1025 results about "Hydrolase" patented technology

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

The invention relates to the technical field of microorganisms, in particular to a culture medium and a method for culturing Ackermania muciniphila for activating Amuc-1260 by using lambda-carrageenan as a carbon source, according to the culture method, the lambda-carrageenan is used for replacing a traditional carbon source, thallus growth and proliferation can be remarkably promoted, the effect is optimal under the condition of 2.00 g / L, and the culture cost is low. Compared with the conventional carbon source (glucose), the carbon source is improved by 92.91%. Further research finds that the lambda-carrageenan can induce the Akk bacteria to up-regulate the expression of glycoside hydrolase such as Amuc-1260 and enhance the activity of pathways such as glycosaminoglycan degradation, amino acid metabolism and insulin signals, so that the metabolic adaptability and energy utilization efficiency of the bacteria are improved. Meanwhile, the Akk bacteria secrete more functional short-chain fatty acids such as acetic acid, propionic acid and the like under the condition, and have the effects of regulating intestinal health and maintaining metabolic homeostasis. The invention provides a new theoretical basis and technical support for large-scale culture of Akk bacteria and development of a metabiotic product.

The invention relates to the technical field of microbial genetic engineering, in particular to a method for improving cordycepin synthesized by saccharomyces cerevisiae and saccharomyces cerevisiae engineering bacteria. According to the invention, codon-optimized 2 '-carbonyl-3'-deoxyadenosine reductase gene and 3 '-adenosine monophosphate phosphohydrolase gene from cordyceps militaris are expressed by using plasmids, and then the plasmids are transformed into host cells to obtain the saccharomyces cerevisiae engineering bacteria. And inoculating a seed solution obtained by seed culture of the activated saccharomyces cerevisiae engineering bacteria into a fermentation culture medium containing a metabolic effector, and carrying out fermentation culture to synthesize cordycepin. Metabolic effectors (Cu < 2 + >, Fe < 2 + >, Mg < 2 + >, Zn < 2 + >, citric acid, cysteine, aspartic acid, glycine, VB1 and tea polyphenol) are added into a fermentation culture medium strain, and the variety and concentration of the metabolic effectors are optimized, so that the yield and synthesis efficiency of cordycepin are remarkably improved. By using the combination of the strain obtained by the invention and the optimal effector, the yield of cordycepin in a fermentation tank reaches 2.9 g / L.

The invention provides a preparation method of a protein base material for preparing a protein beverage, which comprises the following steps: adding yeast protein or yeast powder into a liquid, and dispersing and hydrating to form a suspension; adding an enzyme into the suspension for enzymolysis; wherein the enzyme comprises a first enzyme, a second enzyme and a third enzyme; the first enzyme and the second enzyme are protease, and the third enzyme is cellulose hydrolase. According to the method, the solubility and flavor of the yeast protein can be effectively improved, and the yeast protein beverage with excellent flavor and stability is finally obtained. The yeast protein beverage base material which is good in flavor, high in solubility, less in additive and easier to digest and absorb can be prepared through physical and enzymolysis methods which are easy to realize in the food industry without adding other substances.

The invention belongs to the field of biological catalysis, and discloses a high-activity PET hydrolase mutant and application thereof. The mutant of the thermophilic PET hydrolase provided by the invention is as shown in 1), 2) or 3): 1) a protein corresponding to an amino acid sequence SEQ ID NO.4; 2) a protein corresponding to the amino acid sequence of SEQ ID NO.6; and 3) a protein corresponding to the amino acid sequence of SEQ ID NO.8. Experiments prove that the modified mutant shows higher catalytic efficiency on PET hydrolysis and can keep activity for a long time at the optimal temperature for PET depolymerization. The activity of the mutant protein BhrM3 for catalyzing PET hydrolysis is improved by 1.82 times compared with that of a wild type and is 1.22 times that of efficient PET hydrolase LCC-ICCG, and the protein melting temperature reaches 92.9 DEG C.

