72 results about "Cytochrome P450" patented technology

The invention belongs to the technical field of biology, and particularly relates to a construction method and application of a tobacco plant for expressing rare ginsenoside. According to the method, dammarendiol synthase DDS, cytochrome P450 CYP716A47 and glycosyl transferase UGTPg1 / UGTPg45 are added into tobacco, heterologous synthesis of rare ginsenosides CK and Rh2 is achieved, dependence on traditional ginseng cultivation extraction is avoided, and cost is remarkably reduced and efficiency is remarkably improved. The characteristics of short growth cycle and easy culture of tobacco are utilized to overcome the problems of low efficiency and low yield of traditional synthesis; positive transgenic tobacco leaves are dedifferentiated to form callus cells, so that materials are provided for large-scale fermentation, and an industrial production foundation is laid.

The present application relates to a kind of cytochrome P450 oxidase CYP716C52 protein, and the CYP716C52 protein coding gene, the protein can catalyze hydroxylation of maytenic acid C2 position to generate triptolide acid C, in turn participate in triptolide biosynthesis.

This invention relates to a cytochrome P450 oxidase STO1, its encoding gene, and its applications. This invention is the first to clone and identify a cytochrome P450 oxidase gene, STO1, from potato responsible for catalyzing the synthesis of spirogelothoraxone, and to determine the nucleotide sequence and translated amino acid sequence of this gene. This invention provides expression vectors, recombinant microorganisms, or transgenic cell lines containing this gene. This invention constructs an engineered Saccharomyces cerevisiae strain for producing spirogelothoraxone, in which the strong promoter ERG9 is replaced by the weak promoter HXT1 in the strain's genome, and the STO1, AtCPR, tHMG1, and STS genes are integrated. This invention also provides a method for constructing this engineered strain and a method for producing spirogelothoraxone using this engineered strain, which can be used for the preparation of spirogelothoraxone.

Provided are: a method for measuring one or more biomarkers to simply and accurately diagnose diseases associated with changes in the activities of cytochrome P450 metabolic enzymes (hereinafter referred to as "P450s"); and a method for detecting the one or more biomarkers. The methods focus on luminescence-based detection of the enzymatic activities of P450s, and involves reacting one or more P450s present in a sample collected in a low- or non-invasive manner, such as serum or urine, with a precursor of a luminescent / light-emitting substrate, metabolizing a luminescent substrate converted from the precursor to generate luminescence, and measuring the generated luminescence. Since many molecular species of P450s are known, the activities of different molecular species of P450s can be measured for each disease on the basis of luminescence intensity, and the activity pattern of each P450 can be analyzed to detect one or more biomarkers.

Plant cells, plants, seeds and portions of plants modified or genetically engineered to have increased expression or activity of at least one protein selected from the group consisting of trehalose phosphate synthetase (TPS), trehalose phosphate phosphatase (TPP), Protein ALP1-like (At3g55350 or ALPL1), Glycosyltransferase 75D1 (UGT75D1), Cytochrome P450 709B2 (CYP709B2), Cytochrome P450 709B1 (CYP709B1) and Cytochrome P450 72A15 (CYP72A15) are provided herein. Also provided are methods of increasing plant resistance to herbicides and / or abiotic stress by increasing expression of at least one of trehalose phosphate synthetase (TPS), trehalose phosphate phosphatase (TPP), Protein ALP1-like (At3g55350 or ALPL1), Glycosyltransferase 75D1 (UGT75D1), Cytochrome P450 709B2 (CYP709B2), Cytochrome P450 709B1 (CYP709B1) or Cytochrome P450 72A15 (CYP72A15).

The invention provides a cytochrome P450 cholesterol side chain lyase mutant, recombinant saccharomyces cerevisiae for producing progesterone and application of the recombinant saccharomyces cerevisiae. The cytochrome P450 cholesterol side chain lyase mutant is obtained through ESM-2 language model screening, and the catalytic efficiency can be remarkably improved. The efficient recombinant saccharomyces cerevisiae is obtained by constructing the recombinant saccharomyces cerevisiae, optimizing the copy number of key enzymes, introducing an electron transfer system and overexpressing the key enzymes, the yield of progesterone is remarkably increased, and the recombinant saccharomyces cerevisiae is suitable for industrial production of progesterone and has important application value.

