96 results about "Transaminase" patented technology

The application discloses application of branched-chain amino acid transaminase 1 gene in treatment of maple syrup urine disease, and relates to the field of gene therapy. Maple syrup urine disease caused by BCAT2 gene defect can be effectively treated by up-regulating branched-chain amino acid transaminase 1 (BCAT1), and it is verified for the first time that overexpression of BCAT1 (cytoplasm type / brain type) can systemically save lethal metabolic phenotypes caused by BCAT2 (mitochondrial type / general type) defects. The application breaks the barrier of metabolic division, and proves that expression of non-mainstream metabolic enzyme (BCAT1) in non-classical metabolic organs (liver) is enough to reconstruct BCAA decomposition flux. The application expands the MSUD treatment target spectrum: from “must repair BCKDC” to “can compensate for upstream limiting steps in ectopia”, and provides alternative strategy reserves for BCKD-deficient MSUD.

This invention provides a method for preparing vebergerone intermediate compound 4, comprising a nucleophilic reaction of compounds 2 and 3 to obtain compound 4, wherein R1, R2, R3, and Rc are defined as described in the specification. Regarding substrate preparation, this invention also provides a transaminase mutant and its application method. This invention possesses the technical advantages of inexpensive and readily available raw materials, avoidance of the use of highly toxic and flammable substances, ensuring environmental friendliness in the production process, and high transaminase activity.

The present application relates to a kind of preparation method of antibacterial functional complex with stable structure, impregnation liquid, impregnated paper and veneer, belong to antibacterial functional material and artificial veneer technical field.The core of the present application is to prepare a kind of high-temperature-resistant antibacterial functional complex, its method is as follows: tea polyphenol, liquorice extract and citric acid are compounded, then mixed with gelatin solution, then compounded with chitosan under pH 5.0~6.0 by electrostatic interaction, finally crosslinked by glutamine transaminase catalysis, form "electrostatic-covalent" dual stable network structure.The structure effectively encapsulates natural antibacterial active ingredient, so that it can withstand the high-temperature process of impregnation drying and hot pressing, effectively avoid decomposition.The complex is added as additive in impregnated resin, antibacterial impregnated paper and veneer can be prepared.The product has efficient, long-acting, safe antibacterial function, and the preparation process is compatible with existing impregnated paper production line, easy to realize industrial production.

The present application belongs to the field of leather industry production, and particularly relates to a method for high absorption chrome tanning by using cross-linking enzyme preparation. The method comprises the following steps: water immersion, unhairing and liming, deliming, softening, enzyme-assisted tanning, and acid immersion tanning. The enzyme-assisted tanning is assisted by using cross-linking enzyme preparation. The cross-linking enzyme preparation is not used in combination with the hydrolytic enzyme in the softening step. The cross-linking enzyme comprises glutamine transaminase (TG enzyme). The comprehensive activity of the cross-linking enzyme is controlled to be 1-30 U / ml in the application process, and the activity of the TG enzyme is controlled to be 1-20 U / ml. The process in the present application meets the environmental protection requirements. Firstly, the tanning agent absorption rate in the tanning process is successfully improved, and the waste discharge amount in the tanning process is reduced. Secondly, the cleaner cross-linking enzyme preparation is successfully used to replace the chemical auxiliary agent to improve the tanning agent absorption rate. Thirdly, the process operation is simplified.

The application discloses a kind of high-yield O-succinyl-L-homoserine recombinant escherichia coli and its construction method and application, belong to genetic engineering and fermentation engineering technical field.The recombinant escherichia coli is expressed by expressing anti-feedback inhibition homoserine O-succinyltransferase metA fbr , aspartate ammonia-lyase aspA, aspartate transaminase aspC, 2 ketoglutarate decarboxylase sucA, dihydrothioctic acid succinyltransferase sucB and efflux protein yjeH, knock out metJ, improve intracellular NADPH availability, to enhance OSH synthesis pathway metabolic flow, precursor succinyl coenzyme A supply and product efflux.Using 5 L bioreactor for fed-batch fermentation, after fermentation 60 h, OSH production is as high as 137.4 g / L, and sugar acid conversion rate reaches 50.5%.The application provides engineering strain and technical realization path for the efficient fermentation of OSH.

