676 results about "Transaminase" patented technology

The invention belongs to the technical field of biological medicines, and particularly discloses an epidermal growth factor-linker polypeptide-collagen, and the linker polypeptide has an amino acid sequence as shown in SEQ ID No.1. The linker polypeptide capable of connecting the EGF and the collagen is designed and synthesized, the N end of the linker polypeptide simulates an EGF receptor structure and can be combined with the EGF, and the linker polypeptide is connected with the EGF through non-covalent Van der Waals force, so that the structure of the EGF cannot be damaged; the C terminal of the linker polypeptide is rich in lysine residues, and can be covalently crosslinked to collagen through glutamine transaminase to form EGF-linker polypeptide-collagen. The protein solution taking EGF-linker polypeptide-collagen as active ingredients is low in cytotoxicity and high in biological safety, and meanwhile, the action time of the epidermal growth factor can be remarkably prolonged. In addition, a stable repairing microenvironment can be formed on the surface of the endometrium, and the endometrium injury repairing effect is remarkably improved.

The invention discloses a method for preparing an animal adipose tissue simulant by using soybean protein and application of the animal adipose tissue simulant, and belongs to the technical field of deep processing of vegetable protein. Soybean protein is used as a raw material, the soybean protein and polysaccharide are compounded and heated to form thermal induction gel, then microgel particles are obtained through mechanical shearing and crushing, and recombinant soybean protein emulsion gel, namely the animal adipose tissue simulant, is prepared through vegetable oil emulsification and glutamine transaminase covalent cross-linking. The fat-reducing sausage is used as a substitute of pig back fat and is applied to development of fat-reducing sausages. The animal adipose tissue simulant prepared by the invention has texture and melting characteristics similar to those of animal pig back fat. The invention provides the preparation method of the plant-based fat simulant, which meets the requirements of environmental protection and health and sustainable production, not only meets the requirements of texture and melting characteristics of solid fat, but also provides technical support for high-value application of soybean protein in the field of plant meat foods.

The invention belongs to the technical field of biological catalysis, and particularly relates to four R-transaminase and application thereof in asymmetric synthesis of chiral N-heterocyclic amine active drug intermediates. The amino acid sequences of the R-transaminase disclosed by the invention are shown as SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5 and SEQ ID NO. 7. Four kinds of R-transaminase are obtained through gene mining and screening, and recombinant escherichia coli is constructed and expressed by adopting a gene engineering technology. The obtained R-transaminase can be used for synthesizing chiral drug intermediates (R)-3-amino-N-Boc-pyrrole (1b) and (R)-3-amino-N-Boc-piperidine (2b) with high stereoselectivity, and has a good application prospect.

The invention relates to preparation of high-stability mascarpone cheese and application of the mascarpone cheese in baking, and belongs to the technical field of cheese preparation. The glutamine transaminase is combined with a citric acid-sodium citrate system, so that the excessive denaturation of protein is reduced, and the texture and flavor of the cheese are improved. Trehalose and xanthan gum are added for locking water and resisting freezing, and an antioxidant is combined for use, so that the freeze-thaw stability and oxidation resistance of the cheese are improved, the cheese is protected from texture change and oxidative damage in the freezing and unfreezing processes, and excellent texture and flavor can still be kept after multiple times of circulation. Arabic gum is modified through octenyl succinic anhydride, amphipathy is obtained, whey separation is prevented, whey loss is reduced, and the cheese taste is improved.

The invention relates to an application of Danjie emodin in preparation of drugs for preventing or treating drug-induced liver injury. The liver injury is drug-induced liver injury induced by acetaminophen. According to the invention, the effective component Danleaf emodin in the traditional Chinese medicine rhubarb is used for preventing and treating drug-induced liver injury induced by acetamido, and the traditional Chinese medicine composition has the characteristics of safety, reliability and small toxic and side effects. Starting from the whole, multi-target intervention is performed on drug-induced liver injury induced by acetaminophen, the levels of glutamic-pyruvic transaminase, glutamic oxalacetic transaminase and malondialdehyde in plasma are remarkably reduced, the obvious anti-inflammatory and liver cell apoptosis inhibiting effects are achieved, the good clinical application prospect is achieved, and a new way is provided for prevention and treatment of liver injury.