The present invention relates to compounds of Formula (I), as well as prodrugs, pharmaceutically acceptable salts, pharmaceutical compositions, methods of use, and methods of preparation thereof. The compounds disclosed herein are useful in the treatment of diseases in which the expression of P-glycoprotein and / or cytochrome P450 (e.g., CYP3A4) is modulated (e.g., cancers that have developed multi-drug resistance).

The application discloses a cytochrome P450BM3 mutant, a carrier, microorganisms and application thereof, and belongs to the technical field of chemical enzyme synthesis.The application discloses a synthetic route of a key drug intermediate dehydro-nandrolone acetate synthesized by a chemical enzyme method, wherein enzyme catalysis and chemical catalysis are included.The enzyme catalysis includes a cytochrome P450 mutant and 17beta-steroid reductase, and a one-pot two-step method is used to catalyze acid to be removed to generate 7beta-hydroxynandrolone.Combined with chemical catalysis, the 7beta-hydroxynandrolone is subjected to dehydration esterification to generate the dehydro-nandrolone acetate.The synthetic route has short steps, high yield and the ability to reduce the production cost of related steroid drugs.

The invention belongs to the technical field of gene detection, and particularly relates to a tetranychus urticae cytochrome P450 gene and application thereof to detection and / or monitoring of avermectin resistance of tetranychus urticae. The CYP392A13 gene shows up-regulated expression in a tetranychus urticae resistant population, and the correlation between the target gene and the avermectin resistance of the tetranychus urticae is further confirmed from the back side and the front side through RNAi and in-vitro metabolic activity tests. Therefore, the specific primer pair, the primer group, the kit and the method for detecting the abamectin resistance of the tetranychus urticae are researched and developed, if the expression quantity of a target gene in a population to be detected is obviously higher than that of a tetranychus urticae sensitive population, the population to be detected has the abamectin resistance; a key molecular tool is provided for establishing an abamectin resistance early warning system, and a drug resistance treatment strategy can be formulated in time.

Described herein are steroid or steroid-like compounds including at least one isonitrile group and which inhibit the activity of a cytochrome P450 enzyme, as well as compositions including the compounds. Also included are methods of inhibiting activity of a cytochrome P450 in a subject having a steroid-responsive cancer, a Mycobacterium tuberculosis infection, a fungal infection, or a trypanosome infection, the method comprising administering to the subject the compound including at least one isonitrile group or a pharmaceutically acceptable salt thereof, in an amount effective to inhibit the CYP activity in the subject.

This invention discloses a herbicide-resistant cytochrome P450 gene, BjCYP71C2, its protein, plasmid, and their applications, belonging to the field of plant genetic engineering technology. The nucleotide sequence of this gene is SEQ ID NO:1 or has at least 80% identity with SEQ ID NO:1. This invention also discloses the protein polypeptide and plasmid encoded by the above gene. The purpose of this gene is to express it transgenic in plants, cultivating herbicide-resistant plants for selective weed control using herbicides.

The application relates to the technical field of molecular markers, and discloses an SNP site associated with turbot albinism, a molecular marker and application, wherein the SNP mutation site is a base T / A mutation at the 50th bp of a coding region of a gene KIT proto-oncogene, receptor tyrosine kinase a (kita); the SNP mutation site is a base A / T mutation at the 137th bp of a coding region of a gene cytochrome P450 3A40-like (LOC118311907); the SNP mutation site is a base G / T mutation at the 371st bp of a coding region of a gene kit ligand a (kitlga); and the SNP mutation site is a base G / A mutation at the 581st bp of a coding region of a gene frizzled class receptor 10 (fzd10). The SNP site associated with turbot albinism, the molecular marker and the application provide a precise screening method for turbot breeding. In the breeding process, the SNP molecular markers are used to determine whether the genotype is albinism or normal, to establish an excellent family with normal body color, to reduce the incidence of abnormal body color, and to improve the overall quality and economic benefits of turbot culture.

