44results about How to "Improve enzyme production efficiency" patented technology

The present invention is production process of UV and NTG mutagenizing Bacillus subtilis TQS6 to culture high efficiency expressing chitosanase strain TQK and fermentation producing high activity chitosanase. The chitosanase strain is first slant cultured and then fermentation cultured while introducing oxygen-rich air to increase dissolved oxygen to 30-80 %; and after culturing for 18-24 hr, average enzyme activity may reach 718 mu/ml. High activity chitosanase may be obtained through fermentation in optimized enzyme producing condition. The process of the present invention can obtain chitosanase with high yield, high stability and high activity, and the produced chitosanase is applied in converting chitosan into chitin oligose effectively and economically.

The invention aims to construct recombinant pichia pastoris for heterologous expression of EG II, EG IV and EG V proteins. A method comprises the following specific steps: (1) obtaining of trichodermareesei RNA and CDNA: obtaining mycelia by using a solid induction culture medium, and extracting total RNA and cDNA by using a kit; (2) cloning of a target gene: amplifying a target gene by a reasonable primer PCR; (3) constructing an expression vector: taking Ppic9k as a vector, inserting the target gene into the downstream of a promoter AOX1, wherein the 5 ' and 3 ' end enzyme digestion sites are EcoR I and Not I respectively; (4) obtaining of a recombinant strain: taking bgl I as a linearization site of an expression vector to realize linearization and electrotransformation of the expression vector to obtain recombinant yeast; (5) induction and enzyme production of the recombinant strain: carrying out shaking flask fermentation, and inducing enzyme production by using appropriate methanol; and (6) recombinant protein activity detection: detecting the expression and activity of recombinant protein by SDS-PAGE and Congo red-CMC. The recombinant strain constructed by the method can efficiently express the protein with a his label, and a high-efficiency single enzyme component can be obtained.

The invention relates to a bacterial strain for producing a gel breaking enzyme and an application thereof. The bacterial strain for producing the gel breaking enzyme is the Bacilluslichenformis M1-4 bacterial strain, and the preservation number is CGMCC NO.4511. The M1-4 bacterial strain is used for producing the gel breaking enzyme and preparing a bacteria liquid preparation. The bacterial strain obtained by the invention is high in enzyme production efficiency, and the enzyme output under the industrial fermentation condition is greater than 240 U/ml. The recovery method employed by the invention is relatively high in enzyme yield. The produced fracturing fluid gel breaking enzyme has a high activity within a wide range of temperature and is convenient in operation and less in usage amount. The fracturing fluid gel breaking enzyme has a rapid and even viscosity breaking property which enables the enzyme to completely degrade a guar gum with fewer residues, and the viscosity of a fracturing liquid after gel breaking is less than 5.0 mPa.s. Based on the characteristics of the gel breaking enzyme for cleaning fracturing fluid, the disclosed bacterial strain in the invention can not only be used for the conventional fracturing gel breaker construction operation, but also can be used for a remedial measure of a fracturing operation failure.

The invention relates to an inulase mutant and a preparing method. According to the inulase mutant and the preparing method, directed evolution is carried out on an encoding gene inuI of inulase in vitro with a codon optimization technology, an error-prone PCR technology and a site-directed mutagenesis technology, and the mutant with the inulase activity remarkably improved is screened out. The K<cat> value of the mutant is 3.1 times that of the inulase of an original strain; in addition, the preparing process of the inulase is easy to implement and high in enzyme production efficiency, and the enzyme activity of fermentation liquor in a 30-m<3> system can be 60,000 U/ml. Meanwhile, the invention discloses a DNA sequence, an expression vector and a host cell of the encoding inulase mutant. According to the inulase mutant, the preparing method and the DNA sequence, the expression vector and the host cell of the encoding inulase mutant, high efficiency and simple and convenient operability are achieved.

The invention belongs to the technical field of microorganisms and discloses a compound microorganism culture for preparing carotenoid. The compound microorganism culture comprises bacillus subtilis,rhodopseudomonas capsulate and rhodotorula glutinis. The compound microorganism culture increases yield of carotenoid and lowers culture cost.

