106 results about "Amidinotransferases" patented technology

A D-aminotransferase can be modified so as to efficiently produce (2R, 4R)-monatin having high sweetness intensity from 4-(indol-3-ylmethyl)-4-hydroxy-2-oxoglutaric acid by substituting an amino acid at least at one of positions (positions 100, 180 to 183,243 and 244) involved in efficiently producing the (2R, 4R)-monatin in an amino acid sequence of a wild-type D-aminotransferase represented in SEQ ID NO:2.

The present invention relates to methods and compositions for monitoring, diagnosis, prognosis, and determination of treatment regimens in subjects suffering from or suspected of having a renal injury. In particular, the invention relates to using assays that detect one or more markers selected from the group consisting of Cytoplasmic aspartate aminotransferase, soluble Tumor necrosis factor receptor superfamily member 5, soluble CD40 Ligand, soluble C-X-C Motif chemokine 16, S100-A12, Eotaxin, soluble E-selectin, Fibronectin, Granulocyte colony-stimulating factor, Granulocyte-macrophage colony-stimulating factor, Heparin-binding growth factor 2, soluble Hepatocyte growth factor receptor, Interleukin-1 receptor antagonist, Interleukin-1 beta, Interleukin-10, Interleukin-15, Interleukin-3, Myeloperoxidase, Nidogen-1, soluble Oxidized low-density lipoprotein receptor 1, Pappalysin-1, soluble P-selectin glycoprotein ligand 1, Antileukoproteinase, soluble Kit ligand, Tissue inhibitor of metalloproteinase 1, Tissue inhibitor of metalloproteinase 2, soluble Tumor necrosis factor, soluble Vascular cell adhesion molecule 1, and Vascular endothelial growth factor A as diagnostic and prognostic biomarkers in renal injuries.

The invention discloses aminotransferase, a mutant and application to Sitagliptin preparation. According to the application, wet thalli obtained by performing fermentation culture on recombinant escherichia coli containing aminotransferase encoding genes are used as biocatalysts; Sitagliptin precursor ketone is used as a substrate; dimethyl sulfoxide is used as a latent solvent; phosphopyridoxal is used as a coenzyme; isopropylamine is used as an auxiliary substrate; a trolamine buffer solution with the pH being 8 to 9 is used as a reaction medium; a reaction system is formed; the biocatalytic reaction is performed under the conditions of the temperature being 30 to 45 DEG C and the stirring speed being 100 to 250 r / min; after the reaction is completed, the reaction liquid is separated and purified; the Sitagliptin is obtained. The aminotransferase and the mutant are used as biocatalysts; the latent carbonyl compound of Sitagliptin precursor ketone is directly used as the substrate; meanwhile, biocatalytic reaction is performed by using isopropylamine as the auxiliary substrate and using the pyridoxal phosphate as the coenzyme; the separation and purification is performed; Sitagliptin with high optical purity is prepared. The method has the advantages that the total yield is 76 percent; the product e.e. value reaches 99 percent.

A non-naturally occurring microbial organism has at least one exogenous nucleic acid encoding a MEK pathway enzyme expressed in a sufficient amount to produce MEK. The MEK pathway includes an enzyme selected from an acetoacetyl-CoA dehydrogenase (bifunctional), an acetoacetyl-CoA aldehyde dehydrogenase, a 3-oxobutyraldehyde reductase, a 3-oxobutanol dehydratase, an MEK oxidoreductase, a 3-oxobutyraldehyde aminotransferase, a 4-aminobutan-2-one deaminase, a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP aminotransferase, a 2,4-dioxopentanoate decarboxylase, an AKP deaminase, an acetylacrylate decarboxylase, an AKP decarboxylase, a glutamate dehydrogenase, a 3-oxobutyraldehyde oxidoreductase (aminating) and an AKP oxidoreductase (aminating). A 2-butanol pathway further includes an MEK reductase. A method for producing MEK or 2-butanol includes culturing these organisms under conditions and for a sufficient period of time to produce MEK or 2-butanol.