The invention discloses a group of amide hydrolase mutants capable of efficiently degrading ochratoxin A as well as a coding gene, a recombinant vector and application of the amide hydrolase mutants, and belongs to the technical field of enzyme engineering. The invention also provides application of the mutant of the group of recombinant amide hydrolase in detoxification of ochratoxin A in beer or corn flour polluted by ochratoxin A. The activity of the mutant on ochratoxin A is obviously improved compared with the activity of the existing recombinant amide hydrolase ADH3; beer or corn flour containing ochratoxin A is treated for 10-15 minutes, the degradation rate of ochratoxin A can reach 100%, and the degradation efficiency is high in the field.

The invention discloses a methanophilic genetically engineered bacterium for producing trehalose as well as a construction method and application of the methanophilic genetically engineered bacterium, and belongs to the technical field of industrial biology. The methanophilic genetically engineered bacterium capable of producing the trehalose is obtained by taking methanophilic bacteria methanophilic 5GB1, Alkalicicus glycogenes WONF2802 or methanophilic bacteria NG09 as host bacteria, introducing maltooligosyl trehalose synthase (MTS) into the host bacteria, and carrying out over-expression on maltooligosyl trehalose hydrolase (MTH), so as to obtain the methanophilic genetically engineered bacterium capable of producing the trehalose. Under the condition that methane or methanol serves as a carbon source, the yield of the produced trehalose can reach the gram level or above, a new low-carbon biosynthesis way can be provided for commercial production of the trehalose, and the methanophilic genetically engineered bacterium for producing the trehalose has a good industrial application prospect.

The invention discloses an ochratoxin A degrading enzyme mutant and application thereof. The invention provides mutant protein of amidase MiADH, which is any one of the following changes: the amino acid sequence of the amidase MiADH as shown in SEQ ID NO: 1 is subjected to any one of the following changes: a 66th amino acid residue Y is mutated into K, or a 143rd amino acid residue H is mutated into A, or a 194th amino acid residue V is mutated into A, or a 195th amino acid residue L is mutated into A, or a 230th amino acid residue H is mutated into A, or the amino acid sequence of the amidase MiADH is mutated into A, or the amino acid sequence of the amidase MiADH is mutated into K, or the amino acid sequence of the amidase MiADH is mutated into A; or the 324th amino acid residue V is mutated into A, and the protein has the same function; experiments prove that the amide hydrolase MiADH mutant can efficiently degrade OTA, and OTA detoxification can be achieved under the high-temperature condition.

The invention discloses a PET hydrolase mutant as well as a preparation method and application thereof, and belongs to the field of enzyme engineering. On the basis of FAST-PETase, in combination with deep learning prediction and enzyme engineering optimization strategies, a plurality of mutants with specific performance advantages are systematically constructed and screened, the method is suitable for application scenes with different temperatures, substrate loads and degradation efficiency requirements, and experimental basis and technical support are provided for constructing a diversified and combinable PET degrading enzyme system. The mutants show diversified advantages in different performance dimensions, for example, the enzyme activity of the K233R mutant is 1.62 times that of FAST-PETase, and the K233R mutant still keeps good thermal stability (1.03 times) after being subjected to heat treatment for 2 hours at the temperature of 50 DEG C; q224K is more remarkably improved in the aspect of thermal stability, and the residual activity after heat treatment is 1.64 times that of FAST-PETase.

The invention relates to a recombinant pichia pastoris genetically engineered bacterium for expressing recombinant full-chain-length humanized collagen as well as a preparation method and application of the recombinant pichia pastoris genetically engineered bacterium, and belongs to the technical field of genetic engineering. The recombinant pichia pastoris genetically engineered bacterium disclosed by the invention expresses recombinant full-chain-length humanized collagen, and a ku70 gene of the recombinant pichia pastoris genetically engineered bacterium is knocked out and knocked into a recombinant 4-proline hydroxylase encoding gene. Recombinant full-chain-length humanized collagen is optimized, a 4-proline hydroxylase gene and a collagen gene are co-expressed in pichia pastoris to obtain hydroxylation modified recombinant collagen, and a corresponding recombinant pichia pastoris genetically engineered bacterium is provided, has proteolytic enzyme A defect and hydroxylation modification capability, and can be used for preparing a recombinant collagen product. The stability of the collagen in the fermentation process is improved. The recombinant full-chain-length humanized collagen provided by the invention is 100% homologous with III-type collagen derived from a human body, so that the immune risk is extremely low, and the biological activity is more perfect.

