7 results about "Cholestene" patented technology

The present application relates to a kind of high 4-HBC mycobacterium genetic engineering bacteria and its construction method, and its application in the microbial fermentation preparation 22-hydroxy-23, 24-bis-norcholesta-4-en-3-ketone of 4-HBC.The high 4-HBC mycobacterium genetic engineering bacteria is constructed as follows: with mycobacterium as chassis, kshA1, kshA2, kstD1, kstD2, kstD3 gene is knocked out in turn, hsd4A gene is knocked out, ltp3, ltp4 gene is overexpressed, and the high 4-HBC mycobacterium genetic engineering bacteria is obtained.The genetic engineering bacteria provided in the present application can produce 4-HBC, greatly improve the production efficiency of steroidal drugs, help to improve the conversion rate of substrate, reduce production cost, and reaction condition is mild, environment-friendly, suitable for vigorous popularization and application, with higher economic benefit and social benefit.

A method of treating a cancer (e.g., a colorectal cancer, a liver cancer, a breast cancer, or a lung cancer) in a subject in need thereof by administering to the subject a therapeutically effective amount of (i) a bacterium that converts intestinal cholesterol to 4-cholesten-3-one (4-C-3) or (ii) 4-C-3. Also disclosed is a recombinant Escherichia coli TOP10 bacterium comprising a nucleic acid encoding cholesterol oxidase (CHOX), which is capable of converting cholesterol to 4-C-3 and can be used to treat gastrointestinal tract cancers including a colorectal cancer or lung cancer.

The invention provides a method for synthesizing 4-cholestene-3-ketone based on microorganisms of a cholesterol oxidase truncation. The invention specifically provides a method for producing 4-cholestene-3-ketone, which comprises the following steps: culturing a host cell to obtain a culture product; wherein the host cell can synthesize cholesterol and heterologously express cholesterol oxidase; the cholesterol oxidase truncation is selected from cholesterol oxidase without a complete signal peptide function; and extracting and separating the target product 4-cholestene-3-ketone from the culture product of the host cell. According to the invention, the conversion from cholesterol to 4-cholestene-3-ketone can be directly completed in a living cell environment.

The invention provides an enzymatic preparation method of 3 [beta]-hydroxy-23-bisnorcholest-5-ene-22-acid methyl ester, which comprises the following steps: S1, preparing 3-oxo-23, 24-dinorcholest-4-ene-21-carboxylic acid methyl ester into an intermediate ester compound, and S2, preparing 3 [beta]-hydroxy-23-bisnorcholest-5-ene-22-acid methyl ester into 3 [beta]-hydroxy-23-bisnorcholest-5-ene-22-acid methyl ester; s2, hydrolase and ketoreductase are added into the intermediate ester obtained in the step S1, enzyme catalysis is carried out, then purification is carried out, and the 3 beta-hydroxy-23-bisnorcholest-5-ene-22-acid methyl ester is obtained through preparation. According to the enzymatic preparation method provided by the invention, a cheap and easily available phytosterol fermentation product is taken as a raw material, 3 #-site esterification is carried out through a chemical method to obtain an intermediate ester, and then a target product is finally obtained with the yield of up to 64% through one-step catalytic reaction of hydrolase and ketoreductase. The process has no loss except conversion, and the intermediate is relatively stable, so that the reaction product is relatively high in yield and good in quality, and is suitable for large-scale industrial production.

The invention discloses a blood metabolism marker for evaluating the effect of interfering and improving lipid metabolism by intestinal strains and application of the blood metabolism marker, and belongs to the technical field of biomarkers. The blood metabolism marker comprises 13 kinds of common difference metabolites and 6 kinds of target cholic acid metabolites, the 13 kinds of common difference metabolites are cholest-4-ene-3-ketone, 1-phenyl-1H-pyrrole-2, 5-diketone and the like, and the 6 kinds of target cholic acid metabolites are cholic acid, 7-ketodeoxycholic acid, chenodeoxycholic acid and the like. The blood metabolism marker can accurately reflect the improvement effect of intestinal strain intervention on lipid metabolism, can be applied to preparation of a detection reagent or kit for evaluating the intervention effect of the intestinal strain, and can also be used for evaluating the improvement effect of lipid metabolism and monitoring the intervention effect of the intestinal strain. A detection index with high specificity and high sensitivity is provided for evaluating the effect of interfering the lipid metabolism by the intestinal strain, and the method has an important practical application value.

The invention relates to C7-site hydroxylated monooxygenase for efficiently catalyzing BA and application of the C7-site hydroxylated monooxygenase, and belongs to the technical field of synthetic biology. The amino acid sequence of the C7-site hydroxylated monooxygenase is as shown in SEQ ID NO: 2. Meanwhile, the invention also provides a gene sequence for coding the C7-hydroxylated monooxygenase and a recombinant genetically engineered bacterium for expressing the C7-hydroxylated monooxygenase. Compared with an original enzyme, the structurally modified mutant disclosed by the invention has the advantages that the catalytic ability and the protein stability are remarkably improved, the conversion rate of a substrate BA (22-hydroxy-23, 24-norcholest-4-ene-3-ketone) is increased to 95%, the Tm value of zymoprotein is increased to 66.51 degrees, the stability is remarkably improved, and the mutant has the advantages of being high in yield, good in stability and good in stability. The method has the advantages that the method is simple and convenient to operate, the product 7-OH-DBA (7, 22-dihydroxy-23, 24-norcholest-1, 4-diene-3-ketone) can be efficiently generated, and the efficient utilization of the substrate is realized.

Methods of preventing and / or treating ischemia, organ dysfunction and / or organ failure, including multiple organ dysfunction syndrome (MODS), and necrosis and apoptosis associated with organ dysfunction / failure, are provided. For instance, the methods involve contacting organ(s) with an oxygenated cholesterol sulfate (OCS), e.g. 5-cholesten-3,25-diol, 3-sulfate (25HC3S). The organ(s) may be in vivo (e.g. in a patient that is treated with the OCS) or ex vivo (e.g. an organ that has been harvested from a donor and is to be transplanted).