123 results about "Chitinase" patented technology

The invention discloses an antibacterial peptide fragment intercepted from chitinase IDGF4 protein and application of the antibacterial peptide fragment in preparation of antibacterial drugs. According to the invention, short peptide fragments in chitinase IDGF4 protein are screened, three candidate peptides are obtained through chemical synthesis, the antibacterial performance of the candidate peptides is systematically evaluated, and the evaluation result shows that the peptide fragment with the amino acid sequence as shown in SEQ ID No.2 can effectively inhibit the growth of salmonella and staphylococcus aureus under the concentration of 1mg / mL; the antibacterial effect of the compound is confirmed in inhibition zone experiments, enzyme-labeled turbidimetry and growth curve monitoring, and the compound shows stable and remarkable antibacterial activity and can be used as a novel antibacterial molecule; the invention provides a new candidate sequence for developing the antibacterial peptide with high efficiency and low drug resistance risk, and has application potential in the aspect of preparing clinical antibacterial drugs.

The invention provides a high-activity chitinase mutant and application thereof in degradation of crustacean waste, and belongs to the technical field of enzyme engineering and agricultural waste conversion. According to the invention, chitinase PbChi from Paenibacillus barengltzii is subjected to molecular modification, and a plurality of chitinase mutants with high activity are successfully constructed by increasing the flexibility of a connexin, changing steric hindrance at a substrate catalytic crack inlet and changing catalytic residues in a beta sheet layer in a catalytic structural domain through calculation design and experimental verification; the chitinase mutant can effectively degrade shrimp shell meal, so that carapace waste is effectively utilized, and the chitinase mutant has a wide application prospect in the biological feed industry.

The invention discloses application of oilseed rape chitinase EP3 and a coding gene thereof in improving clubroot resistance of cruciferous plants, and belongs to the technical field of molecular biology. The invention discloses the application of the rape chitinase EP3 in improving the clubroot resistance of plants for the first time. The amino acid sequence of the rape chitinase EP3 is as shown in SEQ ID NO.2; the nucleotide sequence of the coding gene EP3 is shown as SEQ ID NO. 1. Experimental results show that by preparing agrobacterium containing an EP3 gene overexpression vector and using a treating fluid containing the agrobacterium to perform seed soaking and root irrigation treatment, the EP3 gene is overexpressed in a rape plant, and the clubroot resistance of the rape can be remarkably improved. The invention provides a new gene resource and a safe, efficient, simple, convenient and universal prevention and treatment method for preventing and treating clubroot of cruciferous plants, and has remarkable agricultural application value and wide application prospect.

The invention belongs to the technical field of chitosan oligosaccharide preparation, and particularly relates to a method for preparing chitosan oligosaccharide based on compound enzyme. According to the method for preparing the chitosan oligosaccharide based on the compound enzyme, a multi-enzyme synergistic system is matched with the design of a specific protective agent, chitosanase and chitinase can cooperatively cut beta-1, 4 glycosidic bonds of chitosan and control molecular weight distribution of products, meanwhile, chitinase can directionally cut the beta-1, 4 bonds at the tail end of chitosan, and the molecular weight distribution of the products is controlled. The papain is used for removing protein impurities in the chitosan to realize accurate control of the polymerization degree. Besides, a specific protective agent matched with the multi-enzyme synergistic system is specifically designed, the specific protective agent and the multi-enzyme composition act together, the molecular weight distribution width can be remarkably reduced, the yield and the DP 4-8 proportion can be increased, meanwhile, physical field ultrasonic assistance is added in the enzymolysis treatment process, and the enzymolysis treatment efficiency is improved. The combination of the substrate and the enzyme is further promoted, and the reaction time is shortened.

