242 results about "Peptide bond" patented technology

This invention provides a tetrahedral antibody comprising a first, second, third, and fourth domain, wherein the first and second domains are Fab or Fc domains; wherein each of the first and second domains comprise a first polypeptide chain comprising a first N-terminus of the domain, and a second polypeptide chain comprising a second N-terminus of the domain; wherein the first N-terminus of the first domain and the first N-terminus of the second domain are joined to each other by a non-peptidyl linkage, which can be a covalent linkage or a non-covalent linkage between first and second dimerizing polypeptides attached to the first N-termini of the first and second domains, respectively; and wherein the third and fourth domains are attached at their respective C-termini to the second N-termini of the first and second domains, respectively, or the N-termini of the first and second dimerizing polypeptides.

The invention discloses a self-assembly enzyme for directional synthesis of uniform low molecular weight dextran, the self-assembly enzyme is named as YG-SpyTag-SpyCatcher-padex, the YG is a dextran sucrase gene (Genbank No.DQ345760), the padex is a dextran enzyme gene (Genbank No.KF999646.1), the SpyTag and the SpyCatcher are two specific oligopeptides, and aspartic acid in the SpyTag and lysine in the SpyCatcher spontaneously form an isopeptide bond. The invention belongs to the technical field of biology, and relates to an enzyme engineering technology. According to the invention, dextran sucrase derived from an escherichia coli genetically engineered bacterium BL21 (DE3)-YG-SpyTag and dextran enzyme derived from a pichia pastoris genetically engineered bacterium X33-SpyCatcher-padex are combined through a SpyTag-SpyCatcher protein covalent coupling system, and the obtained self-assembled enzyme can be used for catalyzing the substrate sucrose to be directly converted into dextran with the molecular weight of 3000 to 4000 Da.

The invention discloses methanogen lyase homologous with a PeiR lyase sequence and application of the methanogen lyase, and aims to solve the problem that PeiR lyase protein of a methanogen cell wall peptide bond can participate in a biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the PeiR lyase protein has potential methane reduction potential. According to the invention, a series of biological information software is utilized to identify protein sequences coded by methanogens virus from rumen microbiome sequencing data, and sequence homology between the proteins and PeiR protein is further analyzed to obtain a series of methanogens lyases. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through the detection of crude enzyme liquid, the effectiveness of the lyase in the aspect of reducing the methane yield is verified in an in-vitro gas production test.

The invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiW lyase and application of the methanogen lyase, and aims to solve the problem that PeiW lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the protein has potential methane reduction potential. According to the method, a series of bioinformatics software is utilized, a protein sequence coded by methanogens virus is identified from a large amount of rumen microorganism virus group sequencing data, the high-dimensional homology of a deep learning model between the protein sequence and PeiW is analyzed, and two methanogens lyases similar to the high-dimensional characteristics obtained by the PeiW model are obtained. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through detection of a crude enzyme solution, the effectiveness of the lyase in the aspect of reducing the yield of methane is verified in an in-vitro gas production test.

The invention discloses methanogen lyase with the same protein family annotation as PeiR lyase and application of the methanogen lyase, and the methanogen lyase is obtained through homologous screening of Pfam protein family database annotation based on known PeiR lyase characteristics aiming at methanogen cell wall peptide bonds. The PeiR lyase protein can participate in the biological process of hydrolyzing archaea cell walls, methanogens are effectively killed, methane production is reduced, and the protein homologous with the protein has potential methane reduction potential. According to the invention, by integrating metagenome sequencing data of rumen microorganisms, screening methanogenic bacterium virus protein by using series bioinformatics software, and combining Pfam functional domain annotation to carry out homologous relationship analysis, a series of lyase is finally identified and is successfully expressed in a prokaryotic expression system, and meanwhile, an in-vitro gas production experiment result also shows that the methanogenic bacterium virus protein can be successfully expressed in a prokaryotic expression system. The crude enzyme liquid can significantly reduce the methane generation amount.

The invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiR lyase and application of the methanogen lyase and aims to solve the problems that PeiR lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, methanogen is effectively killed, methane production is reduced, and protein homologous with the protein has potential methane reduction potential. According to the invention, a series of biological information software is utilized to identify protein sequences coded by methanogen viruses from a large amount of rumen microbiome sequencing data, and high-dimensional homology between the proteins and PeiR proteins based on a deep learning model TM-Vec is further analyzed, so that a series of methanogen lyases are obtained. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through the detection of crude enzyme liquid, the effectiveness of the lyase in the aspect of reducing the methane yield is verified in an in-vitro gas production test.

The present invention relates to a polymer having a main chain, wherein the main chain comprises linear amide bonds and a heteromonocyclic moiety containing a nitrogen atom, wherein the main chain does not contain peptide bonds, or wherein, based on the total number of linear amide bonds in the main chain, the proportion of peptide bonds in the main chain is at most 30%, and wherein, based on the total weight of the polymer, the proportion of the heteromonocyclic moiety containing a nitrogen atom is from 5% by weight to 25% by weight. The present invention also relates to a product or article and the use of the polymer for manufacturing or producing the product or article.

The present invention relates to vaccine compositions, most particularly vaccine compositions wherein the antigenic component is large (e.g. greater than 50 kDa), or is multimeric (i.e. composed of subunits). Such antigenic components are of particular interest as they may represent antigenic components from pathogens against which vaccines are currently not available. The present invention relates to a composition comprising particles displaying an antigenic component, wherein the composition comprises an antigenic component comprising a first peptide tag and a portion comprising a second peptide tag, wherein the antigenic component and the portion are linked by an isopeptide bond between the first and second peptide tags, and wherein the antigenic component is greater than 50 kDa, or alternatively is multimeric.

The invention discloses a bacterial outer membrane vesicle protein drug delivery system derived from attenuated salmonella as well as a preparation method and application of the bacterial outer membrane vesicle protein drug delivery system. According to the bacterial outer membrane vesicle protein drug delivery system, plug-and-play of a protein drug can be realized on the surface of a bacterial outer membrane vesicle. The invention constructs a bacterial outer membrane vesicle protein drug delivery system. OmpA of attenuated salmonella typhimurium VNP20009 is knocked out, recombinant OmpA of SpyTag inserted into an extracellular ring structural domain is supplemented, fusion protein of SpyCatcher and a protein drug is displayed on the surface of the bacterial outer membrane vesicle by means of the property that an isopeptide bond is spontaneously formed between SpyTag and SpyCatcher, the preparation method is simple and easy to operate, various proteins are displayed on the surface of the bacterial outer membrane vesicle, and the bacterial outer membrane vesicle has a good application prospect. And protein drugs are efficiently protected and loaded. And by combining the immunostimulation characteristic of the bacterial outer membrane vesicles, efficient treatment of tumors is realized.

The present invention relates to a compound binding to an endogenous receptor, said compound comprising (i) an oligopeptide comprising a dipeptide with Trp being the C-terminal amino acid of said dipeptide, wherein said Trp is replaced with an α-amino acid Xaa2, whereby the stability in serum or plasma of the peptide bond connecting Xaa2 to the N-terminally adjacent amino acid is increased as compared to the peptide bond connecting Trp to the N-terminally adjacent amino acid in an otherwise identical compound; and (ii) a moiety capable of generating therapeutically effective radiation, said moiety being covalently bound to said oligopeptide.

The invention provides an extracellular matrix protein recombinant protein, a nucleic acid molecule, a recombinant vector, a genetically engineered bacterium, a preparation method and an application. The provided extracellular matrix protein recombinant protein comprises the following protein fragments: human fibronectin, humanized collagen and elastin-like protein. Wherein the amino acid sequence of the human fibronectin is as shown in SEQ ID NO. 1; the amino acid sequence of the humanized collagen is as shown in SEQ ID NO. 2; the elastin-like protein comprises a pentapeptide repetitive sequence, repetitive units of the pentapeptide repetitive sequence comprise-[valine-proline-glycine-Xaa-glycine] n-, Xaa is isoleucine or lysine, and the number n of the repetitive units is 15-30; in the extracellular matrix protein recombinant protein, any two adjacent ones of the human fibronectin, the humanized collagen and the elastin-like protein are independently connected through a peptide bond or a connecting peptide. The extracellular matrix recombinant fusion protein provided by the invention has good thermal stability and can promote cell adhesion.