This invention relates to the field of water treatment technology, specifically to a reverse osmosis membrane cleaning agent composed of a non-oxidizing bactericide and an enzyme, and its application. Its main purpose is to remove biofouling from reverse osmosis membranes and restore membrane flux. This patent discloses a novel reverse osmosis membrane cleaning agent formulation. Considering the different pH requirements of the cleaning agent in actual production, different types of bioactive hydrolytic enzymes are selected to disrupt the structure of bacterial cell walls and their extracellular polymers. A reducing agent and a non-oxidizing bactericide are combined to enhance the cleaning effect of the reverse osmosis membrane. For different cleaning purposes, non-oxidizing bactericides suitable for acidic and alkaline cleaning are formulated. Sodium benzoate and benzalkonium chloride show good application prospects in the removal of reverse osmosis membrane fouling.

The invention relates to a method for detecting activity of gamma-polyglutamic acid hydrolase and application. The method comprises the following steps: (1) preparing an enzymolysis reaction solution and a blank reaction solution; (2) preparing an NAC / OPA mixed solution; (3) adding an NAC / OPA mixed solution into the enzymatic hydrolysis reaction solution to obtain a reaction solution to be detected; (4) drawing a standard curve by taking L-glutamic acid as a standard substance; and (5) detecting the fluorescence intensity of the blank reaction solution and the reaction solution to be detected by using a microplate reader, and then calculating the activity of the gamma-polyglutamic acid hydrolase according to the fluorescence intensity and the standard curve. The method for detecting the activity of the gamma-polyglutamic acid hydrolase is high in detection speed, detection of a single sample can be completed within 25 min, and nearly real-time process monitoring is achieved. And the method has the advantages of reliable detection result and high accuracy, can be used for monitoring the enzyme activity of the gamma-polyglutamic acid hydrolase in the gamma-polyglutamic acid preparation process in real time and guiding the optimization of the fermentation process, and has extremely high industrial application value.

The invention discloses a Vinck Cellulomicrobe and application thereof in degradation of corn straw and improvement of soil fertility, and belongs to the field of agricultural microbial technology and solid waste resource utilization. The bacterial strain can secrete a plurality of hydrolase and oxidase including AA10 family lytic polysaccharide monooxygenase, AA2 peroxidase, GH3 and GH1 beta-glucosidase, GH16 / GH17 beta-glucosan endonuclease and AA1 multi-copper oxidase. The bacterial strain has the advantages that the bacterial strain can secrete a plurality of hydrolase and oxidase; fermentation liquor is applied to soil according to the straw mass ratio of 1: 5, and the corn straw degradation rate can be increased under the condition that the mass ratio of soil to straw is 100: 1. Meanwhile, the microbial agent can synergistically improve soil nutrients and increase the contents of rapidly available phosphorus, rapidly available potassium, alkali-hydrolyzable nitrogen, soluble organic carbon and total phosphorus in soil. According to the microbial agent, a straw lignocellulose structure is effectively cracked through a multi-enzyme system synergistic effect, and the dual effects of accelerating straw degradation and improving soil fertility are achieved.

The invention discloses a method for producing D-pantoic acid by immobilizing D-pantoic acid lactone hydrolase fusion protein, and belongs to the technical field of biology. The insoluble seed pumpkin acidic polysaccharide PPFS3 is extracted from an agricultural waste, namely seed pumpkin pulp, meanwhile, a fusion protein constructed by connecting a carbohydrate binding module and D-pantoic acid lactone hydrolase is provided, and the fusion protein and the seed pumpkin acidic polysaccharide PPFS3 have specific binding capacity; a simple, convenient and low-cost immobilized enzyme and recombinant protein purification method can be established. The formed immobilized fusion protein can be used for efficient and continuous production of D-pantoic acid.