The invention provides a branched chain amino acid transaminase gene and application thereof, and belongs to the field of gene engineering. According to the invention, the branched chain amino acid transaminase gene HvBCAT5 is found for the first time, and the gene has obvious positive correlation with the accumulation of valine (Val), isoleucine (Ile) and leucine (Leu) of crops. The invention provides an important target gene HvBCAT5 for crop quality improvement, and has important value for increasing the content of branched chain amino acid of crops and improving dietary nutrition. The HvBCAT5 gene as well as a recombinant vector, a recombinant bacterium and a transgenic plant thereof have good application prospects in the fields of food and feed industry.

The invention discloses an L-aspartic acid transaminase mutant and application thereof. The L-aspartic acid transaminase mutant is obtained by performing single mutation on the 104th amino acid of the amino acid sequence of L-aspartic acid transaminase. The L-aspartic acid transaminase mutant is used for synthesizing an L-phenylglycine derivative. The L-aspartic acid transaminase mutant is from escherichia coli, homologous sequence analysis and molecular docking simulation are carried out, site-specific mutagenesis verification is combined, and single-point mutation is carried out on the 104 amino acid to obtain the T104S mutant. The L-aspartic acid transaminase mutant disclosed by the invention can be used for efficiently catalyzing to generate an L-phenylglycine derivative, and compared with a wild type, the specific activity of the T104S mutant to o-chlorophenoxyacetic acid is 3.2 times that of the wild type.

The present invention provides a phycocyanin-collagen composite hydrogel and its preparation method and application. The present invention utilizes transglutaminase to catalyze the combination of phycocyanin and collagen to form a new, more stable collagen; transglutaminase acts between protein molecules, utilizes lysine and glutamic acid in phycocyanin and collagen to transfer acyl groups, and cross-links to form new chemical bonds, thereby enhancing the binding strength of phycocyanin and collagen; the hydrogel prepared by the present invention can be used to prepare medical materials and combined with skin injury sites to solve the problem that traditional dressings are easy to adhere to wounds and easily penetrated by tissue fluid and become infected. In addition to ensuring basic air permeability, it reduces the adhesion of the dressing, increases moisture retention and antioxidant properties, and avoids secondary damage when replacing a new dressing. The hydrogel prepared by the present invention has a certain antioxidant capacity and can be used to prepare products with antioxidant effects.

An artificial synthesis method for malonyl-CoA and use thereof are provided. By means of heterologous expression of an aminotransferase and a malonyl-CoA reductase, an artificial synthesis pathway for synthesizing malonyl-CoA by using β-alanine (β-ala) as a precursor is constructed as follows: firstly, under catalysis of a transaminase, β-ala transfers amino groups to α-ketonic acid (such as pyruvic acid, oxaloacetic acid, or α-ketoglutaric acid, etc.), to form an intermediate product 3-oxopropanoate and a corresponding amino acid; the 3-oxopropanoate generates malonyl-CoA under the action of the malonyl-CoA reductase. This pathway addresses the defects of the natural malonyl-CoA synthesis pathway, such as low carbon utilization, consumption of energy substance ATP, release of greenhouse gas CO2, and strict regulation of pathway enzymes, a pyruvate dehydrogenase (PDH) and an acetyl-CoA carboxylase (ACC), thereby achieving high yielding of products using malonyl-CoA as a precursor, including flaviolin, octanoic acid, phloroglucinol, pentadecaheptaene, natamycin, and spinosad.

The invention provides construction and application of a hyaluronic acid recombinant bacterium, and belongs to the fields of biochemical engineering, synthetic biology, genetic engineering and metabolic engineering, the recombinant corynebacterium glutamicum contains a hyaluronic acid synthase gene, a UDP-glucose dehydrogenase gene and a glutamide-6-phosphate fructose transaminase gene; the hyaluronic acid synthase gene, the UDP-glucose dehydrogenase gene and the glutamine-6-phosphate fructose transaminase gene are respectively connected with a sequence RBS1, a sequence RBS2 and a sequence RBS3; the recombinant bacterium provided by the invention also contains a dCas9 protein gene dCas9 and any one of genes sgRNA1, sgRNA2, sgRNA3, sgRNA4, sgRNA5 and sgRNA6, and the recombinant bacterium provided by the invention is high in hyaluronic acid yield and low in production cost.