The invention provides instant berry powder and a making method thereof and belongs to the field of food processing. The instant berry powder is prepared from the following raw materials in parts by weight: 35-40 parts of blueberry, 25-30 parts of mulberry, 10-15 parts of raspberry, 10-12 parts of strawberry, 6-10 parts of gooseberry, 3-5 parts of ficus carica, 8-12 parts of grape, 3-5 parts of kiwi fruit, 4-6 parts of carambola, 5-8 parts of papaya, 3-5 parts of psidium guajave and 4-8 parts of passiflora edulis. The making method comprises the following steps: freezing the raw materials, pressing, then adding an active substance for enzymatic hydrolysis, filtering an enzymatic hydrolysate and mixing with an auxiliary material and maltodextrin, wherein the auxiliary material is prepared by radix puerariae, poria cocos, radix angelicae, semen nelumbinis and peppermint leaves; drying to obtain the instant berry powder. The instant berry powder provided by the invention is powdery and granular, convenient to eat, better in dissolubility and mouth feeling, specific in berry aroma and sweetness, rich in nutrition and difficult for moisture absorption in air and has the healthcare effects of resisting aging, reducing blood pressure and the like.

The invention relates to a composition for catalyzing degradation of uric acid in intestinal tracts. The composition comprises cordyceps militaris powder or dry enzyme powder, which contains urate oxidase which can be used for catalyzing degradation of uric acid. The invention further provides a preparation method and application of the composition for catalyzing degradation of uric acid in intestinal tracts. According to the composition, uric acid is catalyzed to be degraded into allantoin, the content of uric acid in intestinal tracts can be reduced, the proportion of uric acid, secreted andexcreted to the intestinal tracts, in blood can be increased, the concentration of uric acid in the blood can be reduced, and hyperuricemia and gout diseases can be prevented and treated; and excretion of uric acid in urea can be reduced, and the incidence rate of uric acid calculus can be reduced.

The invention belongs to the technical field of biology and discloses a method for producing carotenoid. The method comprises the following steps: centrifuging a compound microorganism culture; removing supernatant and collecting thalli; then adding acetone and carrying out ultrasonic wall breaking; then standing for 30min; then centrifuging and collecting supernatant and sediment; adding acetonewith the volume the same as that of the sediment into the sediment; carrying out ultrasonic treatment and collecting supernatant; combining the supernatant of the two times; then evaporating and drying at low temperature by utilizing a rotary evaporator; dissolving residues with ethyl ether; then adding a KOH methanol solution and standing for 10 to 30min; then adding a NaCl solution and layeringa solvent; collecting an upper-layer ethyl ether phase and evaporating and drying the ethyl ether phase to obtain a carotenoid product. According to the method disclosed by the invention, the purity of a prepared carotenoid product is relatively high and reaches 94 percent or above, and is converted to the yield which reaches 90 percent or above.

The invention discloses a method used for fermentation enzyme production of Serratia marcescens lipase and improving the enzyme activity of the Serratia marcescens lipase. A derivant is added in a fed batch manner in the fermentation process to greatly improve the output of lipase derived from Serratia marcescens, and a multiple-metal ion combination technology is adopted to activate the lipase without changing the enzyme amount in order to improve the enzyme activity of the lipase, so low-cost and high-enzyme activity is provided for production application. The method promotes application of the lipase derived from Serratia marcescens strains in production, greatly reduces the production cost, and makes fermentation enzyme production and enzyme activity improvement be simple and efficient.

The invention discloses a method for inducing coriolopsis gallica to express laccase by using food waste. According to the method, the food waste is used as a fermentation substrate, provides a carbon source, a nitrogen source and a biological induction factor for the coriolopsis gallica, and promotes mycelial growth and laccase induced expression; and the method mainly comprises the following steps: (1) pretreatment of food processing waste; (2) preparation of a coriolopsis gallica seed solution; (3) preparation of a liquid enzyme production culture medium; and (4) production of the laccase through liquid fermentation. The method for inducing the coriolopsis gallica to express the laccase by using the food waste, which is provided by the invention, is high in enzyme production efficiency, short in fermentation period and low in cost, does not need to additionally add any chemical inducer, meanwhile, is beneficial for promoting the resource utilization of the food waste, and has obvious economic benefits and social benefits.

The invention provides a method for producing neutral phytase through immobilized bacillus fermentation. The method comprises the following steps: (1) preparing a seed solution; (2) carrying out pretreatment of a carrier: modifying a ceramsite carrier with an alkali liquor; (3) preparing immobilized cells: filling the carrier into a fixed bed reactor, and immobilizing thallus cells on the carrierthrough an adsorption effect; (4) carrying out pre-culturing: culturing the immobilized cells for 30-40 hours by using a fermentation culture medium; and (5) carrying out continuous fermentation culture: enabling the fermentation culture medium to flow into a fixed bed reactor at a certain dilution rate for continuous fermentation. The bacillus is immobilized by adopting the alkali-modified carrier, and the method has the advantages of being low in cost, stable in property, non-toxic and good in adsorption effect. According to the method, the immobilized cell technology is combined with the continuous fermentation process, so that the purpose of continuously producing neutral phytase at high intensity is achieved, the fermentation efficiency and the phytase yield are greatly improved, andindustrial production is facilitated.

