants to contact the enzyme secreting medium captured within vessel 601 and the products as well, while compartmentalizing the enzymes within each vessel (601-604). Once it is determined (e.g., using sensors 650 incorporated in the system the reaction is finalized, vessel 601 can be removed and vessel 602 can be reversibly coupled.
  • MWCM a vessel covered in membrane having a predetermined MWCM
  • the step of converting 803 at least a portion of the starch to the composition comprising mono-, di-, and trisaccharides is preceded by: reversibly coupling second vessel 602 comprising the at least one of: the yeast, the fungi, and the bacterium secreting at least one predetermined enzyme operable to convert the starch to bioreactor 600; and contacting second vessel 602 with the liquid source composition for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature (these ranges do not include the time and temperature ranges used in certain implementations, to cause the at least one of the yeast, the fungi, and the bacterium each to express secreted, or membrane-embedded enzymes in an amount operable to perform the steps disclosed, which can be different and longer time-wise).
  • the step of converting at least one hexose to the organic acid and hydrogen peroxide is preceded by: removing the first 601 or second 602 vessel from bioreactor 600; reversibly coupling third vessel 603 comprising the at least one of: the yeast, the fungi, and the bacterium secreting at least one predetermined enzyme operable to convert the at least one hexose to an organic acid and hydrogen peroxide to bioreactor 600; and contacting third vessel 603 with the inverted substrate composition for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature, thereby forming a converted substrate composition.
  • step 805 of converting the hydrogen peroxide to H20 and 02 is preceded by: removing third vessel 603 from bioreactor 600; reversibly coupling to bioreactor 600 fourth vessel 604 comprising the at least one of: the yeast, the fungi, and the bacterium secreting at least one predetermined enzyme operable to convert the hydrogen peroxide to H20 and 02; and contacting fourth vessel 604 with the converted substrate composition for a predetermined period, for example, between about 0.5 min., and about 24 hours, under controlled temperature, thereby forming a preliminary processed substrate composition.
  • operable means the system and/or the device (e.g., nutrient dispensing pump) and/or the program, or a certain element, component or step is/are fully functional, sized, adapted and calibrated, comprising elements for, having the proper internal dimension to accommodate, and meets applicable operability requirements to perform a recited function when activated, coupled or implemented, regardless of being powered or not, coupled, implemented, effected, actuated, realized or when an executable program is executed by at least one processor associated with the system, method, and/or the device.
  • the device e.g., nutrient dispensing pump
  • the program e.g., nutrient dispensing pump
  • a certain element, component or step is/are fully functional, sized, adapted and calibrated, comprising elements for, having the proper internal dimension to accommodate, and meets applicable operability requirements to perform a recited function when activated, coupled or implemented, regardless of being powered or not, coupled, implemented, effected, actuated, realized or when an executable
  • a method for producing emulated honey from a liquid source composition comprising: optionally starch, proteins, trisaccharides, minerals, organic acids and plant pollen, not optionally disaccharides the method comprising: potentially converting at least a portion of the starch to a composition comprising mono-, di-, and trisaccharides using a second enzymatic digestion; not potentially converting at least a portion of the disaccharides, and optionally the trisaccharides to a substrate comprising hexoses (e.g., aldoses, ketoses) using a first enzymatic digestion; converting at least one hexose to an organic acid and hydrogen peroxide using a third enzymatic digestion; optionally converting the hydrogen peroxide to H20 and 02, using a fourth enzymatic digestion; and collecting the converted liquid composition, (i) implemented in a system comprising: a bioreactor having a proximal end and
  • the DNA construct further comprise at least one promoter selected from the promoter group represented by SEQ ID. No.s 24-26, and/or (xxi) the DNA construct further comprise at least one terminator selected from the terminator group represented by SEQ ID. No. 27, or SEQ ID. No. 28, and wherein (xxii) the liquid source is a plant nectar or a composition emulating a plant nectar.
  • the disclosure is directed to systems, compositions and methods for continuously, semi-continuously, batch fed or for that matter, continuous batch, producing emulated honey has been described in terms of some implementations, other implementations will be apparent to those of ordinary skill in the art from the disclosure herein.
  • the described implementations have been presented by way of example only, and are not intended to limit the scope of the inventions. Indeed, the novel methods, programs, libraries and systems described herein may be embodied in a variety of other forms without departing from the spirit thereof. Accordingly, other combinations, omissions, substitutions and modifications will be apparent to the skilled artisan in view of the disclosure herein.