The invention belongs to the technical field of biological medicines, and particularly discloses an insulin-like growth factor-1-linker polypeptide-collagen, and the linker polypeptide has an amino acid sequence as shown in SEQ ID No.1. The N end of the designed and synthesized linker polypeptide simulates an IGF-1 receptor structure and can be combined with IGF-1, the C end of the linker polypeptide is rich in lysine residues and can be covalently cross-linked to collagen through glutamine transaminase, the attachment time of the IGF-1 in endometrium is prolonged, and endometrium thickening and endometrium injury repair are effectively promoted. The protein solution for promoting endometrial injury repair provided by the invention can form a stable repair microenvironment on the surface of the endometrial, promote the number recovery of endometrial glands with intrauterine adhesion, reduce the fibrosis ratio and promote the thickening of thin endometrial, has the advantages of high histocompatibility, simplicity in preparation, obvious improvement effect and the like, and can be used for preparing a medicine for treating endometrial injury. The method has no potential toxic influence on cells and has a good clinical application prospect.

The art does not provide systematic and reproducible methods to personalize dosing of multiple immunosuppressive medications after transplantation. This invention provides a method to systematize multi-drug immuno suppression management in tissue and organ transplantation using an artificial intelligence-based complex systems approach. In embodiments of this invention, immunosuppression drug dose, blood drug concentrations, donor-derived fraction of cell free DNA (dd-cfDNA %), and aspartate aminotransferase are used to indicate allograft status or a proxy for allograft status, to generate a phenotypic response surface to produce individual treatment modalities and dosages using empirically determined unique coefficients. This surface is used to calculate appropriate immunosuppression drug doses associated with the desired outcome for that patient. Embodiments of this disclosure are directed to identifying optimized combinations of inputs for the complex system of the immunosuppressed transplant patient in order to avoid transplant rejection while avoiding unnecessary toxicity and maintaining a robust enough immune response to fight infection.

The invention belongs to the technical field of bioengineering, and relates to a transaminase mutant, a recombinant genetically engineered bacterium and application of the transaminase mutant in catalytic synthesis of (R)-1-Boc-3-aminopiperidine.The transaminase mutant is obtained by conducting single-point or combined mutation on the 131 site and / or the 197 site of an amino acid sequence shown in SEQ ID NO.2; the mutation sites comprise that the 131 phenylalanine is mutated into aspartic acid, threonine or tyrosine, and / or the 197 lysine is mutated into arginine or leucine. Experimental results show that compared with wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants, especially single-point mutants MyTA1-F131Y and MyTA1-K197R and a combined mutant MyTA1-F131Y-K197R, are all remarkably improved, and compared with the wild type transaminase, the catalytic activity, the thermal stability and the organic solvent tolerance of the obtained mutants are all remarkably improved. The mutant MyTA1-F131Y-K197R can be used for efficiently catalyzing asymmetric amination of N-Boc-3-piperidone to synthesize (R)-1-Boc-3-aminopiperidine, the conversion rate of the (R)-1-Boc-3-aminopiperidine after the (R)-1-Boc-3-aminopiperidine reacts for 24 hours under the condition that the substrate concentration is 100 g / L can reach 90% or above, and the mutant MyTA1-F131Y-K197R has a good industrial application prospect.

The invention discloses an omega-transaminase progenitor enzyme mutant for efficient catalytic synthesis of (R)-chiral amine compounds and application, and relates to the technical field of molecular biology. The omega-transaminase mutant is obtained by mutating an ancestor enzyme Ancaa-101 of omega-transaminase from Aspergillus terreus, and the amino acid sequence of the omega-transaminase mutant is as shown in SEQ ID NO. 6. The invention also discloses a preparation method of the omega-transaminase mutant. Compared with an ancestor enzyme, the activity of the omega-transaminase ancestor enzyme mutant on a non-natural substrate, namely 1-acetyl naphthalene, N-tert-butyloxycarboryl-3-piperidone or 3-oxopyrrolidine-1-carboxylic acid tert-butyl ester, is remarkably improved.