The present application relates to a kind of targeting bacterial UDP-N-acetylglucosamine enolpyruvate transaminase antibacterial compound and its application, the antibacterial compound is S-(4-chlorobenzyl) chloro isothiourea, its chemical structure is as shown in formula I: I;Or its pharmaceutically acceptable salt, stereoisomer, solvate, crystal form, isotopically labeled or prodrug.The antibacterial compound of the present application can effectively inhibit the growth of klebsiella pneumoniae, including carbapenem-resistant klebsiella pneumoniae and high virulence klebsiella pneumoniae;It also has good bacteriostatic effect on escherichia coli and pseudomonas aeruginosa;It is expected to be applied to the clinical treatment of infection caused by drug-resistant klebsiella pneumoniae and high virulence klebsiella pneumoniae and other pathogenic bacteria, and has important clinical significance and social benefits.

The application discloses a kind of biological immobilized enzyme based on yeast surface display technology and the application of synthesis 2-phenylethanol, using yeast surface display technology, for the first time with Saccharomyces cerevisiae surface protein a-lectin as anchor protein, the key enzyme (transaminase, decarboxylase, ethanol dehydrogenase) involved in 2-PE producing Ehrlich pathway in Saccharomyces cerevisiae is separately or jointly carried out biological fixation, enhance substrate channel effect and proximity effect, in vitro one-step catalysis L-phenylalanine synthesis 2-PE, provide yield.The method for fixing multiple enzymes of Ehrlich pathway in the application can be used to reconfigure the biosynthesis pathway of other natural products in vitro, to provide feasible scheme for the rational design of cell factory.

The application relates to application of a transaminase mutant in synthesis of a key chiral intermediate D-4,4'-diphenylalanine of sacubitril valsartan sodium salt (LCZ696), and belongs to the technical field of biocatalytic synthesis.The mutant is a quadruple mutant of the amino acid sequence shown in SEQ ID NO:1, the amino acid sequence of the transaminase mutant is shown in SEQ ID NO:3, and the corresponding amino acid sequence is shown in SEQ ID NO:4.Compared with a wild-type transaminase parent, the unit enzyme activity is increased by about 30 times under the condition of ensuring the chiral purity of a product, the transaminase has very high stereoselectivity and conversion rate, the industrial production cost of sacubitril and LCZ696 can be further reduced, and the transaminase has good industrial application value.

This invention discloses a novel use of carvedilol or its salts and compositions, as well as the composition and its preparation method. The β-blocker or pharmaceutical composition provided by this invention has good efficacy in preventing, treating, and / or alleviating drug-induced liver injury, effectively reducing mortality caused by this disease; simultaneously, it can significantly reduce serum transaminase levels, alleviate symptoms such as liver and spleen edema, and has a good inhibitory effect on pro-inflammatory cytokines. Furthermore, experimental results further indicate that the combination of the β-blocker and pharmaceutically acceptable excipients (steviosides and / or glycyrrhizate) has good safety and is easily dissolved, released, and / or absorbed, improving the bioavailability of carvedilol and exhibiting a synergistic effect, showing promising application prospects.

The application provides a pyroglutamic acid transaminase mutant and application thereof, the mutant is obtained by mutation on the basis of wild-type pyroglutamic acid transaminase, and specific mutation includes at least one of the following sites: (1) the 13th isoleucine is mutated into arginine; (2) the 35th valine is mutated into arginine; (3) the 183rd asparagine is mutated into aspartic acid; and the mutant is suitable for industrial enzymatic production of L-aspartic acid and the process of amino acids taking L-aspartic acid as a precursor; especially, the I13R / V35R / N183D mutant obtained by modification has high positive catalytic activity and strict direction selectivity, and completely loses the reverse reaction activity of decomposing L-aspartic acid; the mutant is overexpressed with phosphoenolpyruvate carboxylase in L-threonine production strain THRS-6, and a high-efficiency L-threonine synthesis metabolic pathway is constructed, and the yield of L-threonine is significantly improved when glucose is used as a substrate.