The invention relates to the technical field of biology, and particularly discloses a protein-loaded exosome as well as a preparation method and application thereof. The method for preparing the protein-loaded exosome comprises the following steps: (1) mixing a cholesterol-PEG-spy tag / probe tag with the exosome for reaction to obtain the spy tag / probe tag-loaded exosome; and (2) carrying out mixed reaction on the exosome loaded with the spy tag / probe tag and the fusion protein of the target loading protein and the spy catcher / probe, and forming an isopeptide bond through the spy tag / probe tag and the spy catcher / probe, so that the target loading protein is loaded on the exosome. The preparation method provided by the invention can avoid false positive, and the product is single and easy for mass production.

The invention provides an amino acid skeleton ionizable lipid as well as a preparation method and application thereof, the amino acid skeleton ionizable lipid is modified by taking amino acid as a core, has more ester groups and peptide bonds, and can be quickly hydrolyzed by enzyme after RNA is effectively released in vivo; the transfection body is provided with four tail structures, the cross sectional area of the lipid tail can be increased, drugs such as RNA are helped to escape from an endosome, and then the transfection effect is enhanced; the charge capable of ionizing the lipid is electrically neutral under physiological conditions, so that the cytotoxicity caused by excessive positive charges is reduced, the stability of the lipid nanoparticles is further improved, the cycle time of the loaded nucleic acid medicine is prolonged, and the pharmacokinetic characteristics are improved. The LNP prepared from the ionizable lipid, auxiliary phospholipid, cholesterol and PEG lipid provided by the invention has more excellent nucleic acid carrier performance, and can effectively deliver nucleic acid drugs such as siRNA, mRNA, pDNA and the like into cells to play a role.

The present invention relates to a cyclic peptide having the specific formula (I): X1 X2 X3 X4 CX6 X7 X8 PX10 X11 X12 X13 CX15 X16 X17 P (SEQ ID NO: 1) (I) wherein X1 to X17 have various meanings, X1 and D-Pro at position 18 are linked by a peptide bond; the cyclic peptide can be used as an antibacterial agent. The invention also relates to specific bicyclic compounds in which the cysteines in position 5 and 14 are cross-linked by a disulfide bond. The invention also relates to pharmaceutical compositions comprising said peptides and to medical uses of said peptides or compositions comprising them.

There is provided peptidic compounds of Formula I, A or B(Rradn6-[linker]-RL-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-ψ-Xaa9-NH2). Xaa1 is D-Phe, Cpa, D-Cpa, Nal, D-Nal, 2-Nal, or D-2-Nal; Xaa2 is Asn, Gln, Hse, Cit or His. Xaa3 is Trp, Bta, Trp(Me), Trp(7-Me), Trp(6-Me), Trp(5-Me), Trp(4-Me), Trp(2-Me), Trp(7-F), Trp(6-F), Trp(5-F), Trp(4-F), Trp(5-OH), or αMe-Trp. Xaa4 is Ala or Ser. Xaa5 is Val, Cpg, or Tle. Xaa6 is Gly, NMe-Gly, or D-Ala. Xaa7 is His or NMe-His. Xaa8 is Leu or Phe. Xaa9-NH2 is a C-terminally amidated amino acid residue selected from Pro, 4-oxa-L-Pro, Me2 Thz, or Thz. ψ represents a peptide bond or reduced peptide bond joining Xaa8 to Xaa9. Rradn6 is 1-5 radiolabeling groups. There is also provided the use of such compounds as imaging agents or therapeutic agents.