The invention relates to the field of bioengineering, in particular to an enzyme composition, a product and application of the enzyme composition in preparation of Ectoine. The invention provides an enzyme composition. The enzyme composition comprises a mutant of 2-hydroxy-4-aminobutyric acid oxidase, a mutant of keto transaminase, a mutant of hydroperoxide hydrolase, a mutant of Ectoine synthetase, a mutant of pyruvic acid-CoA oxidase and lactobacillus helveticus lactic dehydrogenase. According to the method disclosed by the invention, a plurality of mutant enzymes are utilized for combined transformation for the first time, and cheap 2-hydroxy-4-aminobutyric acid is taken as a starting raw material to quickly and efficiently produce Ectoine; the enzyme method has the characteristics of easiness in control, short conversion time, few generated impurities and the like, so that the method has unique advantages in the aspects of low-cost and large-scale mass production of Ectoin.

The invention relates to a transaminase mutant, a coding gene, an engineering bacterium and an application of the transaminase mutant in synthesis of N-Boc-3-aminopiperidine. A mutant library of transaminase is constructed through protein engineering modification, a series of better transaminase mutants are obtained through screening, and compared with wild transaminase, the enzymatic activity of the mutant is obviously improved. On the basis, the optimal mutant Y113F / R126L is further screened, the enzymatic activity of the optimal mutant Y113F / R126L is 3.9 times that of wild transaminase, and the reaction conversion rate can reach 70.14%.

The present disclosure provides engineered transaminase polypeptides for the production of amines, polynucleotides encoding the engineered transaminases, host cells capable of expressing the engineered transaminases, and methods of using the engineered transaminases to prepare compounds useful in the production of active pharmaceutical agents.

The invention belongs to the technical field of biological catalysis, and particularly relates to application of R-transaminase and a mutant thereof in asymmetric synthesis of chiral amine compounds. The amino acid sequence of the R-transaminase is as shown in SEQ ID NO. 1. The amino acid sequences of the R-transaminase mutants are respectively shown as SEQ ID NO: 2 (G66L), 3 (F127A), 4 (F127M), 5 (G66L / F127M), 6 (G66L / F127A), 7 (G66L / F127M / L160I) and 8 (G66L / H67N / F127M / L160I). The recombinant transaminase mutant obtained by the invention has the advantages of high catalytic activity, strict stereoselectivity, mild reaction conditions and the like for various cyclic ketone and N-heterocyclic ketone compounds.

The invention provides an auxiliary system for a rheumatoid arthritis medication scheme. The auxiliary system is used for matching similar historical medication schemes, related medication costs and treatment results used for treating historical rheumatoid arthritis patients and judging whether transaminase abnormity occurs or not based on the treatment conditions of current patients, analyzing each historical medication scheme, and determining whether the transaminase abnormity occurs or not according to the analyzed historical medication schemes and the analyzed transaminase abnormity. And obtaining scores or grades of three dimensions of safety, effectiveness and economy of each historical medication scheme. And based on the requirements or tendencies of the patient on the medication schemes in three dimensions of safety, effectiveness and economy, screening out corresponding candidate medication schemes from the historical medication schemes and recommending the candidate medication schemes. According to the technical scheme, by means of the computer technology, the corresponding medication scheme is automatically recommended based on the medication tendency of the rheumatoid arthritis patient, decision assistance is provided, the time for adapting the medication scheme for the patient is saved, and the satisfaction degree of the rheumatoid arthritis patient to the medication scheme is improved.

The invention discloses a method for catalytic synthesis of malonic acid by using whole cells, and belongs to the field of bioengineering. According to the invention, escherichia coli BL21 (DE3) is taken as a host, an aspartic acid ammonia lyase gene aspA and an aspartic acid-alpha-decarboxylase gene panD, a succinate semialdehyde dehydrogenase gene yneI and a beta-alanine pyruvate transaminase gene, a beta-alanine pyruvate transaminase gene bauA and a malonyl-coenzyme A reductase gene mcr-C are expressed in modules, and recombinant escherichia coli is constructed. The malonic acid is produced by adopting a whole-cell catalysis method, so that the production period is shortened, the production efficiency of the malonic acid is improved, and the yield of the malonic acid can reach 51.0 g / L after the malonic acid is catalyzed for 24 hours.