This application discloses the application of pyridostigmine in the preparation of drugs for the prevention and treatment of non-alcoholic fatty liver disease (NAFLD). Validation was conducted using animal models of NAFLD induced by three different etiologies: methionine-choline deficiency diet, high-fat diet, and leptin gene knockout. Pyridostigmine significantly improved vagal nerve activity in the model animals (e.g., improving high-frequency and baroreflex sensitivity in heart rate variability), effectively corrected serum lipid metabolism disorders (reducing LDL cholesterol), alleviated hepatocyte damage (reducing ALT and AST), and at the histopathological level, reduced hepatic steatosis, decreased inflammatory cell infiltration, and inhibited collagen deposition, thereby achieving the prevention and treatment of NAFLD.

The present application provides a kind of PRMT5 inhibitor, PRMT5 inhibitor includes: recombinant adeno-associated virus vector AAV-shPrmt5 and GSK3326595;Recombinant adeno-associated virus vector AAV-shPrmt5 includes the shRNA of targeting PRMT5, the nucleotide sequence of shRNA is as shown in SEQ ID No:1.The present application also provides the application of the above-mentioned PRMT5 inhibitor in the preparation of drug for improving primary biliary cholangitis, PRMT5 inhibitor is all PRMT5 gene / protein as target point, by inhibiting the expression of PRMT5 gene / protein, reduce liver tissue and spleen tissue Tfh cell and GC B cell, serum autoantibody and transaminase level, can effectively delay the disease progression of primary biliary cholangitis.

The application discloses a kind of high thermal stability glutamine transaminase mutant and its method of expression in streptomyces, belong to enzyme engineering field.The high thermal stability glutamine transaminase mutant provided by the application can efficiently catalyze casein crosslinking at high temperature, realizes the heterologous efficient expression of glutamine transaminase.Rational mutation is carried out to key amino acid site of glutamine transaminase, and the mutant strain with significantly improved thermal stability is obtained.Among them, the optimal mutant strain FRAPD-TGm2A3 has a half-life of up to 537.91 min at 60 DEG C.The mutant can still efficiently catalyze the crosslinking reaction of casein at high temperature of 85 DEG C.The present application not only lays a solid research foundation for the modification of thermal stability of glutamine transaminase, but also provides a generalizable strategy framework for the rational modification of other thermostable enzymes, and has important industrial application value and academic reference significance.

This invention discloses a lipase mutant and its application in the preparation of defatted dried seahorse. The lipase mutant is derived by mutation of the wild-type transaminase with the amino acid sequence shown in SEQ ID NO.1. This invention significantly enhances enzyme activity, making it suitable for the defatting process of dried seahorse. After enzymatic hydrolysis, the dried seahorse retains its intact shape and color. The application of this lipase mutant is expected to achieve a greener, more efficient, and standardized degreasing process for seahorse, improving the quality stability and market competitiveness of seahorse products, and yielding significant economic and social benefits.

The application relates to the technical field of skin care products, and particularly discloses a skin care freeze-dried powder containing collagen polypeptides and a preparation method thereof. The freeze-dried powder comprises active components, synergistic repair polysaccharides, moisturizers, freeze-drying protectants and preservatives and the like. The core of the freeze-dried powder is that the active components are modified fish collagen peptides which are cross-linked by glutamine transaminase and modified by methyl pyrrolidone silanol ketone, so that the stability and biological activity of the product are significantly improved. Meanwhile, a specific compounded polysaccharide system of beta-glucan, tremella polysaccharide and sodium hyaluronate is adopted in the formula, and a moisturizer prepared by ultrasonic cross-linking of chondroitin sulfate and sodium alginate is adopted, so that the components synergistically act to jointly enhance the repair and moisturizing effects and use skin feeling of the product. The application also provides a corresponding preparation method. Through an optimized freeze-drying process, the problems of traditional freeze-dried powder, such as easy moisture absorption, single effect and poor reconstitution are effectively solved, and finally a skin care freeze-dried powder product with excellent comprehensive performance is obtained.