The invention discloses keratinase Phyto-Keratinase and application of the keratinase Phyto-Keratinase. The keratinase Phyto-Keratinase comprises the following components: (1) protease which is coded by a nucleotide sequence as shown in SEQ ID NO.1; and / or (2) protease which is derived from bacillus subtilis, is coded by a nucleic acid sequence with more than 98% of identity with the SEQ ID NO.1 nucleotide sequence and has the characteristic of hydrolyzing keratin. According to the plant keratinase Phyto-Keratinase disclosed by the invention, the Phyto-Keratinase (plant keratinase) produced by utilizing microbial fermentation is a serine proteolytic enzyme, is also a biocompatible cutinase, and is obtained by fermenting saccharomyces cerevisiae. Phyto-Keratinase provides a nitrogen source for the growth of plants (beans), and contains various amino acids (functional peptides) and special proteolytic enzymes derived from the plants at the same time. The Phyto-keratinase can provide the effects of resisting inflammation, removing cutin and providing nutrition for skin regeneration at the same time, can relieve various inflammations by inhibiting PAR-2 receptors, and has potential value in the fields of cosmetics, beauty medicine and environment.

The invention discloses a method for inducing plant disease resistance by using a hydrolase active mutant of xyloglucan specific glucanase XEG1. A pichia pastoris eukaryotic expression system is utilized to obtain the purified hydrolase activity mutant XEG1E136D protein of the phytophthora sojae core pathogenic factor xyloglucan specific glucanase XEG1, and the hydrolase activity mutant XEG1E136D protein does not have the hydrolase activity of plant cell walls and can activate the systemic resistance of plants. The XEG1E136D is overexpressed into a biocontrol bacterium trichoderma harzianum by utilizing an agrobacterium-mediated fungal transformation optimization system, and the XEG1E136D is proved to be capable of secreting XEG1E136D protein to the outside of cells and activating the broad-spectrum systemic resistance of plants. The XEG1E136D protein expressed by the trichoderma harzianum can be used as a plant vaccine to improve the prevention and treatment effect, and the trichoderma harzianum XEG1E136D protein has a wide application prospect in the field of plant disease prevention and treatment.

The present invention relates to novel carbamate hydrolases for the enzymatic decomposition of polyurethanes and to an enzymatic process for the complete decomposition of polyurethanes into defined monomers.

The invention relates to bacillus subtilis Y32, and the preservation number of the bacillus subtilis Y32 is CGMCC (China General Microbiological Culture Collection Center) No.34819. The strain can produce xylanase with high temperature resistance and high stability, various hydrolases such as beta-mannase, protease and the like, exopolysaccharides and botrytis cinerea inhibition, and is resistant to acid, swine bile salt and 10% NaCl. The optimum temperature of xylanase produced by the strain is 55 DEG C, the optimum pH value is 6.0, under the conditions, the half-life period is 12.5 h, the relative enzyme activity is still kept above 80% after incubation for 3 h, and the xylanase has good thermal stability. The strain can also improve the yield of cucumbers and pinellia ternate, and can also improve the laying rate of 70-week-old hyline brown laying hens from 70.4% to 90.1%. The strain has a good application prospect in the fields of agriculture, food processing and cosmetic production.

The invention provides a preparation method and application of a bacterium-enzyme-magnetic nano-cluster composite detoxicating agent, the detoxicating agent takes a Fe3O4-HAP nano-cluster as a magnetic core, the surface of the detoxicating agent is subjected to functional modification, a composite flora composed of bacillus amyloliquefaciens, lactobacillus plantarum and saccharomyces cerevisiae is loaded to form sclerotia, a chitosan shell layer is constructed on the outermost layer, and the bacterium-enzyme-magnetic nano-cluster composite detoxicating agent is prepared. And a compound enzyme consisting of aflatoxin Bdegrading enzyme, zearalenone hydrolase and deoxynivalenol invertase is immobilized. The carrier can efficiently and synchronously degrade aflatoxin, vomitoxin and zearalenone, and can realize rapid separation and recovery through an external magnetic field.

The invention belongs to the field of gene engineering, and particularly relates to a PET hydrolase mutant with high catalytic activity and application thereof. According to the invention, amino acids at the 62nd site, the 65th site, the 66th site, the 71th site, the 154th site, the 156th site, the 177th site, the 183th site, the 184th site, the 209th site, the 213th site, the 214th site and the 215th site in the PES-H1L92F / Q94Y amino acid sequence of the PET hydrolase shown as SEQ ID NO.2 are mutated by utilizing a structural analysis and site-specific mutagenesis technology to obtain various mutants, and the mutants with improved catalytic activity are obtained. The PET hydrolase mutants promote the hydrolysis efficiency of ester bonds in PET so as to improve the degradation capacity of PET plastics, and can be applied to enzymatic depolymerization of PET plastics in life in the future.