The present technical solution discloses a method for producing fermented fish sausages and fermented fish sausages that improve the distinctive flavor of the sausage. The present invention adds 5-pyridoxal phosphate, which is a cofactor that catalyzes the transaminase reaction and can catalyze the leucine in the fish protein to produce α-ketoisocaproic acid, thereby increasing the content of 3-methylbutyraldehyde; by adding a composite fermentation agent, Lactococcus lactis subspecies cremoris contains transaminase, which promotes the catabolism of leucine and isoleucine to produce 3-methylbutyraldehyde and 2-methylbutyraldehyde; Lactobacillus sakei contains α-ketoacid decarboxylase, which promotes the catabolism of leucine to produce 3-methylbutyraldehyde. The present technical solution can improve the fermentation efficiency of naturally fermented sausages and regulate the flavor of the fermented sausages, so that the fermented fish sausages have a rich caramelized aroma, fruity flavor, and bacon flavor, and a delicate, smooth, and elastic taste.

The invention discloses application of spermidine in preparation of a medicine for relieving copper exposure injury, and relates to the field of biological medicine. The invention provides an application of spermidine in preparation of a medicine for relieving copper exposure injury, and the injury is liver injury and / or intestinal injury caused by copper exposure. Spermidine can reduce the copper content in serum and liver, promote the expression of liver copper transporters ATP7B and CTR1, copper death proteins FDX1, LIAS and Lip-DLAT, and reduce the oligomerization of HSP70 and DLAT proteins; meanwhile, transaminase activity in serum and liver and intestinal tissue damage are reduced. The medicine provided by the invention can improve intestinal health, improve liver copper metabolism capability and relieve body injury caused by copper exposure, effectively solves the problem that no reliable medicine exists for related injury caused by copper exposure, and is expected to be developed into a medicine or health food for preventing and treating copper exposure injury.

The invention relates to a heparinized gelatin / sodium alginate pancreas islet packaging material with an anti-fibrosis function as well as a preparation method and application of the heparinized gelatin / sodium alginate pancreas islet packaging material. The preparation method comprises the following steps: adding gelatin into normal saline, soaking, performing autoclaved sterilization to obtain a gelatin solution, then adding epsilon-polylysine hydrochloride into the gelatin solution, and finally adding glutamine transaminase for reaction to obtain an epsilon-polylysine grafted gelatin solution; adding heparin into the solution, and reacting to obtain a heparinized gelatin solution; adding sodium alginate and glutamine transaminase, reacting, diluting, preparing hydrogel microspheres by adopting an electrostatic spraying method, and cross-linking to obtain the hydrogel microspheres. According to the invention, heparin molecules are incorporated into a pancreas islet packaging system, so that fibrosis at the periphery of the microspheres can be relieved, the pancreas islet cells are packaged by using the pancreas islet packaging material to prepare the biological artificial pancreas, the pancreas islet cell proliferation and insulin secretion can be promoted, and the pancreas islet packaging material has a good diabetes treatment effect.

Provided herein is a method for the screening of patients with at-risk MASH (metabolic dysfunction-associated steatohepatitis, formerly named nonalcoholic steatohepatitis or NASH). The combination of levels of five circulating markers, i.e. alanine aminotransferase (ALAT), aspartate transaminase (ASAT), platelets, hsa-miR-34a and YKL-40 in a subject, and eventually the age and gender of said subject, allows to accurately identify non-cirrhotic at-risk MASH. The information obtained from the levels of these markers also allows to screen a subject as being eligible to liver biopsy and / or to classify a subject as a receiver or non-receiver of a treatment of MASH and / or to classify a subject as a responder or non-responder of a treatment of MASH.

The invention relates to an emulsifier composition for meat products as well as a preparation method and application of the emulsifier composition. The emulsifier composition disclosed by the invention is prepared from the following components in parts by weight: 5 to 10 parts of sodium caseinate, 0 to 5 parts of sucrose fatty acid ester, 10 to 15 parts of collagen, 5 to 10 parts of glutamine transaminase, 60 to 70 parts of maltodextrin, 0 to 5 parts of sodium pyrophosphate and 0 to 5 parts of trisodium phosphate. The invention relates to a meat product containing the emulsifier composition, wherein the amount of the emulsifier composition accounts for 0.6%-1% of the total weight of the meat product. According to the invention, the obvious compounding effect among the monomer emulsifier, collagen, phosphate and glutamine transaminase is fully utilized, the functional complementation principle of various raw materials is exerted, and the synergistic interaction effect is generated. On the basis of enhancing the taste of the meat product, the oil and water retention property is improved to a greater extent, so that the yield is improved, and the production cost is reduced.