The application belongs to the technical field of biology and pharmaceutical chemistry, and discloses a transaminase mutant, a method and application of the transaminase mutant in preparation of a key chiral intermediate of rotigotine, and a preparation method of the key intermediate of rotigotine is a biological conversion method with the transaminase mutant as a main enzyme, an amino acid sequence of the transaminase mutant is an amino acid sequence mutated from an amino acid sequence of a wild-type transaminase shown in SEQ ID NO. 1, the amino acid sequence of the transaminase mutant has a mutation site in the mutated amino acid sequence, and has more than 90% homology with the mutated amino acid sequence. When 5-methoxy-2-naphtalone (a compound of formula III) is catalyzed to prepare a key chiral intermediate (S)-5-methoxy-1,2,3,4-tetrahydronaphthalen-2-amine (a compound of formula II) of rotigotine through an enzymatic reaction method, the chiral purity of the product is greater than 99.9%, and the yield of the biological catalysis step is greater than 95%.

A selenium-enriched biotin scalp care essence spray and its preparation method belong to the field of scalp care technology. The spray components are: selenium yeast extract, pearl active peptides, hair follicle repair core enzyme complex, biotin, plant extract micropowder, and the remainder being excipients and deionized water, with a pH of 5.0-5.5. The selenium yeast extract contains ≥60% selenocysteine; the pearl active peptides have a molecular weight <1kDa and an amino acid content ≥90mg / mL; the hair follicle repair core enzyme complex contains ≥2000U / mg superoxide dismutase and ≥50U / mg transglutaminase; and the plant extract micropowder is extracted using supercritical CO2. The use of ultrasonic high-pressure homogenization, iontophoresis encapsulation, and nitrogen-filled aluminum bottle packaging technology solves the problems of low transdermal penetration rate of active ingredients (traditional formulations <8%, this invention increases it to 28%) and poor stability.

This invention relates to the use of a class of deaminases or transaminases and their cofactors or metabolic substrates as drugs for the treatment of polyglutamine diseases. Specifically, this invention discovers that a series of transaminases and deaminases (such as asparagine synthase ASNS) can degrade polyglutamine aggregates by deaminating polyglutamine (polyQ) proteins, thereby treating polyglutamine diseases.

This invention provides a multi-enzyme cascade catalytic reaction system and its application, belonging to the field of enzyme catalysis production technology. Using glycerol as a raw material, it includes a first module, a second module, and a third module. The first module involves the production of glyceric acid from glycerol under the catalysis of alditol oxidase. The second module contains glyceric acid, ATP, and NAD+. + L-glutamic acid is synthesized into phosphoserine via glycerate kinase, D-3-phosphoglycerate dehydrogenase, and phosphoserine transaminase. The third module involves phosphoserine and nucleophiles being catalyzed by phosphoserine thioylase to synthesize L-cysteine ​​or a non-natural L-α-amino acid. The multi-enzyme cascade catalytic reaction system provided by this invention can catalyze a variety of nucleophiles, yields high products, is low in cost, causes minimal environmental pollution, and is easily industrialized.