The invention relates to the technical field of agriculture, and discloses a synergistic technology for repairing soil and promoting plant stem cells by using modified amino active groups. Comprising the following steps: modifying an amino acid compound, namely modifying active groups in amino acid and fulvic acid, so that the active groups have a synergistic effect with cellulase, hemicellulase, urease, phosphatase, chitinase, beta-glucanase, superoxide dismutase, peroxidase and catalase; through the strong chelation of the modified amino active groups and heavy metal ions in soil, the bio-availability and mobility of heavy metals can be effectively reduced, for example, the chelation removal rate of cadmium, lead and other heavy metals can reach 40% or above, the heavy metal contaminated soil is repaired, the soil structure is improved, the content of organic matters in the soil is increased, and the pH value of the soil is adjusted; the acidified or alkalized soil is gradually restored to a pH range suitable for plant growth, and the water and fertilizer retention capability of the soil is improved.

The invention belongs to the technical field of agricultural fertilizers, and particularly relates to a high-efficiency yield-increasing liquid fertilizer containing enzymolysis nereis and a preparation method thereof.The method comprises the steps that the nereis is cleaned and smashed, subjected to freeze-thaw cycle pretreatment and then subjected to enzymolysis through a compound protease preparation containing protease and chitinase under the alkalescence condition, and the liquid fertilizer containing the enzymolysis nereis and the chitinase is obtained; carrying out solid-liquid separation on the obtained enzymatic hydrolysate to obtain enzymatic clam worm liquid; the preparation method comprises the following steps: mixing urea, a phosphorus-potassium fertilizer, trace elements, potassium fulvate and polyaspartic acid in a preparation tank, and then adding an N-carbamoyl-L-glutamic acid-chitosan grafted copolymer and selenocysteine-alginic acid nanoparticles to obtain a nutrient solution; and finally, mixing the enzymolysis nereis solution with the nutrient solution and the plant growth regulator to obtain a finished product. Active substances in the nereis are fully released through a biological enzymolysis technology, the prepared liquid fertilizer has the functions of nutrition supply, physiological regulation and soil improvement, crop growth can be remarkably promoted, the yield can be remarkably increased, and high-value utilization of marine biological resources is achieved.

The invention discloses a Chinese herbal medicine leavening, a preparation method thereof and application of the Chinese herbal medicine leavening in cosmetics, and particularly relates to the technical field of cosmetics. Comprising the following raw materials in parts by weight: 12-18 parts of radix astragali, 8-12 parts of radix ampelopsis, 10-15 parts of radix saposhnikoviae, 18-22 parts of rhizoma chuanxiong, 6-9 parts of bitter almond seeds, 18-22 parts of radix salviae miltiorrhizae, 7-10 parts of herba centellae, 6-9 parts of folium artemisiae argyi, 7-10 parts of fructus gardeniae, 4-6 parts of flos sophorae immaturus, 5-8 parts of fructus citri, 12-15 parts of poria cocos and 1.5-2.5 parts of a dendrobium officinale extract. The composition can synergistically exert moisturizing, immune regulation and anti-allergy effects with other raw material components to form an anti-allergy, anti-oxidation and repair efficacy closed loop, a process chain from aspergillus oryzae high-enzyme-activity solid fermentation to double-bacterium liquid symbiosis to chitinase directional enzymolysis can synergistically improve the yield of active matters, and the situation that traditional anti-allergy components have no hair growth effect is broken through, so that the anti-allergy and anti-allergy effects are achieved. Due to the limitation that the hair restorer stimulates sensitive skin, the hair growing and double effects are integrated, and the hair restorer is suitable for preparing cosmetics for resisting allergy, resisting oxidation and repairing barriers and is especially suitable for sensitive skin care and scalp care.

The invention discloses a novel brevibacterium root rot resistant species of Huanglonglong and application of the novel brevibacterium root rot resistant species. The invention provides a Huanglonglong brachybacterium root rot resistant strain, which is a Huanglonglong brachybacterium root rot resistant strain, the strain number of the Huanglonglong brachybacterium root rot resistant strain is BS409, and the registration number of the Huanglonglong brachybacterium root rot resistant strain in the China General Microbiological Culture Collection Center is CGMCC No.34201. The invention further provides a preparation method of the Huanglonglong brachybacterium root rot resistant strain. The strain BS409 provided by the invention is a new species of brevibacterium, can generate chitinase and hydrolyze chitin, and has inhibitory activity on root rot pathogenic bacteria fusarium equiseti (Fusarium equiseti). The results indicate that the strain BS409 has good potential in the aspect of biological prevention and control of root rot of medicinal plants (such as bighead atractylodes rhizome, pseudo-ginseng and ginseng), and can also be used in the aspects of soil remediation, soil improvement and the like of planting fields of bighead atractylodes rhizome, pseudo-ginseng and ginseng.