The application belongs to the technical field of diagnostic markers, and particularly relates to a pathological diagnosis biomarker combination for adrenal Cushing syndrome and application. The pathological diagnosis biomarker combination comprises low density lipoprotein receptor (LDLR), 3-hydroxy-3-methylglutaryl coenzyme A synthetase 1 (HMGCS1) and cytochrome P450 11B1 (CYP11B1). The area under the ROC curve (AUC) of the pathological diagnosis biomarker combination can reach 0.881 (95% confidence interval (CI): 0.775-987; sensitivity: 85.7%; specificity: 85.0%), which indicates that the combination of LDLR, HMGCS1 and CYP11B1 as the pathological diagnosis biomarker combination for adrenal Cushing syndrome has high accuracy.

The invention belongs to the field of chemical ecology of agricultural pests and pesticide science, and relates to an insecticide composition for improving the sensitivity of agricultural pests to insecticides and application of the insecticide composition. Folic acid is applied to a pest feeding substrate, a host plant or a feed, so that the sensitivity of pests to various insecticides (including neonicotinoids, pyrimidines, pyrethroids, organophosphorus and the like) can be remarkably enhanced. It is found that folic acid can regulate and control a carbon metabolism network in pests and inhibit cytochrome P450 and other detoxification enzyme related pathways, and then the endurance capacity of the pests to insecticides is reduced. The composition and the method are verified in seven important pests such as brown planthopper, sogatella furcifera, spodoptera frugiperda, cotton aphid, chilo suppressalis, panonychus citri and thrips, have the advantages of improving the pesticide effect, reducing the pesticide application amount and delaying the resistance evolution, and can be widely applied to comprehensive treatment of agricultural pests.

The invention discloses a W-5 strain for improving pesticide tolerance of natural enemy insects and a culture method of the W-5 strain, the W-5 strain can well grow in a lowest-salt culture medium with lambda-lambda-cyhalothrin as a unique carbon source, the OD600 of a cultured bacteria solution is continuously increased, and the degradation rate of cyhalothrin pesticide reaches 98.24%. Furthermore, after the tenebrio molitor treated by the W-5 is fed to the hunting fuscipes, the activity of two types of key detoxifying enzymes, namely glutathione S-transferase and cytochrome P450, in the body of the hunting fuscipes can be remarkably improved. After the strain is continuously fed for one week, the W-5 is successfully colonized in intestinal tracts of the hpactor fuscipes, and the survival rate of the hpactor fuscipes under the stress of lambda-lambda-cyhalothrin is increased to 90.625%. Therefore, through double mechanisms of'direct microbial degradation 'and'host detoxification metabolism activation', the W-5 synergistically enhances the resistance of the Harpactor fuscipes to the lambda-lambda-cyhalothrin.

The present disclosure provides a method of preparing an organosilicon compound having at least one silicon-bonded methanol group and / or at least one silanol group from a cytochrome P450 variant that promotes oxidation of a silicon-bonded hydrocarbon group in the presence of an oxidizing agent. The method comprises combining the cytochrome P450 variant, an initial organosilicon compound having at least one silicon-bonded hydrocarbyl group, and a cofactor to obtain a reaction mixture, and exposing the reaction mixture to an oxidizing agent to oxidize the silicon-bonded hydrocarbyl group of the initial organosilicon compound, thereby producing the organosilicon compound having at least one silicon-bonded methanol group and / or at least one silanol group. Also disclosed are cytochrome P450 variants suitable for use in the methods, as well as methods for engineering and optimizing the cytochrome P450 variants. Nucleic acids encoding the cytochrome P450 variants as well as compositions, expression vectors and host cells comprising the nucleic acids are also disclosed.

The application discloses application of a cytochrome P450BM3 mutant in synthesis of allylprogesterone, the cytochrome P450BM3 mutant can catalyze 4, 9 to generate 11 alpha-hydroxy-4, 9, and further synthesis of allylprogesterone is combined with chemical catalysis. That is, the application provides a method for synthesizing allylprogesterone by combining biological enzyme catalysis and chemical catalysis, which not only simplifies a drug synthesis step, improves catalytic selectivity, reduces by-products, improves yield, and has mild reaction conditions, low cost, green environmental protection and high efficiency, and has important application value for promoting the development process of steroid drugs in China.