The invention relates to the technical field of microorganisms, and particularly discloses a recombinant microorganism and application thereof in production of hexamethylenediamine. Compared with an original strain, the recombinant microorganism disclosed by the invention expresses a carboxylic acid reductase CAR variant, a CAR activating enzyme sfp and putrescine transaminase patA; the amino acid sequence of the carboxylic acid reductase CAR variant is as shown in SEQ ID NO. 28; preferably, adipic acid transporters DcaK and DcaP, acyl CoA transferases DcaI and DcaJ, an acylated succinate semialdehyde dehydrogenase sucD, and a 4-aminobutyric acid transaminase gabT are also expressed. By utilizing the recombinant microorganism, 6-aminocaproic acid / adipic acid can be efficiently converted into hexamethylenediamine, and a new method is provided for green biosynthesis of hexamethylenediamine.

The invention provides a transaminase mutant and application of the transaminase mutant in chiral amine synthesis. According to the transaminase derived from Mycobacterium sp. Provided by the invention, in a 1 mL reaction system for catalyzing N-Boc-3-piperidone to generate N-Boc-3-aminopiperidine, the 24-hour conversion rate is 95%, and the e.e. Value of the product is greater than 99%. On the basis, the invention further provides a series of mutants of the transaminase, the relative enzyme activity of the mutants reaches 153%-293%, and the catalytic activity of the transaminase is further improved. Wherein the 34-hour conversion rate of the optimal transaminase mutant is increased to 90% or above under the condition of 120g / L substrate concentration in a 10mL reaction system, and the e.e. Value of the product is greater than 99%. The production technology has the advantages of high catalytic activity, easiness in fermentation, environment friendliness and the like, and has good development and application values.

The invention provides a beta-alanine producing strain as well as a construction method and application thereof. According to the bacterial strain, an acetaldehyde dehydrogenase gene adhE, a lactic dehydrogenase gene ldhA, an acetokinase gene ackA, a pyruvate dehydrogenase gene poxB, a DNA binding transcription inhibition factor coding gene lacI, an aspartate kinase thrA, a pantothenic acid synthase gene panC, an alanine synthesis transaminase gene cycA and an alanine synthesis transaminase gene yfbQ are knocked out from an E.coli W3110 genome, and the bacterial strain is obtained. A phosphoenolpyruvate carboxylase gene ppc, a pyridine nucleotide transhydrogenase coding gene pntAB, an aspartic acid transaminase gene aspC, an aspartic acid decarboxylase gene panD derived from pseudomonas aeruginosa and a beta-alanine transporter gene NCgl0580 derived from corynebacterium glutamicum are integrated at the same time, and the bacillus subtilis is used for producing beta-alanine and has the advantage of high fermentation yield.

The invention provides a method for improving fermentation yield of glutamine, which is characterized in that a coding gene aspB of aspartate transaminase in a microorganism with glutamine production capacity is weakened by utilizing a genetic engineering means to obtain a modified microorganism, and the modified microorganism is used for fermentation production of glutamine. The aspB gene of corynebacterium glutamicum is modified, so that the glutamine generation capacity of corynebacterium glutamicum is higher than that of an unmodified strain.

The invention provides a branched chain amino acid transaminase gene and application thereof, and belongs to the field of gene engineering. According to the invention, the branched chain amino acid transaminase gene HvBCAT3 is found for the first time, and the gene is obviously positively correlated with the accumulation of leucine (Leu) of crops. The invention provides an important target gene HvBCAT3 for crop quality improvement, and has important value for increasing the content of branched chain amino acids of crops and improving dietary nutrition. The HvBCAT3 gene as well as a recombinant vector, recombinant bacteria and transgenic plants thereof have good application prospects in the fields of food and feed industry.