The present disclosure relates to a method that includes hydrolyzing a polyamide by contacting the polyamide with a hydrolase. In some embodiments of the present disclosure, the polyamide may include nylon-6, nylon 6,6, or a combination thereof. In some embodiments of the present disclosure, the hydrolyzing may produce aminohexanoic acid, an oligomer of aminohexanoic acid, ε-caprolactam, or combinations thereof.

The application belongs to the technical field of genetic engineering, and discloses a kind of saccharomyces cerevisiae for producing rare ginsenoside by seaweed biomass and a construction method and application thereof.The saccharomyces cerevisiae has the following characteristics: overexpression of agarase, neojuncanohydrolase, hydroxymethylglutaryl coenzyme A reductase, isopentenyl diphosphate delta isomerase, dammaradienol synthase, protopanaxadiol synthase, cytochrome P450 reductase, protopanaxatriol synthase and glycosyltransferase.The application combines enzymolysis of seaweed biomass with fermentation of rare ginsenoside, which not only endows wild-type yeast with the ability to degrade seaweed biomass that it originally does not have, but also effectively improves the yield of squalene and downstream terpenes (rare ginsenoside Rh1) by overexpression of tHMG1 and IDI1.The saccharomyces cerevisiae can effectively utilize seaweed biomass to obtain squalene and rare ginsenoside Rh1, and has the characteristics of simplicity, economy and ecological friendliness, can convert cheap biomass into high-value products, and provides a way for the development of marine bioeconomy.

Aspects of the disclosure relate to Botulinum toxin X (BoNT X) protein variants. The variants provided herein have been evolved to cleave GTP cyclohydrolase 1 (GCH1). Some of the variants provided herein were evolved from a procaspase-1 cleaving polypeptide. Further aspects of the disclosure relate to nucleic acids encoding the GCH1 cleaving polypeptides described herein and expression vectors comprising the nucleic acids, as well as host cells and fusion proteins comprising the GCH1 cleaving polypeptides described herein, and kits comprising the GCH1 polypeptides, fusion proteins, nucleic acids, expression vectors, or host cells described herein. Further aspects of the disclosure relate to methods of producing BoNT X variants and methods of using the BoNT X protein variants, for example, to reduce pain.

The present invention relates to a process for the preparation of substituted 4-(N′-hydroxy-carbamimidoyl)benzoic acids, which can be obtained by nitrilase catalyzed hydration of substituted terephthalonitriles of formula (II) in an aqueous medium to afford (ammonium) 4-cyanobenzoic acids (IIa). The hydration is followed by treatment of the aqueous reaction medium with hydroxylamine or a salt thereof to afford amidoximes (I).

The invention discloses bacillus velezensis B127 and application thereof, the bacillus velezensis is preserved in China Center for Type Microbiological Culture Collection, the preservation number is CCTCC NO: M 20252878, the preservation date is December 15, 2025, the classification name is bacillus velezensis B127, and the preservation address is Wuhan University, Hubei Province. The bacillus velezensis B127 has a remarkable inhibition effect on various tobacco fusarium pathogenic bacteria, has good phosphorus and potassium solubilizing capacity, generates hydrolase related to inhibition of pathogenic bacteria and generates siderophore and indoleacetic acid for promoting plant growth, is a biocontrol bacterium with excellent comprehensive performance, and can be widely applied to the field of biological control. The method has good development and application prospects in the fields of agricultural disease control, soil improvement, microbial fertilizers and the like.

The invention discloses an SAM hydrolase mutant and application thereof, the specific activation effect of halogen anions (F <->, Cl <->, Br <-> and I <->) on SAM hydrolase (containing MJ1651 wild type and mutant) is found for the first time, and the active sequence of the SAM hydrolase is F <-> > I <-> > Br <-> = Cl <->. According to the method, 400 mM of halide salt is added into a reaction system, so that the catalytic efficiency is improved in a breakthrough manner; the yield of wild type enzyme adenosine is improved by 20-300% (optimally, an F-system); the yield of the mutant R89C is soaring up by nearly 6 times under the action of KF; the activity of the immobilized enzyme is still more than 90% after the immobilized enzyme is repeatedly used for 5 times. The method is easy and convenient to operate, green production is achieved with water as the solvent (three wastes are reduced by 90%), the comprehensive cost is reduced by 41%-80 / kg, and an efficient solution is provided for industrial biological manufacturing of antiviral drug intermediates and the like.