There is provided peptidic compounds of Formula I, A or B(Rradn6-[linker]-RL-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-ψ-Xaa9-NH2). Xaa1 is D-Phe, Cpa, D-Cpa, Nal, D-Nal, 2-Nal, or D-2-Nal; Xaa2 is Asn, Gln, Hse, Cit or His. Xaa3 is Trp, Bta, Trp(Me), Trp(7-Me), Trp(6-Me), Trp(5-Me), Trp(4-Me), Trp(2-Me), Trp(7-F), Trp(6-F), Trp(5-F), Trp(4-F), Trp(5-OH), or αMe-Trp. Xaa4 is Ala or Ser. Xaa5 is Val, Cpg, or Tle. Xaa6 is Gly, NMe-Gly, or D-Ala. Xaa7 is His or NMe-His. Xaa8 is Leu or Phe. Xaa9-NH2 is a C-terminally amidated amino acid residue selected from Pro, 4-oxa-L-Pro, Me2Thz, or Thz. ψ represents a peptide bond or reduced peptide bond joining Xaa8 to Xaa9. Rradn6 is 1-5 radiolabeling groups. There is also provided the use of such compounds as imaging agents or therapeutic agents.

A compound comprising a prostate specific membrane antigen (PSMA)-targeting moiety of the following formula (I) or of a salt or a solvate thereof. R1 is —R1aR1b— wherein R1a is absent, —CH2—, —O—, or —S—, and R1b is —CH2— or —CHF—. R2 is —(CH2)3—O—, —(CH2)3—, —(CH2)4—, —CH2—O—(CH2)2—, or —CH2—S—(CH2)2. R3 is ethylene fused to a tricyclic fused ring. L1, L2, L3, L4, L5b L5c, L6 and L7 are linkages (e.g. peptide bonds). Xbr is a branching atom. Rrad is a radiometal chelator. Ralb is an albumin binder. n1, n2, n3, n4, and n5 are integers. When the PSMA-targeting moiety is linked to a radiolabeling group, the compound may be used as an imaging agent or therapeutic agent for PSMA-expressing diseases / conditions.

The invention relates to the technical field of biology, in particular to a mirror image coupled protein nanoparticle conjugate and application thereof. The conjugate comprises: ii) a functional protein labeled by a first peptide tag, and ii) a scaffold protein labeled by a second peptide tag, the first peptide tag and the second peptide tag being composed of D-amino acid residues; the first peptide tag and the second peptide tag form a peptide partner pair, and the first peptide tag and the second peptide tag are coupled and combined through an isopeptide bond. The conjugate provided by the invention is easier to escape from the recognition of protease, can obviously prolong the half-life period in a biological system, is not easy to cause unnecessary immunoreaction, and has good biological stability and safety.

Compositions and related methods and uses are provided comprising sotebercept and variants thereof wherein the variants undergo a peptide bond cleavage between arginine 4 and serine 5 (Arg (4)-Ser (5) of the amino acid sequence (SEQ ID NO: 2) of one or two monomeric peptide chains of sotebercept, calculated based on peptide mass fingerprinting analysis, and wherein the variants have a peptide bond cleavage between arginine 4 and serine 5 (Arg (4)-Ser (5) of one or two monomeric peptide chains of sotebercept. The variant content is less than or equal to about 10%. The invention further provides a pharmaceutical preparation containing the composition and a preparation method of the pharmaceutical preparation. In addition, the invention also provides a detection method of the variant and application of the variant in quality inspection or quality control of a product containing sotebercept.

The invention relates to molecular glue for forming fimbriae related protein based on SpaA isopeptide bonds of Vinci bacteria. The invention also relates to a nanoparticle vaccine composition formed by the molecular glue, and the immunogen is covalently displayed on the surface of the nanoparticle by the molecular glue.

Crosslinking molecules that permit efficient identification of specifically associated proteins and the sites of crosslinking in biological or other samples include two cleavable bonds that are cleavable under the same conditions as peptide bonds in mass spectrometric determinations. Sets of the invention crosslinkers may be provided with isobaric labels containing reporter ions of differing molecular weights to permit relative quantitation of crosslinked protein pairs.