The present application relates to "application of a compound in soybean cyst nematode chitinase inhibitor", belong to the technical field of leguminous plant symbiotic protection, the application of the compound and its pharmaceutically acceptable salt in soybean cyst nematode chitinase inhibitor, the present application is evaluated and researched through the inhibitory activity of compound by screening work, the inhibitor is screened finally, the inhibitor is likely to have good application prospect in controlling soybean cyst nematode disease aspect.The present application also provides a kind of chitinase inhibitor with the function of inhibiting the establishment of symbiosis of soybean cyst nematode to destroy, the number of nodule obtained by processing is flat with the number when only inoculating rhizobium, the inhibitory effect of Hg-CHI-1 on rhizobium nodule is completely restored, and it has good application prospect in the aspect of leguminous plant symbiotic protection.

This invention discloses a chitinase-producing Bacillus albus A01 and its application in the fermentation of shrimp and crab solid waste. The chitinase-producing Bacillus albus is named... Bacillus albus A01, with accession number CCTCC: M 20252571; the 16S rDNA gene sequence of this chitinase-producing Bacillus albus strain A01 is shown in SEQ ID NO.1. The fermentation agent of this strain can effectively degrade the chitin content in shrimp and crab solid waste, and improve the nutritional value of shrimp and crab fermented powder through solid-state fermentation. This invention solves the problem of unsatisfactory fermentation effects of existing feed fermentation strains, and has great application potential for improving the feed nutritional quality of shrimp and crab fermented powder and developing feed resources.

The present application relates to a kind of organic water-soluble fertilizer and its preparation method, which uses fish waste and shell waste as raw material, after being crushed and sterilized, enzymatic hydrolysis is carried out using acid protease and chitinase to obtain enzymatic hydrolysate;Add molasses to the enzymatic hydrolysate, and inoculate mixed bacterial agent for primary fermentation to obtain fermentation liquor A;Add yeast, neutral protease and sodium alginate to the fermentation liquor A, and carry out secondary fermentation under stirring to obtain fermentation liquor B;After treating the fermentation liquor B, add chitosan to obtain the organic water-soluble fertilizer;The organic fertilizer prepared by the method of the present application can utilize shell waste, and the obtained organic water-soluble fertilizer can improve its water-retaining and fertilizer-retaining performance in sandy soil while meeting the standard requirements of amino acid water-soluble fertilizer.

The application belongs to the technical field of agricultural fertilizer, and particularly relates to a high-efficiency yield-increasing liquid fertilizer containing enzymatic nereid and a preparation method thereof, which comprises the following steps: washing and crushing nereid, and then performing freeze-thaw cycle pretreatment; under weak alkaline conditions, a complex protease preparation containing protease and chitinase is used for enzymolysis, and an enzymatic nereid liquid is obtained through solid-liquid separation; urea, phosphate-potassium fertilizer, trace elements, potassium fulvate and polyaspartic acid are mixed in a preparation tank, and then N-carbamoyl-L-glutamic acid-chitosan graft copolymer and seleno-cysteine-alginic acid nanometer microspheres are added to obtain a nutrient liquid; finally, the enzymatic nereid liquid, the nutrient liquid and a plant growth regulator are mixed to obtain a finished product. Through the biological enzymolysis technology, the active substances in the nereid are fully released, the prepared liquid fertilizer has the functions of nutrient supply, physiological regulation and soil improvement, can significantly promote crop growth and increase yield, and realizes high-value utilization of marine biological resources.