The invention discloses a method for building high-yield 5-aminolevulinic acid Escherichia coli engineering strains, and belongs to the field of metabolic engineering and micro-biological fermentation. On the basis that key enzyme glutamy tRNA reduction enzyme and glutamyl aminotransferase which are 5-aminolevulinic acid C5 synthesis paths are overexpressed through carriers pACYCDuet-1, uroporphyrinogen III synthase (UROS) hemD codes and coproporphyrinogen III oxidase (CPO) hemF codes from an escherichia coli heme biosynthetic pathway are independently expressed with pCDFDuet-1 or are expressed together, and recombination strains are constructed. By means of fermentation verification, hemD or hemF is expressed separated, hemD and hemF are expressed together, and the ALA yield is obviously improved.

Disclosed is a strain of Escherichia sp., capable of producing O-acetyl homoserine in high yield, with the introduction and enhancement therein of the activity of: homoserine acetyl transferase, aspartokinase and homoserine dehydrogenase; and at least one enzyme selected from a group consisting of phosphoenolpyruvate carboxylase, aspartate aminotransferase and aspartate semi-aldehyde dehydrogenase. Also, a method of producing O-acetyl homoserine using the strain is provided.

The present invention relates to methods and compositions for monitoring, diagnosis, prognosis, and determination of treatment regimens in subjects suffering from or suspected of having a renal injury. In particular, the invention relates to using assays that detect one or more markers selected from the group consisting of Cytoplasmic aspartate aminotransferase, soluble Tumor necrosis factor receptor superfamily member 5, soluble CD40 Ligand, soluble C-X-C Motif chemokine 16, S100-A12, Eotaxin, soluble E-selectin, Fibronectin, Granulocyte colony-stimulating factor, Granulocyte-macrophage colony-stimulating factor, Heparin-binding growth factor 2, soluble Hepatocyte growth factor receptor, Interleukin-1 receptor antagonist, Interleukin-1 beta, Interleukin-10, Interleukin-15, Interleukin-3, Myeloperoxidase, Nidogen-1, soluble Oxidized low-density lipoprotein receptor 1, Pappalysin-1, soluble P-selectin glycoprotein ligand 1, Antileukoproteinase, soluble Kit ligand, Tissue inhibitor of metalloproteinase 1, Tissue inhibitor of metalloproteinase 2, soluble Tumor necrosis factor, soluble Vascular cell adhesion molecule 1, and Vascular endothelial growth factor A as diagnostic and prognostic biomarkers in renal injuries.

Methods and compositions for the stabilization of aminotransferase activity of plasma or serum are provided. Such methods will be useful in the accurate determination of tests associated with liver toxicity.

A non-naturally occurring microbial organism has at least one exogenous nucleic acid encoding a MEK pathway enzyme expressed in a sufficient amount to produce MEK. The MEK pathway includes an enzyme selected from an acetoacetyl-CoA dehydrogenase (bifunctional), an acetoacetyl-CoA aldehyde dehydrogenase, a 3-oxobutyraldehyde reductase, a 3-oxobutanol dehydratase, an MEK oxidoreductase, a 3-oxobutyraldehyde aminotransferase, a 4-aminobutan-2-one deaminase, a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP aminotransferase, a 2,4-dioxopentanoate decarboxylase, an AKP deaminase, an acetylacrylate decarboxylase, an AKP decarboxylase, a glutamate dehydrogenase, a 3-oxobutyraldehyde oxidoreductase (aminating) and an AKP oxidoreductase (aminating). A 2-butanol pathway further includes an MEK reductase. A method for producing MEK or 2-butanol includes culturing these organisms under conditions and for a sufficient period of time to produce MEK or 2-butanol.

The present invention provides a method capable of producing a natural or recombinant protein in high yield.The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses alanine aminotransferase and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide.

