119results about How to "High enzyme production" patented technology

The invention relates to a method for preparing algin oligosaccharide by using algin lyase Agarivorans albus YKW-34, which is characterized by comprising the following steps: firstly preparing enzyme liquid of the algin lyase Agarivorans albus YKW-34, dissolving algin in water, adding the prepared enzyme liquid into the solution, decomposing the algin for 2 to 180 hours in a water bath at a temperature of 20 to 55 DEG C, raising the temperature to between 60 and 100 DEG C, inactivating the algin, filtering the algin by a film, and condensing, freezing and drying the algin to obtain the algin oligosaccharide. The method has the advantages that a culture medium for preparing the algin lyase has economic and simple compositions; the algin lyase has high yield, unique Na<+>/K<+> ion dependence, superior temperature and pH stability and high specific activity, and can degrade seminose half aldehyde and guluronic acid fragments of the algin at the same time; and the produced algin oligosaccharide with the polymerization degree of 2 to 12 has favorable application prospect.

The invention discloses an omega-aminotransferase mutant gene and an application thereof, and belongs to the field of the molecular biological technology. The nucleotide sequence of the omega-aminotransferase mutant gene is shown as SEQ ID NO.1. The omega-aminotransferase amino acid sequence embodied by the code of the omega-aminotransferase mutant gene is shown as SEQ ID NO.2. The omega-aminotransferase mutant gene is firstly provided, and the omega- aminotransferase corresponding to the code of the omega-aminotransferase mutant gene has a high enzyme amount, so that the biological catalysis and conversion efficiency is improved in industrial production. The invention further provides the application of the omega-aminotransferase embodied by the code of the omega-aminotransferase mutant gene in preparation of trifluoro-propylamine. The trifluoro-propylamine is added to a buffer solution to serve as the substrate for the reaction, and then products of trifluoromethyl amine compounds are collected from the reaction liquid. The method for preparing the trifluoro-propylamine has the obvious advantages that the reaction conditions are mild, no pollution is caused, and the process is simple.

The present invention provides a method for directed selective solid-phase culturing of stable microbial mixed populations for the continuous preparation of defined enzyme and metabolite mixtures, enzyme and metabolite materials obtained by such method and suitable devices therefore.

The invention discloses bacillus subtilis JYM841 with antibacterial activity. The bacillus subtilis is preserved in a China center for type culture collection in January 10, 2014, and the preservation number is CCTCC NO: M 2014012. The bacillus subtilis JYM841 has a strong inhibitory effect on staphylococcus aureus and candida albicans, is high in antibacterial activity, wide in antibacterial spectrum, high in enzyme production, sensitive to bacterial antibiotic, drug-resistant to fungal antibiotic, high in heat-resisting ability and tolerant to acid and alkaline, has high antibacterial activity after treated at the temperature of 100 DEG C for 20min, and has tolerance to various enzymes such as trypsin, papain and protease K. An acid and alkali resistant pH (potential of hydrogen) value ranges from 6 to 10, and metabolite is stable, so that the bacillus subtilis can be applied to preparation of medicines for treating genital tract infection.

A genetically engineered yeast Pichia pastoris (GS115/HBMO47) able to effectively express beta-mannase is prepared through screening beta-mannase gene from the environmental microbes, cloning it to the expression carrier of Pichia yeast, introducing the carrier to Pichia yeast, and screening. The process for preparing the high-activity beta-mannase and the high-purity oligomannosan from said Pichia pastoris is also disclosed.

A screening and cultivation method for Bacillus thuringiensis of high-yield high temperature resisting protease includes the following steps: high-yield high temperature resisting protease Bacillus thuringiensis Bt140 is obtained through adopting casein substrate agar to identify flat primary screen and fermenting enzyme activity measuring secondary screening; Plackett-Burman design and response surface analytical method are adopted to obtain optimal fermentation zymogenic formula and fermentation condition for Bt140 quickly; finally, shaking fermentation level reaches to 918.56U/mL. During adopting Bt140 to produce high temperature resisting protease, spore collection for producing pesticide after fermentation is not influenced; therefore, the Bacillus thuringiensis combines the production capacity of protease and pesticide. For example, when 1 ton Bacillus thuringiensis is used for fermentation, corresponding Bt pesticide is obtained along with 60 to 90 Kg/ megaunit enzyme powder.

The method for preparing soybean isoflavone aglycone by using enzyme method to hydrolyze soybean isoflavone glycoside glycosyl includes the following steps: using the enzyme capable of hydrolyzing soybean isoflavone glycoside glycosyl, mixing said enzyme, soybean isoflavone glycoside and buffer solution to make reaction for 1-40 hr., its reaction temp. is 4-75 deg.C and pH value is 3-9, so as to extract soybean isoflavone aglycone. Said invention not only can use enzyme to hydrolyze monoglycoside-soybean glycoside and genistein glycoside and convert them into correspondent aglycone, but also can use enzyme to hydrolyze soybean glycoside and genistein glycoside mixed glycoside and convent them into the mixed aglycone. Said product can be used as raw material for making medicine and health-care product.