The invention discloses Sartorya Vuill W-10 with a high cellulase yield. A preservation number of the Sartorya Vuill W-10 is CGMCC No.11991. By fermenting the Sartorya Vuill W-10 disclosed by the invention, high-yield cellulose crude enzyme liquid can be obtained; cellulase filter paper enzyme activity can reach 1.26U/ml; the cellulase obtained from fermentation can be applied to preparing bioethanol, a straw residue enzymolysis rate and an ethanol yield can be effectively improved, and certain application value is achieved. The Sartorya Vuill W-10 disclosed by the invention not only has low production cost, high ethanol yield and favorability for commercial application and implementation of biological ethanol, but also provides a way for comprehensively utilizing biological waste.

The invention relates to the technical field of microorganisms, in particular to bacillus velezensis and an application thereof. The preservation number of the bacillus velezensis Lpl-wx is CGMCC No.17045. The bacterial strain can be applied to preparation of astaxanthin esterases. The bacillus velezensis belongs to prokaryotes, so that the fermentation process is quick, enzyme production efficiency is high, and the astaxanthin esterases can be quickly and efficiently obtained. The bacterial strain can be further applied to preparation of an astaxanthin single body and can effectively degradeastaxanthin esters, the astaxanthin single body can be obtained, a green efficient catalyzing way few in by-products is provided, and a technical basis is provided for preparation and promotion of theastaxanthin single body.

The invention relates to a preparation technology of isomaltulose, and specifically to a preparation technology for producing isomaltulose. The preparation technology comprises the following steps ofpreparing mother liquid, preparing immobilized enzyme-producing bacteria, preparing crude enzyme liquid and isomaltulose mother liquid, and preparing an isomaltulose finish product with the purity being greater 95%; the preparation technology for producing isomaltulose provided by the invention abandons the traditional technology of directly producing isomaltose through microbial fermentation (lowyield), produces sucrose isomerase through microbial immobilized fermentation, enables the sucrose isomerase to perform enzymolysis on sucrose in vitro, wherein the percent conversion reaches 95% orabove, the production cost is reduced, a product completely meets the newest international standards and Chinese national standards, and the production cycle is shortened.

The invention discloses a transglutaminase fermentation culture medium pretreated by acidolysis or enzymolysis and a preparation method and application thereof. According to the method, fish meal components in the culture medium are separately subjected to acid hydrolysis and enzymatic hydrolysis, correspondingly nutrient components in the fish meal components can be utilized by a transglutaminaseproducing strain, the utilization rate of the nutrient components by the transglutaminase producing strain is increased, and the enzyme production efficiency is thus improved. The method has the advantages of simple and convenient operation, high efficiency, low cost and easy application in industrial production.

The invention provides a processing method and application of fish meal in a glutamine transaminase fermentation culture medium, discloses a fish meal hydrolysis method and application of the method in fermentative production of glutamine transaminase, and mainly solves the problems of insufficient fish meal hydrolysis and low fish meal utilization rate of an existing fish meal hydrolysis process.Improvement is performed on the basis of an existing fish meal hydrolysis process, wherein the fish meal is subjected to pulverization, impurity removal, acid hydrolysis, alkali hydrolysis, enzymatichydrolysis and the like so that nutrients in the fish meal can be better utilized, and the fish meal utilization rate is increased. The hydrolysis process is also applied to the fermentative production of the glutamine transaminase, which can significantly improve the utilization rate of the fish meal by glutamine transaminase bacteria, and the produced glutamine transaminase has the advantages of high enzyme activity, high stability and the like. The method is simple, convenient, efficient, low in cost and easy to use in industrial production.

The invention discloses a pichia pastoris engineering bacterium for heterologous expression of a cellulase gene EGV and application of the pichia pastoris engineering bacterium. The engineering bacterium is Pichia pastoris-eg5 and is preserved in China General Microbiological Culture Collection Center (CGMCC) in August 2019, with the preservation number of CGMCC No.18422. An endoglucanase gene (EGV) is obtained from trichoderma reesei through a PCR method, cloned and inserted into a pichia pastoris integrated expression vector pPIC9K, and a pPIC9K-eg5 expression vector is obtained. The vectoris introduced into pichia pastoris GS115 through electrotransformation to obtain recombinant pichia pastoris strains, and the recombinant pichia pastoris strains comprising high-copy integrated plasmids are screened by using antibiotics G418 with different concentrations. The optimal induction condition is determined through primary optimization of shake flask fermentation, and meanwhile, the enzymatic properties of the purified recombinant protease are analyzed.