The invention provides 1,3-diphenylprop-2-en-1-one derivatives and pharmaceutical compositions comprising the same for treating liver disorders, in particular those requiring the reduction of plasma level of biochemical markers such as amino-transferases. The 1,3-diphenylprop-2-en-1-one derivatives of General Formula (I) have hepatoprotective properties and can be used in methods for treating liver disorders involving the pathological disruption, inflammation, degeneration, and / or proliferation of liver cells, such as liver fibrosis or fatty liver disease.

The invention discloses a transaminase mutant as well as application thereof to preparation of sitagliptin midbody. The application adopts a wet thallus obtained by fermenting and culturing recombinant escherichia coli comprising aminotransferase coded gene as a biological catalyst, adopts sitagliptin midbody precursor ketone as a substrate, adopts dimethyl sulfoxide as a co-solvent and , adopts phosphopyridoxal as co-enzyme, adopts isopropylamine as an auxiliary substrate, adopts a pH 8-9 triethanolamine buffering solution as a reaction medium to form a reaction system to perform the biological catalytic reaction under the conditions that the temperature is 30 to 45 DEG C, and the stirring rate is 100 to 250 r / min, after the reaction is ended, the reaction solution is separated and purified to obtain sitagliptin; and the total yield of the method is about 81 percent, and a e.e. value of the product reaches 99 percent.

The invention discloses a method for weakening hemB gene expression to increase yield of 5-aminolevulinic acid synthesized by escherichia coli and belongs to the field of and microorganism fermentation. On the basis of using carrier pRSFDuet-1 overexpression glutamy-tRNA reductase, pancreatic aldehyde amino transferase, coproporphyrinogen III oxidase and carrier pETDuet-1 overexpression urinary porphyrins III synthase, a promoter of hemB genes is horizontally transformed in a gene group so as to control expression of the hemB genes and surveying influence, on ALA accumulation, of the hemB genes. Shake-flask fermentation verifies that yield of a target product ALA is increased remarkably, and the ALA yield at 30h is 2680mg / L.

The invention discloses recombinant corynebacterium glutamicum for accumulating N-acetylneuraminic acid and application thereof and belongs to the field of genetic engineering. The recombinant corynebacterium glutamicum takes corynebacterium glutamicum as an expression host, and an N-acetylneuraminic acid synthesis way is reinforced through over-expression of a glucosamine-fructose-6 phosphate aminotransferase gene, a glucosamine acetylase encoding gene, a phosphatase encoding gene, an acetylglucosamine isomerase encoding agene and an N-acetylneuraminic acid synthase encoding gene; an N-acetylneuraminic acid transport protein encoding gene in the corynebacterium glutamicum and a related gene in an intracellular N-acetylneuraminic acid decomposition, utilization and metabolism are knocked off to obtain corynebacterium glutamicum genetically engineered bacteria for extracellularly accumulating the N-acetylneuraminic acid and the yield reaches 110mg / L; a foundation is laid for further modifying corynebacterium glutamicum through metabolic engineering to produce the N-acetylneuraminic acid.

The invention provides a medicine, a healthy food and a food additive with high safety and no side effect which are used to resist oxidation and stress, and eliminate a plurality of clinical symptoms due to stress load or body and mind fatigue, and are made from Durio zibethinus Murr shell or extract of Durio zibethinus Murr shell. Particularly, the invention provides the medicine, the healthy food, the food additive and a composition for improving body state, which is available for long-term use with high safety and no side effect; the medicine, the healthy food and the food additive are made from Durio zibethinus Murr shell or extract of Durio zibethinus Murr shell, and aim to improve levels of plasma Alanine amino ALT and MDA under stress load. Additionally, the invention proves that the medicine, the healthy food, the food additives and the composition made by extractions from Durio zibethinus Murr shell or extract of Durio zibethinus Murr shell can improve and relieve a plurality of clinical symptoms caused by oxidation resisting and anti-stress load.