The invention relates to construction and application of an engineering bacterium for high yield of chitinase ChiD, in particular to a bacillus thuringiensis engineering bacterium deltacdsR (P5014-chiD) strain, and the preservation number of the bacillus thuringiensis engineering bacterium deltacdsR (P5014-chiD) strain is GGMCC No.35124. The HD73 endogenous gene chiD gene is efficiently expressed in the bacillus thuringiensis delta cdsR, and the yield in a fermentation culture medium is far higher than that of a wild strain HD73. Compared with HD73 (P5014-chiD), the compound has a relatively strong inhibition effect on mycelial growth of fusarium graminearum and exserohilum turcicum, and shows a better effect on prevention and treatment of larvae of plutella xylostella compared with the HD73 (P5014-chiD).

The invention provides a method for preparing micromolecular glucosamine peptide by utilizing leftover materials of aquatic products. The method comprises the following three steps: firstly, treating raw materials through alkaline hydrolysis, and removing impurities; secondly, chitinase, deacetylase and collagenase are added at a certain temperature for enzymolysis, and the raw materials are converted into micromolecular collagen peptide; and finally, carrying out activated carbon adsorption, ultrafiltration membrane filtration and heating evaporation concentration to obtain a high-purity glucosamine peptide solution. The method is efficient, environment-friendly and low in cost, waste generated in the aquatic product processing process can be fully utilized and converted into micromolecular amino glycopeptides with commercial value, cyclic utilization of resources is promoted, and the method has high sustainable development significance and wide industrial application prospects.

The invention discloses a high-deacetylation-degree chitosan preparation method, which comprises: suspending chitin in a solvent to prepare a chitin suspension; adding a compound enzyme into the obtained suspension liquid to carry out enzymatic reaction, so as to prepare the chitosan with high deacetylation degree; wherein the compound enzyme is prepared from chitin deacetylase, chitinase, beta-N-acetylglucosaminidase and protease. According to the method, four enzymes with complementary functions are creatively combined to form a unique technical path of four-stage synergistic interaction of'removal-deconstruction-assistant-deacetylation ', so that a natural chitin crystal barrier can be efficiently deconstructed, the protein shielding effect is overcome, and the effect of directly preparing high-deacetylation-degree chitosan from chitin is achieved.

The application discloses a kind of low temperature resistant strain YW-1 for preventing and treating rice seedling wilt, and belongs to the field of plant protection.The low temperature resistant strain YW-1 is a pseudo-odor class fungus, and the application proves that the strain YW-1 can maintain normal growth and metabolism in a low temperature environment of 15 DEG C, and can effectively destroy the hyphal structure of the wilt pathogen by secreting cellulase and chitinase, thereby inhibiting the occurrence of the disease from the source.The strain YW-1 has no antagonism with the commonly used functional fungus Trichoderma in the seedling raising substrate, and the two can be compounded and coexist, thereby realizing the synergistic effect of functional microorganisms.The application overcomes the defects of the existing biocontrol strain, such as reduced activity in a low temperature environment and unstable prevention effect, and provides a stable and reliable biological control scheme for factory seedling raising in a low temperature season without increasing the amount of chemical pesticides, thereby significantly improving the quality of seedlings and the seedling raising success rate.

The present invention relates to a composition for bone repair and growth and a preparation method thereof, belonging to the field of medical technology. The composition includes a peptide extracted from the gelatinous layer of sand jellyfish and a bamboo yellow-Grifola frondosa composite bacterial peptide; the peptide extracted from the gelatinous layer of sand jellyfish contains a product between 500Da and 2000Da obtained by sequentially hydrolyzing the gelatinous layer of sand jellyfish with lumbrokinase, serrapeptase, and transglutaminase; the bamboo yellow-Grifola frondosa composite bacterial peptide contains a product between 500Da and 2000Da obtained by co-enzymatic hydrolysis of bamboo yellow mushrooms and Grifola frondosa mushrooms with cellulase and chitinase, then co-enzymatic hydrolysis with nattokinase and serrapeptase, and finally co-enzymatic hydrolysis with aminopeptidase and transglutaminase. The present invention adopts a special method to prepare the peptide extracted from the gelatinous layer of sand jellyfish and the bamboo yellow-Grifola frondosa composite bacterial peptide, and the combined use achieves good biological activity, stability, and absorption utilization rate, overcomes the defects of existing bone repair and growth drugs, and effectively promotes bone repair and growth.