Provided herein are methods for preparing gamma lactones comprising reacting a carboxylic acid substrate with a heterologous cytochrome P450 (CYP450) protein having carboxylic acid 4-hydroxylase activity.

The present application relates to a cytochrome P450 oxidase, and a gene encoding the cytochrome P450 oxidase, the enzyme is involved in the final key step of the biosynthesis of important active ingredients of Tripterygium wilfordii, triptolide and / or neo-triptolide, and can be used for biosynthesis of triptolide and / or neo-triptolide.

The invention discloses a redox chaperonin PDX mutant for improving steroid hydroxylation catalysis activity of cytochrome P450 and application, according to the PDX mutant provided by the invention, the efficiency of OlePM for catalyzing LCA to generate UDCA is remarkably improved, mutants such as Y34I, Y34S, Y34E, Y34H, R67G, R67D, R67A, R67S, I90L, I90M, I90E, I90V and I90S are excellent in performance, and the UDCA concentration can reach 0.13-0.21 g / L and is improved by 5-8 times compared with that reported in literatures. By improving the electron transfer efficiency of the redox chaperonin, the steroid hydroxylation performance of the cytochrome P450 is remarkably enhanced, a new biocatalyst is provided for efficient and green biosynthesis of UDCA, and the method has important industrial application value.

The embodiment of the invention provides a cytochrome P450 hydroxylase mutant and application thereof. According to the cytochrome P450 hydroxylase mutant provided by the embodiment of the invention, a P450 enzyme single mutant CYP199A2-F185L from Rhodopseudomonas palustris is taken as a template enzyme, and the amino acid sequence of the P450 enzyme single mutant CYP199A2-F185L is shown as SEQ ID NO.1. The encoding gene of the protein is modified by adopting a site-specific mutagenesis technology, so that the encoding gene is subjected to amino acid mutation of F188W + V298L + M360W; the cytochrome P450 hydroxylase mutant provided by the embodiment of the invention can selectively catalyze hydroxylation of cinnamic acid to generate 2, 3-dihydroxycinnamic acid, has excellent biological catalytic activity and selectivity, is simple, convenient and efficient to operate, is suitable for industrial production, and has a good application prospect.

This invention provides a cytochrome P450 gene HvCYP94C1 and its application in improving the resistance of highland barley to powdery mildew, belonging to the field of agricultural biotechnology. The cytochrome P450 gene HvCYP94C1 of this invention can hydroxylate JA and JA-Ile to 12-OH-JA-Ile and 12-COOH-JA-Ile, respectively. Silencing this gene in powdery mildew-sensitive materials can significantly increase the jasmonic acid content, thereby improving the disease resistance of the materials, showing excellent application prospects.

Described herein are compositions and uses thereof comprising (a) methyl 3-((methylsulfonyl)amino)-2-(((4-phenylcyclohexyl)oxy)methyl)piperidine-1-carboxylate or a salt thereof; and (b) an agent for reducing metabolism of (a).

The invention relates to a cytochrome P450 oxidase STO1 as well as a coding gene and application thereof. According to the invention, a cytochrome P450 oxidase gene STO1 in charge of catalytic synthesis of spirovetirone in potatoes is cloned and identified for the first time, and a nucleotide sequence and a translated amino acid sequence of the gene are determined. The invention provides an expression vector, a recombinant microorganism or a transgenic cell line containing the gene. According to the invention, a saccharomyces cerevisiae engineering bacterium for producing spirovetanone is constructed, a strong promoter ERG9 in a genome of the engineering bacterium is replaced by a weak promoter HXT1, and the engineering bacterium is integrated with an STO1 gene, an AtCPR gene, a tHMG1 gene and an STS gene. The invention also provides a construction method of the engineering bacterium and a method for producing spirovetirone by using the engineering bacterium, and the engineering bacterium can be used for preparing spirovetirone.