The invention provides 1,3-diphenylprop-2-en-1-one derivatives and pharmaceutical compositions comprising the same for treating liver disorders, in particular those requiring the reduction of plasma level of biochemical markers such as aminotransferases. The 1,3-diphenylprop-2-en-1-one derivatives of General Formula (I) have hepatoprotective properties and can be used in methods for treating liver disorders involving the pathological disruption, inflammation, degeneration, and / or proliferation of liver cells, such as liver fibrosis or fatty liver disease.

The invention relates to a novel aminotransferase, DNA encoding the enzyme, a recombinant vector into which the DNA has been introduced, and a transformant into which the vector has been introduced. Further, the invention also relates to a method for producing an optically active amino compound utilizing the enzyme or transformant. The aminotransferase of the invention has an ability of efficiently converting a ketone compound, particularly a cyclic ketone compound to an optically active amino compound. According to the invention, a method for efficiently producing an optically active amino compound, particularly an optically active cyclic amino compound is provided.

The invention relates to an alanine aminotransferase (ALT) detection kit, belongs to the technical field of clinical in vitro diagnostic reagents, and aims at being capable of providing an alanine aminotransferase detection kit which can effectively eliminate interference of endogenous alpha-ketoglutaric acid and is good in stability to enable an alanine aminotransferase determination result to be more accurate and reliable. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 is prepared from a Tris-HCl buffer solution, L-alanine, a reduced coenzyme (NADH), lactic dehydrogenase (LDH), an enzyme protectant, a stabilizer and a preservative, and the reagent 2 is prepared from a Tris-HCl buffer solution, alpha-ketoglutaric acid, a stabilizer and a preservative. Due to the fact that the enzyme protectant and the stabilizer are added into the ALT detection kit, the stability of the alanine aminotransferase detection kit is improved, and the alanine aminotransferase (ALT) detection kit is suitable for various full-automatic biochemical analyzers, easy to operate, safe in operation and convenient to apply and popularize clinically.

The invention discloses a blood alanine aminotransferase detecting reagent with high stability, and belongs to the technical field of clinical in-vitro detection. According to the blood alanine aminotransferase detecting reagent with high stability, disclosed by the invention, the blood serum alanine aminotransferase (ALT) is detected by using the single reagent method on the basis of an enzyme-coupled continuous detection method, so that the problem that the single reagent in the original formula is unstable is solved, and the reagent is simple and convenient to operate, and facilitates clinical application and popularization.

The invention relates to a preparation method of a chiral aminoheterocyclic compound and a derivative thereof. The preparation method comprises the following step: under the action of an aminotransferase catalyst, a ketone compound, which is as shown in the formula I, reacts with an amino donor to obtain chiral amine as shown in the formula II. The method is simple and feasible. By generating a chiral amino compound from a corresponding ketone compound through one step, the reaction step is greatly shortened, yield reaches 70-90%, optical purity of the product is high, and ee value is stabilized at 96% and above, and the technological condition is stable. In comparison with a traditional chemical method, the method has the following advantages: reaction condition is mild; use of a strong oxidant, a strong reducer and dangerous reagents is avoided; reaction time is short; preparation cost is low; condition is mild; and the process causes little pollution to the environment. The reaction route is as shown in the specification.

The present invention relates to electrochemical detection technology, and is quick detecting method of aspartate aminotransferase vitality. Electrochemical or optical change is produced corresponding to the vitality of the measured matter. The method includes: A. sucking the measured matter liquid drop into the reaction area between electric poles to generate oxidation-reduction on the surface of the electric poles so as to produce electron transfer and form current; and B. converting the aspartic acid into glutamic acid and oxaloacetic acid under the cooperated action of current and auxiliary factors and measuring the amount of glutamic acid and oxaloacetic acid so as to obtain the corresponding aspartate aminotransferase vitality. The present invention has raised response speed and detection sensitivity of current type enzyme sensor and lowered reaction voltage.