This invention discloses a method for improving the resistance to postharvest black spot disease in sweet potato tubers and maintaining their quality, belonging to the field of postharvest preservation technology for agricultural products. The method includes: selecting undamaged sweet potato tubers for callus treatment at room temperature for 48 hours, disinfecting with 1% sodium hypochlorite, rinsing with sterile water and air-drying, then soaking them in a 9.0 mmol / L benzothiadiazole (BTH) solution (containing 0.05% Tween 80) at room temperature for 60 minutes, removing them and air-drying them to complete the treatment. This invention induces the activation of the sweet potato tuber's own defense system through BTH, increasing the activity of key enzymes in phenylpropane metabolism, chitinase and other disease-resistant enzymes, promoting the accumulation of total phenols and lignin, maintaining cell membrane integrity, inhibiting starch degradation and abnormal increase in soluble sugars. Under storage conditions of 28±1℃ and 80-85%RH, it can significantly inhibit the expansion of black spot disease lesions and extend the storage period to more than 25 days. This method has no pesticide residues, is unlikely to cause pathogen resistance, is environmentally friendly, simple to operate, and cost-controllable, and is suitable for large-scale postharvest preservation of sweet potato tubers.

The invention relates to the technical field of gene engineering, and particularly discloses a chitinase truncated body, a recombinant expression vector thereof, and a construction method and application of engineering bacteria. The chitinase truncated body is Cq181NC, the amino acid sequence of the Cq181NC is as shown in SEQ ID NO: 1, and the nucleotide sequence for coding the chitinase is as shown in SEQ ID NO: 2. According to determination, the chitinase truncation provided by the invention has the advantages that the specific enzyme activity is obviously improved by 1.8 times compared with that of a wild type, the chitinase truncation is high in environmental change tolerance, and the thermal stability of the chitinase truncation is still 1.43 times that of the wild type (incubation is performed for 8 hours at 35 DEG C). More importantly, the spectrum of the enzymolysis product is changed: the wild type is mainly disaccharide and shows a typical excision enzyme mode; and the truncated Cq181NC generates a plurality of oligosaccharides including monosaccharide, disaccharide, trisaccharide and hexasaccharide, shows double catalytic characteristics of endonuclease and exonuclease, and has a better application prospect.

The invention discloses a method for producing oligosaccharide by cooperatively treating aspergillus niger residues through multiple enzymes. The method comprises the following steps: mixing pretreated aspergillus niger residues with hydrochloric acid, carrying out acidolysis wall breaking, and carrying out suction filtration on an obtained wall-broken mixture; carrying out mixed treatment on filtrate obtained by suction filtration with cellulase and beta-glucanase, filtering, and carrying out concentration and alcohol precipitation on supernate to obtain soluble beta-glucan oligosaccharide; performing filter residue enzymolysis on filter residues obtained by suction filtration by using mesoporous silica immobilized chitinase, and recovering the immobilized chitinase after enzymolysis; and filtering the mixed solution subjected to enzymolysis by using the immobilized chitinase, and carrying out alcohol precipitation on the supernate to obtain the N-acetylglucosamine and the chitosan oligosaccharide. According to the method disclosed by the invention, the aspergillus niger residues are jointly treated by adopting multiple enzymes, the yield of N-acetylglucosamine and chitosan oligosaccharide reaches 85.1%-88.5%, and the yield of beta-glucosamine oligosaccharide reaches 86.7%-89.3%. The invention provides a brand new route for producing various oligosaccharides by utilizing solid waste aspergillus niger residues in the fermentation industry.