The invention discloses a cytochrome P450 oxidase HPO and application thereof in synthesis of noottanone, and by combining modification of a target gene of the cytochrome P450 oxidase HPO and metabolic engineering optimization modification of an expression host pichia pastoris genome, the shake flask yield of the noottanone is remarkably increased to 356.93 mg / L; compared with the prior art, the high-yield strain is improved by 185%; furthermore, recombinant pichia pastoris is used for high-density fermentation, so that the yield of noottanone reaches 3365.36 mg / L, and a foundation is laid for industrialization of biosynthesis of noottanone.

The invention belongs to the technical field of gene detection, and particularly relates to a tetranychus urticae cytochrome P450 gene and application thereof to detection and / or monitoring of avermectin resistance of tetranychus urticae. The CYP392A13 gene shows up-regulated expression in a tetranychus urticae resistant population, and the correlation between the target gene and the avermectin resistance of the tetranychus urticae is further confirmed from the back side and the front side through RNAi and in-vitro metabolic activity tests. Therefore, the specific primer pair, the primer group, the kit and the method for detecting the abamectin resistance of the tetranychus urticae are researched and developed, if the expression quantity of a target gene in a population to be detected is obviously higher than that of a tetranychus urticae sensitive population, the population to be detected has the abamectin resistance; a key molecular tool is provided for establishing an abamectin resistance early warning system, and a drug resistance treatment strategy can be formulated in time.

The invention belongs to the technical field of biology, and relates to a cytochrome P450 mutant and application thereof. The mutant is obtained by implementing site-directed mutagenesis of F87A, F81I and A82F on an amino acid sequence shown as SEQ ID NO. 2 in a sequence table. The cytochrome P450 mutant provided by the invention shows high catalytic activity on an unnatural substrate epothilone D, and efficient conversion of the epothilone D is realized. The mutant can catalyze the hydroxylation reaction of epothilone D at a plurality of sites (such as C-21 and C-26) to generate at least four different derivatives, thereby providing a precious compound library for drug screening.

A chemoenzymatic method for synthesizing steroid HE3286 is provided, comprising: screening cytochrome P450 mutant LG-23 capable of catalyzing 7β-hydroxylation of dehydroepiandrosterone; enzymatically converting dehydroepiandrosterone to 7β-hydroxy-dehydroepiandrosterone using the P450 BM3 mutant enzyme; and chemically performing alkynylation at the C17th position carbonyl group to generate steroid HE3286. The steroid HE3286 synthesis method not only features simplified synthetic steps and high catalytic selectivity, but also offers mild reaction conditions, low cost, and environmentally friendly efficiency. This approach holds significant application value for advancing the development of steroid pharmaceuticals.

The present disclosure in various aspects provides host cells and methods for making sesquiterpene components of cypriol oil or oud oil. In some embodiments, the host cells express one or more terpene synthase (TPS) enzymes. In some embodiments, the invention provides host cells producing oxidized sesquiterpene components of cypriol oil or oud oil. In these embodiments, the host cells further expresses one or more cytochrome P450 (CYP) enzymes.

The invention discloses a recombinant saccharomyces cerevisiae strain with high yield of asiatic acid and a construction method of the recombinant saccharomyces cerevisiae strain. The method comprises the following steps: integrating a C-2alpha hydroxylated cytochrome P450 oxidase gene AmCYP716C53 from avicennia marina and a C-23 hydroxylated cytochrome P450 oxidase gene CaCYP716E19 from centella asiatica to a chromosome 911b site of a chassis strain ZY35 by utilizing a CRISPR-Cas9 gene editing technology, so as to obtain ZSA01; then, genes MCH5, FLX1, RIB1, FMN1 and FAD1 are integrated to the 1014a site of the ZSA01, and ZSA02 is obtained; finally, the AmCYP716C53 is integrated to the HMX1 site of the ZSA02, and an engineering strain ZSA03 is constructed. Fermentation results show that ZSA03 reaches 1g / L in a 5L fermentation tank, and the ZSA03 has high-efficiency synthesis and industrial application potential.