The invention belongs to the technical field of enzyme engineering and gene engineering, and particularly relates to a recombinant immobilized enzyme, an expression gene thereof and application of the recombinant immobilized enzyme in production of macrocyclodextrin. According to the preparation method, SpyTag and SpyCatcher peptides are fused to the N end of isoamylase and the N end of 4-alpha-glucanotransferase respectively, the 4-alpha-glucanotransferase modified by the SpyTag and the isoamylase modified by the SpyCatcher are obtained, and after spontaneous isopeptide bonds are formed between the SpyTag and the SpyCatcher, the self-assembled macrocyclodextrin production enzyme cluster is obtained. The obtained macrocyclodextrin production enzyme cluster forms an enzyme aggregate with a three-dimensional structure under the traditional glutaraldehyde-mediated catalytic action, namely the recombinant immobilized enzyme. The recombinant immobilized enzyme disclosed by the invention can be repeatedly utilized, is simple and convenient to operate, is easy to separate a converted product, and can be applied to industrial production of the macrocyclodextrin.

The present invention pertains to specific bioparticules at the surface of which are expressed multimeric protein. The bioparticles according to the present invention comprise an envelope consisting of a plasma membrane; and at least one type I or II transmembrane fusion protein anchored in said membrane, said fusion protein comprising successively a) a first monomer of a multimeric protein of interest, b) a coiled-coil domain or oligomerization sequence; and c) a domain for anchoring in the plasma membrane, consisting of a transmembrane segment and a cytosolic segment. Fragments a) and b) are exposed at the surface of the bioparticle, and fragment a) is bound to a second monomer of said multimeric protein by means of a bond which is not a peptide bond. The bioparticles according to the present invention can be used in therapy such as immunotherapy. The present invention also pertains to methods for producing such bioparticles.

The invention discloses a sea-buckthorn small molecule peptide and a preparation method thereof, and relates to the field of biological medicine and food processing. According to the method, sea-buckthorn pulp residues and sea-buckthorn seed residues are used as composite raw materials and are mixed and stirred to form homogenate, and then cell wall structures are destroyed through ultrasonic wall breaking treatment to release protein components; then cutting off a peptide bond under the action of compound enzymes (plant protease, neutral protease and pepsase) by adopting an ultrasonic-enzyme synergistic hydrolysis process to generate a small molecule peptide fragment; after removing impurities through centrifugal separation, purifying through an ultrafiltration membrane (the molecular weight cut-off is 1-3kDa) to obtain a small molecule peptide solution; and performing debitterizing treatment by using macroporous adsorption resin, and preparing sea-buckthorn small molecule peptide powder by using a vacuum freeze-drying technology. According to the method, through ultrasonic-enzyme synergistic interaction, multi-stage purification combination and low-temperature keep-alive processes, the yield, molecular weight uniformity and antioxidant activity of small molecular peptides are remarkably improved, and high-valued utilization of sea-buckthorn processing by-products is achieved.

The invention discloses an in-vitro biochemical test method for evaluating the anti-wrinkle effect of cosmetics and raw materials. The in-vitro biochemical test method comprises the following steps: step 1, preparing a working solution; 2, preparing a sample solution; step 3, adding an enzyme working solution; step 4, loading samples in groups; 5, adding a substrate and reacting; step 6, developing treatment; step 7, determining absorbance; 8, calculating, analyzing and processing data; and step 9, result judgment: analyzing and evaluating the anti-wrinkle effect of the sample through a statistical means. According to the method, based on a collagenase inhibition test, when peptide bonds between GLY and GLY in Z-GLY-PRO-GLY-GLY-PRO-ALA-OH are specifically hydrolyzed by collagenase, GLY-PRO-ALA is released, and the activity of the collagenase is indirectly evaluated by measuring the change of a light absorption value at 565 nm after the GLY-PRO-ALA reacts with a color developing agent, so that whether a sample to be detected has an anti-wrinkle effect or not is judged. The method is economical and fast, the test process is controllable, the test system is standardized, and result deviation caused by individual difference can be avoided.

The invention relates to a surface display system based on an isopeptide bond and a construction method and application of the surface display system, two conformational proteins including an anchor protein containing the isopeptide bond and a target protein are designed through a modularization method, and through covalent coupling formed between lysine and asparagine or aspartic acid residues in a molecular adhesive system based on the isopeptide bond, the surface display system based on the isopeptide bond is obtained. And displaying the target protein on the surface of escherichia coli. According to the surface display system based on the isopeptide bond, high-efficiency display of complex enzyme and macromolecular protein on the cell surface is realized, and the influence of direct fusion with anchoring protein on the activity of target protein is avoided. According to the constructed surface display whole-cell catalyst, the tedious enzyme purification process is omitted, the stability of the enzyme is improved, the production cost is lower, and the surface display whole-cell catalyst shows a good application prospect in biological catalysis.