Compositions and methods for the production of L-glufosinate are provided. The method involves converting racemic glufosinate to the L-glufosinate enantiomer or converting PPO to L-glufosinate in an efficient manner. In particular, the method involves the specific amination of PPO to L-glufosinate, using L-glutamate, racemic glutamate, or another amine source as an amine donor. PPO can be obtained by the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid) or generated via chemical synthesis. PPO is then converted to L-glufosinate using a transaminase in the presence of an amine donor. When the amine donor donates an amine to PPO, L-glufosinate and a reaction by-product are formed. For example, when the amine donor is glutamate (including racemic glutamate or L-glutamate), L-glufosinate and the reaction by-product α-ketoglutarate (KG), also known as oxoglutarate, are produced. Because the PPO remaining represents a yield loss of L-glufosinate, it is desirable to minimize the amount of PPO remaining in the reaction mixture. Degradation, other chemical modification, extraction, sequestration, binding, or other methods to reduce the effective concentration of the by-product, i.e., the corresponding alpha ketoacid or ketone to the chosen amine donor will shift the reaction equilibrium toward L-glufosinate, thereby reducing the amount of PPO and increasing the yield of L-glufosinate.Therefore, the methods described herein involve the conversion or elimination of the alpha ketoacid or ketone by-product to another product to shift the equilibrium towards L-glufosinate.

This invention discloses a vitamin A-functionalized polyphenol-phospholipid complex, its preparation method, and its applications, belonging to the field of pharmaceutical formulation and nanodelivery technology. The phospholipid complex comprises a polyphenol drug, a phospholipid material, and a vitamin A phospholipid ligand. The vitamin A phospholipid ligand is formed by vitamin A or its derivatives and a phospholipid derivative containing amino or other reactive groups. The phospholipid complex forms a phospholipid composite structure through non-covalent interactions between the polyphenol drug and the phospholipid material, and the vitamin A phospholipid ligand is inserted into or assembled into the surface or interior of the phospholipid composite structure. Animal experiments show that this phospholipid complex can reduce serum transaminase levels and alleviate the degree of liver fibrosis, and has good blood compatibility and tissue safety. This invention provides an industrially achievable targeted nano-formulation solution for early intervention in liver fibrosis, with promising application prospects.

The present application relates to the recombinant microorganism for producing 3-amino propanol by fermentation method and its construction method and application. Specifically, the present application provides a construction method of the recombinant microorganism for producing 3-amino propanol, which is introducing the coding gene of omega-transaminase into the microorganism producing 3-hydroxy propionaldehyde, wherein the omega-transaminase can catalyze the conversion of 3-hydroxy propionaldehyde into 3-amino propanol in the presence of amino donor. The present application introduces the omega-transaminase capable of catalyzing the synthesis of 3-amino propanol with 3-hydroxy propionaldehyde as the substrate into the microorganism producing 3-hydroxy propionaldehyde based on gene editing, molecular cloning and other technologies, which first creates a brand new full-path biosynthesis process route of 3-amino propanol.

The application provides a taste-improved linalool embedded microcapsule feed additive and a preparation method thereof. The additive is a microcapsule composed of core material linalool and wall material, and the wall material comprises raw material matrix, black soldier fly larvae powder, fly maggot powder, edible gelatin, edible lactose and emulsifier; the fermented raw material matrix is prepared from bean curd residue, peanut cake, wheat bran and cassava residue through co-fermentation of lactic acid bacteria and yeast. The preparation method comprises the following steps: mixing the raw material matrix and wall material components to prepare a wall material aqueous solution, adding linalool emulsion, and then performing composite coagulation, glutamine transaminase cross-linking solidification and spray drying to obtain the product. In the application, the anti-nutritional factors of raw materials are removed through fermentation, the protein digestion and utilization rate is improved, the pungent smell of linalool is shielded with the help of gelatin and lactose, the volatilization and oxidation of linalool are slowed down, the feed palatability is greatly improved, the product stability is good, and the product can be widely applied to livestock and poultry breeding feed.

