70 results about "Aldo-keto reductase" patented technology

Disclosed are medicaments, pharmaceutical compositions, pharmaceutical kits, and methods based on combinations of a hepatitis C virus (HCV) protease inhibitor and an aldo-keto reductase (AKR) competitor, for concurrent or consecutive administration in treating, preventing, or ameliorating one or more symptoms of HCV, treating disorders associated with HCV, or inhibiting cathepsin activity in a subject.

The invention provides a new aldo-keto reductase mutant and a method using the recombinant aldo-keto reductase mutant or a lyophilized powder thereof to carry out asymmetric reduction. When the new aldo-keto reductase is used to catalyze a substrate with the concentration reaching up to 1000g / L, the optical purity of products still reaches up to 98% or above, and extra addition of an expensive coenzyme is avoided. Compared with other asymmetric reduction preparation methods, a method using the aldo-keto reductase has the advantages of high concentration of the products, high optical purity of the products, mild reaction conditions, environmental protection, simple operation and easy industrial amplification, so the aldo-keto reductase has very good industrial application prospect.

The invention discloses a preparation method of a ricefield eel aldo-keto reductase polyclonal antibody. The preparation method comprises the following steps of: (1) cloning of ricefield eel Eakr gene; (2) establishment of a recombinant expression vector of the ricefield eel Eakr gene; (3) inducible expression and purification of recombinant protein of ricefield eel Eakr; and (4) preparation and Western blot analysis of polyclonal antibody. In the invention, a recombinant expression vector of ricefield eel Eakr gene is established by molecular biology and genetic engineering to induce expression, and purified recombinant expression protein is obtained through affinity chromatography. The ricefield eel Eakr polyclonal antibody prepared by the method disclosed by the invention can meet the experimental requirements of immunohistochemistry and Western blot, realizes in-vitro immunoassay, studies the functions of ricefield eel Eakr as well as ricefield eel immunofluorescent staining laser confocal microscopic observation and also can be applied to the immunodetection of aldo-keto reductase of grass carp, pseudobagrus fulvidraco, megalobrama amblycephala, snakehead and crucian.

The present invention discloses a preparation method of methyl (R)-o-chloromandelate utilizing biocatalytic asymmetric reduction and used recombinant vectors and genetically engineered bacteria. The preparation method comprises the step of carrying out biotransformation reaction on methyl o-chlorobenzoylformate used as the substrate with genetically engineered bacteria wet cells or freeze-dried cells capable of coexpressing recombinant reductase and recombinant glucose dehydrogenase as the catalyst at pH 6-8 in the presence of glucose, wherein the recombinant reductase is a recombinant aldo-keto reductase. The genetically engineered bacteria whole cells can simultaneously express the aldo-keto reductase and glucose dehydrogenase and can achieve high-efficiency regeneration of intracellular coenzyme NADP<+>. By using the preparation method, high-concentrations methyl o-chlorobenzoylformate can be catalyzed and completely transformed into methyl (R)-o-chloromandelate with a single conformation, without adding a coenzyme. Because of expensive coenzyme, the preparation method provided by the invention greatly lowers the production cost, has mild reaction conditions, is environmentally-friendly and simple to operate, and has good industrial application prospects.

The invention discloses an aldehyde ketone reductase KmAKR mutant and application thereof in catalytic synthesis of chiral alcohol. The aldehyde ketone reductase KmAKR mutant is obtained by performing single-site or multi-site mutation on the 30 site, the 302 site, the 109 site, the 196 site, the 23<rd> site or the 213<rd> site of an amino acid sequence shown in SEQ ID NO. 1. The specific enzyme activities of the constructed aldehyde ketone reductase mutants M7 and M9 are increased by 1.1 times and 0.63 times respectively compared with those of an original strain, and the T5015 is increased by 6.3 DEG C and 4.9 DEG C respectively. For the mutant M9, when the feeding amount of a substrate 6-cyano-(5R)-hydroxy-3-carbonyl tert-butyl hexanoate can reach 350g / L, the reaction can be completed within 3.7h, the substrate conversion rate is greater than 99%, the dep value of the product is always kept at 99.5% or above, and the space-time yield reaches 1.82kg / L.d.

The invention discloses an ezetimibe synthesis intermediate and a preparation method and application thereof, the ezetimibe synthesis intermediate has the structure as shown in formula III, and is synthesized by enzyme-chemical method, a chiral hydroxyl compound is produced by asymmetric reduction reaction of a raw material compound under the catalysis effect of aldehyde ketone reductase, and a product can be obtained by cyclization reaction. The ezetimibe synthesis intermediate is used in the preparation of ezetimibe, the process is simple, the concentration of the obtained product is high, and the product has the advantages of high optical purity, mild reaction conditions, environmental friendliness, simple operation, easy industrial amplification, and very good industrial application prospect.

The invention relates to analdo-keto reductase, and application of recombinant expression transformant of the aldo-keto reductaseas a biocatalyst in asymmetric reduction of 2-carbonyl-4-phenylbutanoate to synthesis (R)-2-hydroxyl-4-phenylbutyrate, and belongs to the technical field of bioengineering. The amino acid sequence of the aldo-keto reductase is shown in SEQ ID NO.1. The encodinggene the aldo-keto reductase corresponds to the nucleotide sequence of the amino acid sequence of the SEQ ID NO.1 and is shown in SEQ ID NO.2. The novel aldo-keto reductaseis high in catalytic performance.

The invention relates to a method for preparing fludarabine phosphate by an enzyme method. Fludarabine phosphate is prepared by taking fludarabine as a raw material through an enzyme catalytic reaction in the presence of inorganic salt and coenzyme; an enzyme used in the enzyme catalysis reaction is selected from at least one of phosphotransferase, aldoketoreductase, alcohol oxidase or alcohol dehydrogenase; the pH value of the enzyme catalytic reaction is 7.3-7.6, the reaction temperature is 37 DEG C, and the reaction time is 1-4 hours; the coenzyme is ATP. According to the invention, fludarabine phosphate is prepared by adopting a novel enzymatic process, so that the technical problems existing in some existing chemical processes are solved, and the process is improved compared with the existing enzymatic preparation process; the production cost is reduced; the process is environment-friendly and reaction conditions are mild.