Landscapes

  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

L'invention concerne des procédés, des systèmes et des compositions pour produire un succédané de miel. Plus spécifiquement, l'invention concerne des procédés, des systèmes et des compositions pour produire en continu et semi-continu un succédané de miel à l'aide de bioréacteurs.
PCT/US2022/031486 2021-05-30 2022-05-30 Procédés et systèmes de production de succédané de miel WO2022256274A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202163195028P 2021-05-30 2021-05-30
US63/195,028 2021-05-30

Publications (1)

Publication Number Publication Date
WO2022256274A1 true WO2022256274A1 (fr) 2022-12-08

Family

ID=84324445

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2022/031486 WO2022256274A1 (fr) 2021-05-30 2022-05-30 Procédés et systèmes de production de succédané de miel

Country Status (1)

Country Link
WO (1) WO2022256274A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160243174A1 (en) * 2007-05-03 2016-08-25 Tobias Olofsson Novel bacteria isolated from fresh honey or the honey producing tract of honey bees
US20160289710A1 (en) * 2008-04-30 2016-10-06 Xyleco, Inc. Processing biomass
WO2022155021A1 (fr) * 2021-01-04 2022-07-21 Bee-Io Honey Technologies Ltd. Procédés et systèmes de production de miel émulé

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160243174A1 (en) * 2007-05-03 2016-08-25 Tobias Olofsson Novel bacteria isolated from fresh honey or the honey producing tract of honey bees
US20160289710A1 (en) * 2008-04-30 2016-10-06 Xyleco, Inc. Processing biomass
WO2022155021A1 (fr) * 2021-01-04 2022-07-21 Bee-Io Honey Technologies Ltd. Procédés et systèmes de production de miel émulé

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

Similar Documents

Publication Publication Date Title
Yoshida et al. Microbial activation of two serine enzymes and prophenoloxidase in the plasma fraction of hemolymph of the silkworm, Bombyx mori
MARIN et al. A plant vacuolar system: the lutoids from Hevea brasiliensis latex
Bremberger et al. Dynamics of tonoplast proton pumps and other tonoplast proteins of Mesembryanthemum crystallinum L. during the induction of Crassulacean acid metabolism
Baker et al. Purification, partial characterization, and postembryonic levels of amylases from Sitophilus oryzae and Sitophilus granarius
WO2022256274A1 (fr) Procédés et systèmes de production de succédané de miel
Høidal et al. Mannuronan C‐5 epimerases and cellular differentiation of Azotobacter vinelandii
Srisuparbh et al. Isolation and characterization of major royal jelly cDNAs and proteins of the honey bee (Apis cerana)
Lorito et al. Potential of genes and gene products from Trichoderma sp. and Gliocladium sp. for the development of biological pesticides
Kulakovskaya et al. Inorganic polyphosphates in mitochondria
Pick Adaptation of the halotolerant alga Dunaliella to high salinity
DK167226B1 (da) Fremgangsmaade til selektiv reduktion af proteaseaktivitet i proteaseholdige lactasepraeparater
Sun et al. Polymorphisms in a carboxylesterase gene between organophosphate-resistant and-susceptible Aphis gossypii (Homoptera: Aphididae)
Fulda et al. Isolation of salt-induced periplasmic proteins from Synechocystis sp. strain PCC 6803
Michel et al. Immunocytochemical localization of IdiA, a protein expressed under iron or manganese limitation in the mesophilic cyanobacterium Synechococcus PCC 6301 and the thermophilic cyanobacterium Synechococcus elongatus
KR101880057B1 (ko) 초정탄산수를 함유한 탈모 방지용 헤어케어 화장료 조성물
Cao et al. Differential mRNA expression levels and gene sequences of carboxylesterase in both deltamethrin resistant and susceptible strains of the cotton aphid, Aphis gossypii
Kakimoto et al. Calcium-sensitivity of cortical microtubules in the green alga Mougeotia
CN100381560C (zh) 采用gs115酵母菌合成重组人肠三叶因子的工艺
US5630882A (en) Use of polyether ionophore antibiotics to control bacterial growth in sugar production
KR102365888B1 (ko) 음용수 제조방법 및 이를 이용한 양계 사육방법
FR2573091A1 (fr) Nouvelle enzyme bacteriolytique et son procede de preparation
CN109169645B (zh) 一种用于蝗虫微孢子生物制剂的增效剂
Terra et al. Molecular and evolutionary physiology of insect digestion
Gooding et al. The digestive processes of haematophagous insects: III. Inhibition of trypsin by honey and the possible functions of the Oesophageal diverticula of mosquitoes (diptera)
Keon et al. A comparison of the biochemical and physiological properties of a polygalacturonase from two races of Colletotrichum lindemuthianum

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: 22816696

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE