78results about How to "Enzyme stability" patented technology

The present invention discloses a high-yield strain of high temperature resistant 1,4-beta-D-xylanase, and a method for producing the high temperature resistant 1,4-beta-D-xylanase through fermentation of the high-yield strain, and provides partial enzymatic properties of the high temperature resistant 1,4-beta-D-xylanase. After the strain identification, the high-yield strain is named Paenibacillus campinasensis G1-1, and is preserved in the China general microbiological culture collection center. The preservation number of the strain is CGMCC No. 5023. According to the present invention, the high temperature resistant 1,4-beta-D-xylanase produced through the fermentation of the high-yield strain is subjected to separation and purification to obtain a single component, wherein the single component has a relative molecular weight of 41.3 KDa, an optimal operation temperature of 60 DEG C and an optimal operation pH of 7.0; the relatively stable enzyme activity of the high temperature resistant 1,4-beta-D-xylanase is remained in the temperature range of 40-70 DEG C and the pH range of 5.0-9.0; the Paenibacillus campinasensis G1-1 provided by the present invention is applicable for a plurality of fields such as paper making, food, feedstuff and the like, and has a broad application prospect.

The invention discloses a preparation method of immobilized beta-glucosidase in the field of enzyme preparations. According to the method, fructose is adopted as a pore-foaming agent, and the immobilized beta-glucosidase is prepared through the tetraethoxysilane hydrolysis reaction sol-gel method. Different immobilized beta-glucosidase samples are prepared by adding fructose water solutions with different concentrations and beta-glucosidase of different dosages. For the obtained immobilized beta-glucosidase, the invalid adsorption in the enzymolysis process of cellulose can be avoided, so that the enzymolysis conversion efficiency of cellulose is obviously improved, the immobilized enzyme has the advantages of good mechanical strength and multiple times of recycling, and the hydrolysis and saccharifying costs of cellulose are reduced.

The invention provides an enzyme-producing microorganism for hydrolyzing D,L-pantoyl lactone and application and a sifting method of the enzyme-producing microorganism. The method specifically comprises the steps of using an enzyme-producing microorganism strain to hydrolyze D,L-pantoyl lactone, then acidizing D,L-pantoyl lactone to form a converting solution, and conducting high-performance-liquid chromatography chirality online detection on the formed converting solution to quickly sift out strains which have the catalytic activity and stereoscopic selectivity on hydrolyzing D,L-pantoyl lactone. Through the directed sifting method, it is found that a bacterial strain Fusarium verticillioides CGMCC NO.14552 has an obvious and unique advantage in the aspect of hydrolyzing D,L-pantoyl lactone, the hydrolyzing conversion rate of 1-70% D,L-pantoyl lactone within 5-10 h can reach 35% or above, and the ee value of a target product D-pantoyl lactone is larger than 98%.

The invention discloses a compound microbial active filling material for removing sulphur-containing repugnant substances, as well as preparation and application of the compound microbial active filling material. Ochrobactrum anthropi SL1 and aquamicrobium defluvii SU1 related in the invention are isolated bacteria obtained in a reactor for processing repugnant substances, such as H2S, in a water pollution control laboratory of the ecological environment research centre of the Chinese academy of sciences. The bacterial strains are preserved in the general microbial centre of the Chinese general microbial strain preservation and management committee. The preservation numbers are CGMCC No.7400 and CGMCC No.7399 respectively. The compound microbial active filling material disclosed by the invention has the advantages of being large in specific area, high in porosity, good in air permeability, low in resistance, high in bacterial cell loading capacity and less prone to run away; and because of being loaded with ochrobactrum anthropi SL1 and aquamicrobium defluvii SU1, the compound microbial active filling material is difficult to decay and deform in a bioreactor for purifying waste gas, and therefore, the compound microbial active filling material can be used for a long time and is applied to complex processing conditions.

The invention discloses a liquid fermentation method for producing salt-tolerant cellulase from sea aspergillus niger. The liquid fermentation method comprises the following steps: 1) screening soil in 10-20m depth of coastal area of the East China Sea to obtain a strain of sea aspergillus niger; 2) inoculating the sea aspergillus niger into a PDA (potato dextrose agar) culture medium, culturing at 32-40 DEG C for 60-90 hours to obtain slant spores, and preserving the slant spores at 4 DEG C; 3) inoculating the slant spores on a PDA-1 culture medium, and culturing at 32-40 DEG C for 60-90 hours to obtain activated spores; 4) inoculating the activated spores on a slant of an eggplant bottle, and culturing at 42 DEG C for 74 hours to obtain eggplant bottle slant spores subject to expanding culture; and 5) utilizing sterile artificial sea water to dilute the eggplant bottle slant spores obtained by expanding culture to 108 spores/ml, inoculating on a liquid fermentation enzyme productionculture medium, and fermenting for 5-10 days to obtain the cellulase. The liquid fermentation method has the advantages that the produced enzyme has high activity, stability, good salt tolerance and the like, and the produced cellulase can degrade cellulose under the condition of higher salt content, thereby laying a foundation for follow-up application of hydrolysate.

The invention discloses a feed compound enzyme containing neutral protease and a preparation method thereof, and belongs to the technical field of preparation of an enzyme preparation. The feed compound enzyme containing neutral protease is prepared from concentrated malt enzyme, acid proteinase, neutral protease, acidic xylanase, cellulose, beta-glucanase, pectinase, amylase, seminase, phytase, Chinese herb extracts, a protective agent and an activating agent in a scientific distribution manner. The feed compound enzyme is high in enzyme activity, wide in action condition, strong in stability, and more applicable to feed production. A safe digestive enzyme can be provided for beasts and birds, the digestion burden is relieved, the utilization rate and the growth rate of the raw materials are improved, the environment is protected, joyful natural malt fragrance is brought for the feed, the livestock and poultry appetite is enhanced, meanwhile, the efficacy of the enzyme preparation can be fully played by proper quantity of activating agent under the same condition, the adding amount of the enzyme preparation is saved, the expiration date of the compound enzyme preparation is prolonged by scientific distribution of the Chinese herb extracts, and the immunity of the fed beasts and birds can be improved.

The invention relates to a recombined lipoxygenase PhLOX and a preparation method thereof. The PhLOX has activities of a lipoxygenase, a hydroperoxide lyase and a propadiene oxide synthetase at the same time. The method includes following steps: extracting total RNA from porphyra haitanensis; designing a primer according to a gene sequence of the lipoxygenase; amplifying an overall-length cDNA sequence through an RACE technology; recombining the overall-length cDNA sequence to an expression vector pET-28a; converting escherichia coli E.coli BL21; screening out positive clone; and generating the multifunctional lipoxygenase of a bacteria liquid with an expression quantity being 2mg/L through inducible expression. Compared with a lipoxygenase in the prior art, the recombined lipoxygenase has a plurality of enzyme functions, is free of a strict substrate specifity and allows downstream products in various structures to be formed.

The invention relates to an aspergillus oryzae strain capable of producing beta-fructofuranosidase, as well as a culture method and application thereof. An aspergillus oryzae FS4 strain is collected in China General Microbiological Culture Collection Center (CGMCC) on April 24, 2014, with the collection number of CGMCC No. 9087 and the address of Institute of Microbiology, Chinese Academy of Sciences, Building 3, NO.1 Beichen West Road, Chaoyang District, Beijing. The invention further relates to the culture method and application of the aspergillus oryzae strain. The aspergillus oryzae FS4 strain obtained by screening has the advantages of simple culture, stable genetic nature, high synthesis efficiency and stable enzyme activity, can be applied to the field of synthesis of levan type oligofructoses and the like by a modern bio-engineering technical enzyme method, and further has broad application prospects.

The invention discloses a bacillus thermophilus 701 capable of producing high-temperature and alkali resistant xylanase and belongs to the technical field of microbial fermentation. The bacillus thermophilus is obtained through the steps that a soil sample is collected from a paper mill, a bacillus thermophilus is separated from the soil sample, and ultraviolet mutagenesis and nitrosoguanidine mutagenesis, and screening are performed repeatedly. The xylanase produced by the bacillus thermophilus has the characteristics of strong high-temperature resistance and alkali resistance. The bacillus thermophilus is preserved in the China Center for Type Culture Collection on October 12, 2013, the preservation No. is CCTCC M2013537. The high-temperature and alkali resistant xylanase prepared from the bacillus thermophilus 701 is 350-420 IU/ml in specific activity, 40-75 DEG C in applicable temperature range, 65 DEG C in optimum reaction temperature, completely stable in enzyme activity at the temperature of 75 DEG C, 5.0-11.0 in applicable reaction pH value, completely stable in enzyme activity at the pH value of 11.0, and is 10.0 in optimum reaction pH value.

The present invention provides the field of enhancing proteins and in particular to that of proteins enhanced by molecular change. It provides a variant of a phytase that is termed enhanced in that is has better thermostability and/or activity than the original phytase. The invention also provides a nucleic acid coding for said variant, a cassette or an expression vector containing said variant, a host cell expressing said variant, a composition comprising said variant and uses thereof, principally in the preparation of food additives and animal feed.

The invention discloses a bacterial strain producing high-temperature-resistant beta-galactosidase and a screening method thereof and belongs to the technical field of biological chemistry and protein engineering. The screening method includes the specific and sequential steps of sample collection, primary screening, secondary screening of culture and beta-galactosidase activity assay. Mainly, soil of hot spring land is collected and boiled, then bacterial colonies are screened, and the technical problem how to produce high-enzyme-activity beta-galactosidase at high temperature is solved through a high-temperature culturing method. The bacterial strain is short in beta-galactosidase production period, high in beta-galactosidase production speed, good in genetic stability, thermal stability and pH stability, high in enzyme catalytic activity and high in production safety, and culturing conditions are simple. The bacterial strain has broad prospects in the fields of food industry and medical and biochemical analysis, in particular to the dairy industry.

The invention discloses wheat ration enzyme containing neutral protease and a preparation method thereof and belongs to the technical field of enzyme preparation. The wheat ration enzyme is formed by scientific compounding of bacillus subtilis culture, acid protease, acid xylanase, cellulose, beta-glucanase, amylase, beta-mannase, Chinese herb extract, protective agent and activating agent. Due to the bacillus subtilis culture, the immunity of organism of animals can be improved, the growth of the immune organ is enhanced, the maturity of the structure and the function of the intestine of the animal is enhanced, the daily gain of the animal is increased and the feed conversion ratio is improved. The wheat ration enzyme disclosed by the invention has the advantages that safe digestive enzyme is provided for livestock and poultry, the digestion burden is effectively reduced, the material utilization rate and the growth rate are increased, and the environment is protected; simultaneously, due to use of the proper amount of activating agent, the effect of the enzyme preparation can be fully played under the equal conditions, so that the adding amount of the enzyme preparation is saved; due to the scientific compounding of the Chinese herb extract, not only the quality guarantee period of the compound enzyme preparation is prolonged, but also the immunity of the raised livestock and poultry can be improved.

A process for the production of an optically enriched tertiary alcohol of the formula (2a) or (2b), by reacting an epoxide of the formula (1) with a nucleophilic agent Nu in the presence of halohydrin dehalogenase.

The invention relates to a method for preparing fibrilia by using epicoccum nigrum DB3 strains. The method comprises the following steps: (1) preserving the epicoccum nigrum DB3 strains in a potato dextrose agar medium for cultivation, and adding a freezing buffer solution; (2) inoculating bacteria the amount of which equals to the pick-up amount of an inoculating ring, obtained in the step (1) into the potato dextrose agar medium, and carrying out shaking culture; (3) inoculating the culture medium obtained in the step (2) into a flax lignin fermentation medium, and carrying out shaking culture to obtain a fermentation bacteria liquid; (4) mixing sterilized fiber raw material with the fermentation bacteria liquid obtain in the step (3), carrying out table shaking and removing fermentation liquid to obtain the fibrilia; and (5) mixing the fibrilia with degumming liquid, processing and then rinsing, oiling and drying to obtain the fibrilia. The method in the invention has simple technology, short degumming time and high degumming efficiency, thus being suitable for large-scale industrial production.

The invention relates to a preparation method of a magnetic-cellulose-microsphere-immobilized lipase catalyst. The object is to provide a simple, economic, and efficient preparation method of an immobilized lipase catalyst. The adopted process and method comprise: taking cellulose as a raw material, taking solid calcium carbonate particles as a pore-forming agent, through adoption of a hot melt adhesive conversion method and combination with reverse suspension technology, preparing porous regenerated cellulose microspheres, carrying out epoxidation and amination grafting modification, taking micropores as a reactor for a co-precipitation reaction to prepare magnetic cellulose microspheres, and using a covalent bonding method to immobilize lipase molecules to surfaces and micropores of the magnetic cellulose microspheres. The method is abundant and available in raw materials, and the prepared magnetic cellulose carrier is relatively small in size, uniform in distribution, and large in specific surface area. Compared with a catalyst made from a conventional method, the catalyst is high in enzyme immobilization amount, good in dispersion, and firm in loading. The catalyst is convenient to recycle, and can be widely applied to fields of medicine, environment, and energy.

The present invention relates to stable bioactive substances and preparation thereof. A preparation method includes dissolving a bioactive material in a solution substantially free of solutes to make a bioactive solution, adsorbing the bioactive solution onto dry polysaccharide particles to make bioactive loaded polysaccharide particles, and drying the bioactive loaded polysaccharide particles.

The invention relates to mannanases and a recombinant expression bacterial strain thereof. Expression genes of the mannanases is cloned from penicillium decumbens. Amino acid sequences of the mannanases are shown in the formula of SEQ ID NO: 1. The invention also comprises Pichia pastoris pdman-2 engineering bacteria and Trichoderma reesei pdman-2 engineering bacteria for expression of the mannanases. The novel expression genes of the mannanases are cloned from penicillium decumbens, and the Pichia pastoris pdman-2 engineering bacteria and the Trichoderma reesei pdman-2 engineering bacteria for expression of the novel expression gene are constructed. The mannanase expressed by the Pichia pastoris pdman-2 engineering bacteria has the most suitable pH value of 4.5, the most suitable temperature of 55 DEG C and stable enzyme activity in a pH value of 2.0-7.0, and can resist gastric acid and pepsin. The mannanase expressed by the Trichoderma reesei pdman-2 engineering bacteria has the most suitable pH value of 4.5, the most suitable temperature of 55 DEG C and stable enzyme activity in a pH value of 3.5-8.0, and can resist trypsin. The mannanases have feeding-enzyme application values.

The invention discloses a recombinant vector containing Jerusalem artichoke fructan 1-exohydrolase gene and application thereof in producing alcohol by fermentation. The recombinant plasmid is prepared by the following steps: inserting a Saccharomyces cerevisiae 3-phosphoglyceric kinase gene PGK1 promoter between EcoRV and SpeI enzyme digestion sites of an expression vector pUG6, inserting a CYC1-terminator-fused Jerusalem artichoke fructan 1-exohydrolase gene Ht 1-FEHs between SpeI and HpaI enzyme digestion sites of the expression vector pUG6, and inserting an 18SrDNA sequence segment of Saccharomyces cerevisiae genome between XbaI and BstEII enzyme digestion sites of the expression vector pUG6. The invention also discloses application of the recombinant plasmid in producing alcohol by fermentation. The Jerusalem artichoke inulase genes Ht-FEHI and Ht-FEHII can promote fermentation of the Saccharomyces cerevisiae to produce alcohol, thereby providing a new method for enhancing the alcohol yield of Saccharomyces cerevisiae fermentation.

The invention provides a clothing stain remover and a preparation process thereof. The clothing stain remover is prepared from, by mass, 5-15% of a first surfactant, 5-20% of a second surfactant, 8-15% of 2-methyl-2, 4-pentanediol, 3-10% of polyhydric alcohol, 0.1-0.5% of a chelating agent, 1-3% of organic carboxylates, 1-2% of organic acids, 2-7% of alkylol amine compounds, 2-5% of D-limonene, 0.3-1% of a complex enzyme, 0.05-0.1% of a preservative, 0.02-0.03% of essence and the balance water. According to the technical scheme, the first surfactant and the second surfactant as two nonionic surfactants are combined with 2-methyl-2, 4-pentanediol, polyhydric alcohol, the chelating agent, the organic carboxylates and the organic acids, the combination can make the stain remover have a good stain removing capacity, meanwhile, the enzyme activity of the added complex enzyme can keep stable, and the stain removing capacity is further improved, especially the stain removing capacity of oil stain.