This invention relates to the application of PRMT4 inhibitors in the preparation of drugs for treating metabolic-associated steatohepatitis (MASH), belonging to the field of fatty liver disease treatment technology. This invention is the first to discover that PRMT4 inhibitors can serve as a novel drug target for treating MASH. Cellular experiments show that PRMT4 inhibitors inhibit palmitic acid-induced hepatocyte steatosis. Animal experiments confirm that PRMT4 inhibitors can improve hepatic fat deposition, inflammatory response, hepatocyte damage, fibrosis, and insulin resistance in high-fat fed mice, and reduce liver tissue and serum triglyceride and transaminase levels. This invention finds that PRMT4 inhibitors mainly inhibit the activation of YAP, a key transcriptional activator involved in lipid and inflammatory metabolism, and the expression of its downstream target genes. Therefore, PRMT4 inhibitors can be used to prepare drugs for treating MASH, with definite and significant efficacy, and have promising medical application prospects.

The application discloses application of Amomum tsao-ko extract in preparation of medicines for preventing and treating chronic alcoholic liver and intestinal injury, and belongs to the technical field of biotechnology. The extract is applied to prevention and / or treatment of alcoholic liver and intestinal injury and liver-intestinal axis disorder caused by long-term drinking, can reduce contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in blood plasma, improve liver and colon tissue pathological injury, repair intestinal mucosal barrier, restore liver tissue mitochondrial function and regulate expression of key factors of liver-intestinal axis; and the Amomum tsao-ko extract can effectively reduce the contents of AST and ALT in blood plasma of alcohol-induced alcoholic liver and intestinal injury model mice at a lower dose, significantly improve liver and intestinal tissue pathological injury, correct liver-intestinal axis function disorder and regulate expression of liver-intestinal axis related factors of liver tissue; the raw material of the application is from a natural product, and is convenient to collect and high in safety.

This invention discloses an ω-transaminase mutant, its preparation method, and its applications, belonging to the field of biocatalytic organic synthesis technology. Compared with the wild-type ω-transaminase, this ω-transaminase mutant has at least two substitution mutations at the following amino acid sites: positions 57, 86, 153, 180, and 259. The amino acid sequence of the wild-type ω-transaminase is shown in SEQ ID NO:1. The synthesis of (S)-6-methylnicotine using this ω-transaminase mutant exhibits advantages such as high yield, good stereoselectivity (ee>99.5%), and high catalytic efficiency, making it suitable for large-scale production. This provides an efficient preparation route for the supply of (S)-6-methylnicotine and its application in related products.

This invention provides an engineered bacterium for producing ectoine, its preparation method, and its application. The engineered bacterium is prepared using the following steps: using Corynebacterium glutamicum as the starting strain, feedback inhibition of aspartate kinase LysC is relieved, the activity of the gene lysE encoding lysine efflux permease is reduced, and the ectoine synthesis gene cluster ectABC is expressed; based on this, the activity of any one or at least two combinations of aspartate semialdehyde dehydrogenase Asd, aspartate transaminase AspB, or 6-phosphogluconate dehydrogenase Gnd is enhanced to further increase the yield of ectoine.

The application belongs to the field of food, and particularly relates to a yeast enzymatic hydrolysate with high emulsification performance, a yeast protein, a yeast protein sodium salt product, and preparation and application thereof. The emulsification activity of the yeast enzymatic hydrolysate is greater than or equal to 25.0 m2 / g, and the emulsification stability is greater than or equal to 77.0%; the emulsification activity of the yeast protein is greater than or equal to 70.0 m2 / g, and the emulsification stability is greater than or equal to 175.0%; the emulsification activity of the yeast protein is greater than or equal to 70.0 m2 / g, and the emulsification stability is greater than or equal to 175.0%. In the application, the yeast is subjected to complex enzymolysis by alkaline protease, flavour protease, glucanase, mannanase and glutamine transaminase, and then subjected to ionization reaction with sodium hydroxide to obtain a yeast protein sodium salt product. The yeast protein sodium salt product has high emulsification activity, good color and flavor, and can be used as a natural emulsifier, thereby solving the problems of limited production of sodium caseinate, unstable supply and high price.