This invention discloses methanogenic lyases with the same protein family annotation as PeiR lyases and their applications. These enzymes are obtained through homology screening based on the known characteristics of PeiR lyases targeting peptide bonds in the methanogenic cell wall, using the Pfam protein family database annotation. PeiR lyase proteins participate in the biological process of hydrolyzing archaea cell walls, effectively killing methanogens and reducing methane production. Proteins homologous to this protein have potential methane-reducing potential. This invention integrates rumen microbial metagenomic sequencing data, uses a series of bioinformatics software to screen methanogenic viral proteins, and combines Pfam functional domain annotation for homology analysis, ultimately identifying a series of lyases. These lyases were successfully expressed in a prokaryotic expression system, and in vitro gas production experiments also showed that the crude enzyme solution significantly reduced methane production.

This invention belongs to the field of photocrosslinking probe technology, specifically relating to a GUSB photocrosslinking probe, its preparation method, and its application. The GUSB photocrosslinking probe has the general chemical formula C0. 13 H 18 N2O6-R, wherein R is any one of the oxalic group elements or peptide bonds. The R group covalently connects the glucuronic acid recognition core and the photosensitive unit in the GUSB photocrosslinking probe, and the R group contains no more than 4 atoms, resulting in a small size and low steric hindrance. The GUSB photocrosslinking probe provided by this invention uses glucuronic acid as the recognition core, combined with a small-volume diazacyclopropane photosensitive unit and a terminal alkyne group, to achieve µM-level in-situ covalent capture of active GUSB in complex systems such as cells or tissues. The probe of this invention significantly improves positioning accuracy, sensitivity, data availability, quantification, and enrichment, while also possessing advantages such as modular synthesis, mild conditions, and strong system compatibility, effectively addressing the technical shortcomings of existing technologies that struggle to accurately obtain GUSB activity in situ under low-dose conditions.

The invention relates to the technical field of enzyme engineering and food processing, and particularly discloses a method for preparing soybean meal peptide through solid enzymolysis of soybean meal with a compound enzyme. According to the method, alkaline protease, neutral protease and trypsin are compounded into a compound enzyme according to the mass ratio of 1: 1: 1, and the yield and quality of the soybean meal peptide are remarkably improved by optimizing solid enzymolysis process conditions, the temperature is 50 DEG C, the pH is 6.5, the time is 7 hours and the material-liquid ratio is 1: 0.5. The hydrolysis degree reaches 24.61%, the peptide yield reaches 14.77%, the material-liquid ratio is reduced to 1: 0.5 compared with a traditional liquid method, near-zero wastewater discharge is achieved, and the environmental protection cost is reduced. Fourier infrared spectroscopy verifies that the peptide bond structure of the product is complete (the amide I band is 1650 cm <-1 >), and the secondary conformation is mainly beta-folding. The invention provides a low-cost, low-energy-consumption and sustainable soybean meal peptide industrial production scheme for environment-friendly strictly-controlled areas and small and medium-sized enterprises, and has the advantages of high resource value and green process.

This invention discloses a methanogenic lyase homologous to PeiR lyase and its applications. The PeiR lyase protein, targeting the peptide bonds of the methanogenic cell wall, participates in the biological process of hydrolyzing the archaea cell wall, effectively killing methanogens and reducing methane production. Proteins homologous to this lyase possess potential methane-reducing potential. This invention utilizes a series of bioinformatics software to identify protein sequences encoded by methanogenic viruses from a large amount of rumen microbial virome sequencing data, and analyzes the structural homology between these proteins and PeiR. Ultimately, this invention successfully obtained a series of methanogenic lyases homologous to PeiR. To achieve the expression of these lyases, this invention designed multiple primers to synthesize the target sequences and successfully expressed them in a prokaryotic expression system. In vitro gas production experiments with the crude enzyme solution showed that these lyases are effective in reducing methane production.