A process for degradating feather by use of microbes includes such steps as preparing the substrate of feather to be transformed, obtaining the Bacillus licheniformis ZJUEL31410 (CGMCC 1397), and microbial transforming of the feather to obtain forage.

The invention belongs to the preparation of a new gene, and in particular discloses a glucose oxidase gene GOD, protein coded by the GOD and pichia pastoris transformed by the GOD. The glucose oxidase gene GOD is prepared by taking 5'-ATGAAGTCCACTATTATCACCTCCA and 3'-CTAGGCACTTTTGGCATAGTCTTCA as specific primers and penicillium notatum as a template through polymerase chain reaction (PCR) amplification. According to the invention, the problem of an existing recombinant glucose oxidase gene in heterologous expression is solved; the total-length GOD gene and a shuttle expression vector constructed by the gene are obtained and the gene and the shuttle expression vector thereof are further transformed to the pichia pastoris, and upon screening and identification, a strain, which is higher in secretion expression of the glucose oxidase than an original strain, is obtained. The invention can greatly save cost and expense for further industrialized production and can improve the economic benefits of the glucose oxidase.

The invention discloses a feed enzymic preparation prepared by using soy sauce residues. The feed enzymic preparation is prepared by adopting a preparation method comprising the following steps of: (1) preparing materials: taking desalted and dewaterd soy sauce residues as the main raw materials of a culture medium, adding inorganic salts into the culture medium, and adding water and evenly mixing to form a zymogenic material, wherein the dosage of the added inorganic salts is 0-0.25% of the total dry weight of the culture medium; (2) steaming: steaming the zymogenic material; (3) inoculating: inoculating strains into the steamed zymogenic material, and evenly mixing to form a fermenting material; and (4) fermenting: fermenting the fermenting material at the temperature of 28-35 DEG C until the fermenting material is mature, so as to obtain the feed enzymic preparation. The feed enzymic preparation has the advantages of higher acid protease activity, glucoamylase activity and cellulase activity and is suitable for being used as an animal feed. The invention further discloses the preparation method of the feed enzymic preparation.

The invention discloses acetaldehyde dehydrogenase and an application thereof in an aspect of anti-alcoholic health-care products, and belongs to the technical field of enzyme engineering. The invention provides an engineering bacterium for high-efficiency expression of a human acetaldehyde dehydrogenase gene and a construction method of the engineering bacterium; the expression of the humanized acetaldehyde dehydrogenase gene, which is shown as SEQ ID NO.1, in bacillus subtilis is achieved; and an enzyme yield by fermenting the recombinant bacterium for 24h can reach 24.52U/L or above. By improving a fermentation medium, the yield of the acetaldehyde dehydrogenase is improved, and an enzyme yield of a culture medium which is added with CaCl2 salt and CuSO4 is improved to 140-180% in comparison with an original enzyme yield; therefore, a new way and a theoretical basis are provided for research and development of anti-alcoholic drugs.

The invention relates to Bacillus amyloliquefaciens and a method for producing chymosin by fermenting using the same and belongs to the technical field of microbial fermentation. The invention discloses a bacterial strain for producing the chymosin. The bacterial strain is named as Bacillus amyloliquefaciens JUN002 and is collected in China Center for Type Culture Collection, wherein the collection number is CCTCC No:M2011045. The method for producing the chymosin by using the bacterial strain through liquid fermentation comprises the following steps of: performing slant culture; (2) performing seed culture; and (3) performing liquid fermentation culture, namely inoculating cultured seeds into a fermentation culture medium according to the inoculation quantity of 0.1 to 0.2 percent to culture the seeds at the temperature of between 32 and 36 DEG C for 60 to 72 hours. The Bacillus amyloliquefaciens and the method for producing the chymosin by fermenting using the same have the advantages that: the chymosin is produced by using the Bacillus amyloliquefaciens CCTCC No:M2011045 through liquid fermentation, so the enzyme production level is high, the fermenting time is short, growth conditions are easy to control and the like; and the method is completely suitable for application to industrialized production.

The invention provides acid protease which is derived from microorganism sources and a preparation method thereof. The acid protease is characterized in that a. enzymology characteristics: the optimumpH is 2.5-3.5, the stable pH is 2.5-6.0, the optimum temperature is 40-50 DEG C, and the temperature stability ranges from 30 DEG C to 50 DEG C; b. with A. niger as an enzyme producing strain and wheat bran as main materials, the acid protease is prepared by solid fermentation process; the fermenting and enzyme producing capabilities are more than or equal to 47000u/g (based on dry yeast), the yield of liquid enzyme is more than or equal to 85% and that of solid enzyme is more than or equal to 80%, which all reach the food grade hygienic standards. The acid protease takes grain processing by-products as main raw materials and has high fermentation enzyme activity, high enzyme extraction purification yield and low production cost. The acid protease is suitable for serving as the preparation raw material or the additive of the products in such industries as tanning, medicine, brewing and feed.

This invention is with the guide hammer bacterium of culture medium preparation Lin the method of sour glycerine oxidize enzyme. The group of culture medium divides and cultivates condition: Bevel guides culture medium, draw thing by yeast , egg white Dong , glucose , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, agars, the magnesiums of sulphuric acid, chlorination manganese, common salts and pH6.8s forms; Seed culture medium draws thing and egg white by yeast Dong , glucose , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, the magnesiums of sulphuric acid, chlorination manganeses, common salts and pH6.8s forms; Seed culture medium draws thing and egg white by yeast Dong , glucose , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, the magnesiums of sulphuric acid, chlorination manganeses, common salts and pH6.8s forms; Ferment culture medium draws thing and egg white by yeast Dong , Lin the mixture salt solution of 2 sour hydrogen potassiums, glycerines, the magnesiums of sulphuric acid, chlorination manganeses, common salts and pH6.8s forms. Bacterial strain is cultivated by bevel, seed is cultivated , revolving shaking table ferments to train , enlarging again collect bacterium body after training , adoption assembles glycol - the two water of ammonium sulphate appearance extraction and the layer of two steps Xi purification gets Lin sour glycerine oxidize enzyme.

The invention provides cerrena unicolor derived laccase and a gene and application thereof, and belongs to the field of biotechnology. The amino acid sequence of the cerrena unicolor derived LacB laccase provided by the invention is as shown in SEQID NO.3, the overall-length sequence of the gene of the cerrena unicolor derived LacB laccase is as shown in SEQID NO.1, and the cDNA sequence is as shown in SEQID NO.2. The invention further discloses a fermentation production method of the LacB laccase. The LacB laccase is produced through fermentation of the cerrena unicolor, after the sixth day of fermentation culture, the laccase yield can reach 652 U/mL, and the fermentation production method has the advantages of being high in yield and short in enzyme production cycle. The LacB laccase has significant effects in environmental hormone degradation, and the degradation rate on diethylstilbestrol, hexestrol, dienestrol, beta-estradiol, bisphenol a and hydroprednisone is 98% or above. TheLacB laccase provides a new way for environmental pollutant government, and has good economic benefits and social benefits.

The invention discloses an application of Haloferula sp. beta-N-acetylhexosaminidase to synthesis of breast milk oligosaccharide. The invention provides an application of HaHex74 protein or other relevant biomaterials to any of the following respects: (a1) synthesis of the breast milk oligosaccharide; and (a2) synthesis of lactose-N-tri-saccharide II and/or lactose-N-neotetraose, wherein HaHex74 protein is from Haloferula sp., and an amino acid sequence is shown as SEQ ID No.1. The beta-N-acetylhexosaminidase is high in enzyme production level, excellent in hydrolysis characteristics and highin glucoside transformation activity, and has important application value in synthesis of the breast milk oligosaccharide by an enzyme method.

A clean an effective preparation of oligoxylose is carried out by: making xylan solution bydissolving xylan in water or buffering liquid, adding xylanase into it to obtain an enzyme hydrolystic liquid, adding alcohol, centrifuging to obtain supernatant, separating to obtain oligoxylose. It costs low, has no pollution, and is effective. Its product likes natural one.

The invention discloses recombinant escherichia coli, a method for preparing phospholipase C and application. The recombinant escherichia coli BL21(DE-3)-pET-28a-L.m-phospholipase C (PLC) of which the target gene is from listeria monocyogenes (L.monocytogenes) is utilized as a fermentation strain to carry out liquid fermentation, so as to prepare phospholipase C. The method disclosed by the invention is simple in enzyme production process, short in production cycle and low in cost. A glycerophosphate bond on glyceryl phosphatide C3 can be specifically hydrolyzed to produce diglyceride (DAG) by using recombinant phospholipase C; plant crude oil (soybean oil, colleseed oil, rice bran oil and the like) is degummed by phospholipase C.

The invention discloses a fibrinolytic enzyme-producing Bacillus subtilis and a fermentation method and application thereof. The Bacillus subtilis is isolated and sieved from a traditional Chongqing fermented food, i.e., fermented soya beans. The Bacillus subtilis DC27 is collected in China Center for Type Culture Collection under CCTCC NO: M 2015573. The invention further provides a liquid fermentation method of the Bacillus subtilis, and the fibrinolytic enzyme in obtained fermentation broth has an activity of up to 4506 IU/ml. The Bacillus subtilis is from foods and can secrete the high-activity fibrinolytic enzyme; the Bacillus subtilis is safe and nontoxic, and is applicable to the development of functional foods or drugs for preventing or relieving thrombotic diseases; and the Bacillus subtilis has a promising application prospect.

A method for preparing scutellarin by enzymatic hydrolysis skullcap glycoside glucose aldehydic acid is disclosed. Its advantages include no pollution, high percent conversion and high enzyme output.