The invention discloses glucoside hydrolase with anti-biofilm activity and application of the glucoside hydrolase. Experiments prove that the PgaBAb1 protein and the PgaBKp protein have remarkable anti-staphylococcus epidermidis biofilm activity and can destroy a mature staphylococcus epidermidis biofilm and inhibit formation of the staphylococcus epidermidis biofilm, that is, the PgaBAb1 protein and the PgaBKp protein are glucoside hydrolase with anti-biofilm activity; meanwhile, the sensitivity of the staphylococcus epidermidis to methicillin can be improved; the amino acid sequences of the PgaBAb1 protein and the PgaBKp protein are respectively as shown in SEQ ID NO: 5 and SEQ ID NO: 6. Therefore, both the PgaBAb1 protein and the PgaBKp protein can be used as active ingredients of drugs to treat diseases caused by infection of bacteria generating biological membranes and / or drug-resistant bacteria. The method has an important application value.

The invention relates to the technical field of biology, in particular to application of glucoside hydrolase FsGH28c and a coding gene thereof in regulation of strain pathogenicity. The invention provides a glucoside hydrolase coding gene FsGH28c related to development, carbon source utilization and pathogenicity of fusarium solani, and after the FsGH28c gene is knocked out, the sporulation quantity is reduced, spore development is deformed, and the utilization rate of a carbon source (especially pectin) is reduced. In addition, the colonization amount of the FsGH28c gene deletion mutant in host pepper root fungi is reduced, and the pathogenicity is reduced. The FsGH28c gene can positively regulate the pathogenicity of fusarium solani, and can be used as an important candidate gene for research on the pathogenesis of fusarium solani.

The invention discloses a recombinant halomonas for producing hydroxyectoine as well as a preparation method and application of the recombinant halomonas. The recombinant halomonas disclosed by the invention does not contain an ectoine hydrolase (DoeA) gene, and can be used for expressing ectoine hydroxylase and glutamate dehydrogenase. Experiments prove that when the salt concentration of the recombinant halomonas is reduced to 1.4 M, fermentation can produce 1.02 g / L of hydroxyectoine, and compared with wild halomonas, the yield of hydroxyectoine under the salt concentration of 1.4 M is increased by 10 times. According to the recombinant halomonas disclosed by the invention, the hydroxyectoine is successfully produced under the condition that the salt concentration is reduced, chassis bacteria for efficiently producing the hydroxyectoine under the medium-salt condition are successfully constructed, and theoretical guidance is provided for further reducing the salt concentration for producing the hydroxyectoine; meanwhile, an experimental basis is provided for improving the biosynthesis of a high-added-value chemical hydroxyl ectoine.

The invention discloses an IV-type collagen detection kit and a preparation method thereof, and relates to the technical field of in vitro diagnos.The kit comprises a reagent R1 and a reagent R2, and the volume ratio of the reagent R1 to the reagent R2 is 3: 1; through an elaborately designed reagent formula, especially the accurate proportion of all components in the reagent R1, such as ammonium chloride, sodium azide and Brij-35, the detection sensitivity and accuracy are improved, the stability and repeatability of the kit are also ensured, and particularly, the natural plant extract astragalus polysaccharide and the protease K inhibitor are introduced into the reagent R1 as key components, so that the detection sensitivity and accuracy are improved, and the detection sensitivity and repeatability are improved. According to the present invention, the unique preparation method and the activity promotion effect on the IV-type collagen hydrolase significantly enhance the biological activity of the kit, such that the detection result is reliable, and the two latex particles coated with the paired mouse monoclonal antibodies adopted in the reagent R2 have high specificity and affinity, such that the kit can be used for the detection of the IV-type collagen hydrolase. And the detection accuracy and sensitivity are further improved.