The invention discloses a method for increasing heme synthesized from escherichia coli and belongs to the technical field of metabolic engineering. Synthetic route genes of the heme from the escherichia coli are divided into two modules, the two modules are combined at random by the aid of plasmids with different copy numbers, and a heme synthetic route is enhanced on the basis of overexpressing glutamyl-tRNA reductase (a hemA code) and glutamyl aldehyde aminotransferase (hemL code). According to the method, a constructed recombinant escherichia coli engineering bacterium: E.coli DH 5a:Puc19-hemA-hemL+pACYCDuet-2-hemB-hemC-hemD-hemE+pRSFDuet-2-hemF-hemG-hemH is subjected to fermentation in a shaking flask so as to produce heme cells reaching 0.56uM / OD (micrometer / optical density). The method has the advantages that a metabolic engineering strategy is adopted to transform microbiological strains to synthesize the target product, the heme, and certain foundations are established for efficient heme production by microbiological methods.

The invention provides 1,3-diphenylprop-2-en-1-one derivatives and pharmaceutical compositions comprising the same for treating liver disorders, in particular those requiring the reduction of plasma level of biochemical markers such as aminotransferases. The 1,3-diphenylprop-2-en-1-one derivatives of General Formula (I) have hepatoprotective properties and can be used in methods for treating liver disorders involving the pathological disruption, inflammation, degeneration, and / or proliferation of liver cells, such as liver fibrosis or fatty liver disease.

The invention relates to an aspartate amino transferase (AST) detection kit and belongs to the technical field of clinic in vitro diagnostic reagents. The invention aims to provide the aspartate amino transferase detection kit with capability of effectively eliminating endogenous alpha-oxoglutarate interference and high stability in order to achieve more accurate and reliable determination results of aspartate amino transferase. The kit provided by the invention comprises a reagent 1 and a reagent 2, wherein the reagent 1 comprises the following components: Tris-HCl buffer solution, L-asparaginic acid, reduction coenzyme (NADH), malic dehydrogenase (MDH), enzyme protective agent, stabilizer and preservative, and the reagent 2 comprises the following components: Tris-HCl buffer solution, alpha-oxoglutarate, a stabilizer and a preservative. The enzyme protective agent and the stabilizer are added into the AST detection kit provided by the invention so that the stability of the aspartate amino transferase detection kit is promoted; the aspartate amino transferase detection kit is suitable for various full-automatic biochemical analyzers; the operation is simple and safe; the aspartate amino transferase detection kit has convenience in clinical application and popularization.

The invention discloses a method for synthetizing chiral cyclic alkyl amino acid by amino transferase. The commercialized material ketonic acid or corresponding soluble ketonic acid salt compound in the market is selected as an initial material; the initial material is dissolved into phosphate buffer, and added to an amino supply body; pyridoxal phosphate (PLP) and amino transferase main enzyme are added to a system containing the amino supply body and main material ketonic acid or corresponding soluble ketonic acid salt compound to react under constant temperature, and obtaining a product with a high ee value, wherein n is equal to 1, 2, 3, 4, 5, or obtaining a product wherein n is equal to 0 and 1. The method is stable in technological condition, simple to operate, high in yield, low in cost, and suitable for large-scale production, and beneficial for environmental protection; and a novel train of thought and a method are provided for the preparation of chiral cyclic alkyl amino acid compound.

The invention provides 1,3-diphenylprop-2-en-1-one derivatives and pharmaceutical compositions comprising the same for treating liver disorders, in particular those requiring the reduction of plasma level of biochemical markers such as amino-transferases. The 1,3-diphenylprop-2-en-1-one derivatives of General Formula (I) have hepatoprotective properties and can be used in methods for treating liver disorders involving the pathological disruption, inflammation, degeneration, and / or proliferation of liver cells, such as liver fibrosis or fatty liver disease.