The invention relates to the technical field of insect resource utilization, and discloses a comprehensive utilization method for extracting protein and grease from insects, which comprises the following steps: selecting hermetia illucens as a raw material, crushing, adding water with the weight 2.5-3.5 times that of the raw material, and stirring and mixing; grinding the mixture to 40-120 microns through bone paste grinding equipment; adding chitinase, lipase and protease which account for 0.015-0.035% of the total amount, and carrying out enzymolysis at 52-58 DEG C for 3.5-6.5 hours; raising the temperature to 90-100 DEG C and carrying out enzyme deactivation for 25-35 minutes; homogenizing by a colloid mill until the particle size is 3-15 microns; solid residues, a grease phase and a liquid protein phase are separated out through a disc type centrifugal machine; carrying out spray drying on the liquid protein phase to obtain degreased insect protein powder with moisture less than 6%; and standing and layering the oil phase, and filtering with a 0.2-1.2 micron filter membrane to obtain the refined insect oil with the acid value of less than 2mgKOH / g. According to the invention, the quality and nutritional value of the protein product are improved, the equipment blockage risk and energy consumption are reduced, and the applicability of the grease food-grade standard is improved.

The invention relates to the technical field of parasite prevention and control, and provides a probiotic fungus capable of promoting parasite egg death and a preparation method thereof, the strain is recombinant aspergillus niger, and an exogenous chitinase gene (SEQ ID NO: 1) and a keratinase gene (SEQ ID NO: 2) are integrated in a genome of the strain. Aspergillus niger is transformed through genetic engineering, multi-enzyme synergistic degradation of the horny cortex of the worm eggs of the echinococcosis is achieved, the survival rate of the worm eggs is remarkably reduced, meanwhile, environmental safety and long-acting prevention and control capacity are both considered, and an innovative solution is provided for comprehensive prevention and control of echinococcosis.

The invention discloses a bacillus subtilis CR-6 and application thereof, and relates to the technical field of microorganisms, and the preservation number of the bacillus subtilis CR-6 is CGMCC (China General Microbiological Culture Collection Center) No.35148. The bacillus coagulans ER-6 has the characteristic of high yield of chitinase, has a strong contact killing effect on meloidogyne incognita and reniform reniform nematodes no matter fermentation liquor or fermentation supernate, is applied to prevention and control of agricultural pests, does not involve broad-spectrum chemical toxins, does not have harmful residues, and meets the requirements of green agriculture and ecological safety. Meanwhile, the bacillus coagulans ER-6 disclosed by the invention also has a growth promoting effect on plant growth, and the yield of crops is remarkably increased.

The invention discloses a beta-N-acetylglucosaminidase mutant obtained based on random mutation and a preparation method of the beta-N-acetylglucosaminidase mutant. On the basis of an error-prone PCR (Polymerase Chain Reaction) technology, a mutant BsNagZ-14 (C17R / K209T / T436S) and a mutant BsNagZ-17 (E358G) of the beta-N-acetylglucosaminidase of the bacillus subtilis are obtained in the invention. The two mutants keep high enzyme activity within the pH range of 3-10, the thermal stability of BsNagZ-14 is better, 98% of enzyme activity (90% of original enzyme) is kept after the temperature is kept at 40 DEG C for 3 hours, 83% of enzyme activity is kept after the temperature is kept at 60 DEG C for 3 hours, and the parent is attenuated to 23% of the initial enzyme activity. The mutants BsNagZ-14 and BsNagZ-17 are connected with a carrier pET20b (+) and are transferred into escherichia coli BL21 (DE3), the extracellular highest enzyme activities of the two recombinant bacteria are 1.140 U / mL and 1.136 U / mL respectively, and the extracellular highest enzyme activities of the two recombinant bacteria are improved by 16.32% and 15.91% respectively compared with those of control group bacteria expressing NagZ. The beta-N-acetylglucosaminidase can be externally tangent to the chitosan oligosaccharide, so that the hydrolysis effect of the chitinase on the chitin is favorably improved. The beta-N-acetylglucosaminidase mutant has wide pH and temperature adaptability, can be adapted to chitinase with different enzymatic characteristics, and provides convenience for carrying out double-enzyme hydrolysis on chitin.

The invention provides a high-activity exo-chitinase mutant and application of the high-activity exo-chitinase mutant in degradation of chitin waste, and belongs to the technical field of enzyme engineering and waste bioconversion. According to the present invention, the semi-rational engineering design is performed on the wild type exo-chitinase derived from Paenibacillus barengltzii, and a group of exo-chitinase mutants with characteristics of high activity, high stability and strong substrate binding affinity are constructed through the site-directed mutagenesis and domain fusion strategy; the exo-chitinase mutant shows excellent thermal stability and pH stability in a wide temperature and pH range, and the exo-chitinase mutant or an enzyme preparation containing the exo-chitinase mutant can be used for large-scale degradation of agricultural and marine chitin wastes. The method has a wide application prospect in the fields of sustainable feed production, agriculture and industrial biotechnology.

This invention discloses a truncated chitinase TSCHI-T, its preparation method, and its applications. The amino acid sequence of the truncated chitinase TSCHI-T is shown in SEQ ID NO.3. The truncated chitinase obtained by this invention has high hydrolytic activity for low-deacetylated chitosan. The crude enzyme solution produced by shake-flask fermentation has the ability to degrade 10g of low-deacetylated chitosan by 1mL, while hydrolyzing the same mass of low-deacetylated chitosan requires approximately 300mg of non-specific commercial enzyme. The enzymatic hydrolysis efficiency is increased by nearly 700 times, and it can be used for large-scale preparation of low-deacetylated chitosan oligosaccharides.

The invention relates to a compound enzyme composition and a preparation method of cordyceps sinensis protoplast, and belongs to the technical field of bioengineering. The compound enzyme composition is prepared from Yatase, chitinase and beta-glucanase; wherein the mass ratio of the Yatalase to the chitinase to the beta-glucanase is (1-2): (1-2): (1-3). The preparation method comprises the following steps: S1, culturing and purifying hyphae; s2, preparing a protoplast; the protoplast is prepared by mixing a compound enzyme composition with the purified cordyceps sinensis mycelia obtained in the step S1, and reacting; wherein enzymes in the compound enzyme composition comprise Yatase, chitinase, beta-glucanase and a solvent. When the compound enzyme composition provided by the invention is used for carrying out enzymolysis on the cordyceps sinensis mycelia, the obtained cordyceps sinensis protoplast is high in yield, short in time consumption, complete in form, normal in size and low in required cost, so that the research cost can be reduced, and the industrial production can be expanded.

The invention discloses a method for synchronously preparing chitosan and citric acid through a kitchen waste lobster shell fermentation method, belongs to the field of solid waste resource utilization, and aims to solve the problems of difficult treatment, low utilization rate and pollution caused by impurity wrapping in shellfish kitchen waste treatment. According to the method, kitchen waste containing grease, chitin, calcium carbonate and protein is used as a raw material, and two-stage treatment mainly including fermentation is carried out. In the first stage, citric acid is produced through fermentation, and chitin is exposed through in-situ deoiling, decalcification and deproteinization; in the second stage, enzymolysis is mainly carried out, and after the pH is adjusted, chitinase and chitin deacetylase are exogenously added to convert chitin into chitosan; and finally, separating the feed liquid to obtain citric acid and chitosan. According to the method, fermentation is combined with enzymolysis segmented treatment, so that the problem of substrate wrapping is solved, the conversion rate and purity of chitosan are remarkably improved, high-valued co-production of waste is realized, and the process is mild and environment-friendly.

The invention concerns a method for prophylaxis of infections by fungi, particularly by oomycetes, and of bacterial infections in crop and ornamental plants. Areas of application are vine, fruit, vegetable and ornamental plant growing. The method of the invention is characterized in that an aqueous solution of a protease, alone or in combination with β-glucanases and / or chitinases, is prepared, stabilizers, stickers and wetters are added, and the additized solution is applied by conventional techniques to the plants a number of times within the vegetation period, preferably ahead of weather-related phases of high infection threat.