259 results about "Cytochrome" patented technology

The invention discloses a streptomyces-derived cytochrome P450 enzyme mutant and an application of the streptomyces-derived cytochrome P450 enzyme mutant in synthesis of ursodesoxycholic acid. The cytochrome P450 enzyme mutant is obtained by carrying out single mutation or multiple mutation on the 181st site, the 192nd site, the 249th site or the 288th site of an amino acid sequence shown in SEQ ID NO.2. The cytochrome P450 enzyme mutant has high activity and regioselectivity, low-cost lithocholic acid is used as a substrate, efficient cytochrome P450 enzyme is used for hydroxylating the 7beta site of the lithocholic acid, ursodesoxycholic acid is synthesized in one step, the industrial production cost is greatly reduced, and the cytochrome P450 enzyme mutant has good application prospects in industrial production of ursodesoxycholic acid.

The present invention relates to mutant enzymes with enhanced properties and processes for oxidation of organic compound substrates using such enzymes. More specifically, provided herein are mutations in cytochrome P450 enzymes that enhance their peroxygenase activity and / or alter product selectivity compared to wild-type cytochrome P450 enzymes. Uses for such mutant cytochrome P450 enzymes are also provided, such as for biocatalytic oxidation of carbon-hydrogen bonds via a peroxygenase pathway, or for oxidizing a substrate which is an organic compound.

The invention relates to a mutant based on cytochrome P450BM3 and application of the mutant in chloroaniline biodegradation, and belongs to the field of enzymology. The mutant is obtained by mutating threonine at the 268 site of an amino acid sequence shown in SEQ ID NO.1 into aspartic acid or glutamic acid, mutating alanine at the 328 site into phenylalanine, and mutating alanine at the 328 site into phenylalanine. According to the present invention, the amino acid at the 87 site is mutated into alanine, and the valine at the 78 site is mutated into alanine, such that a variety of mutants such as T268D / A328F, T268E / A328F, T268D / A328F / F87A, T268E / A328F / F87A, T268D / A328F / F87A / V78A and T268E / A328F / F87A / V78A are obtained; according to the present invention, the degradation rate of the mutant on 4-chloroaniline, 2-chloroaniline, 2, 4-dichloroaniline, 3, 4-dichloroaniline and 3, 5-dichloroaniline is achieved; the method has important significance and application value for promoting rapid degradation of chloroaniline pollutants in a water environment and maintaining ecological balance.

The invention belongs to the field of genetic engineering and synthetic biology, and provides a novel cytochrome P450 enzyme derived from cephalotaxus plants, a nucleotide, a carrier and a host of the cytochrome P450 enzyme, and application of the cytochrome P450 enzyme in preparation of cephalotaxane diterpenoid compounds. The cytochrome P450 enzyme provided by the invention has multiple catalytic functions, can efficiently catalyze oxidation of C-5 site and C-19 site of cephalotaxene, lays a solid foundation for analysis of a cephalotaxane diterpene biosynthetic pathway, and provides a new method for obtaining important active molecules such as cephalotaxus hainanensis lactone and the like by adopting a synthetic biological means.

The invention discloses a chemoautotrophic bacteria detection method based on a single cell Raman spectrum and 13C stable isotope labeling. 13C stable isotope is added into a bacterial culture solution or an environmental sample for enrichment culture, centrifugation is performed, cells are collected and used for Raman spectrum detection, and according to the deviation of two characteristic spectrum peaks, namely, the cytochrome c peak deviates from 747 cm <-1 >, 1112 cm <-1 >, 131 cm <-1 > and 1584 cm <-1 > to 725 cm <-1 >, 111 cm <-1 >, 1300 cm <-1 > and 1536 cm <-1 > under exciting light with the wavelength of 532 nm; the peak of phenylalanine shifts from 1002cm <-1 > to 965cm <-1 > under exciting light with the wavelength of 532nm, and chemoautotrophic bacteria can be identified. According to the detection method, the limitation of pure culture is overcome, the bottleneck that the metagenome cannot characterize the carbon sequestration activity is solved, and the single-cell and non-destructive detection of the chemoautotrophic bacteria in the natural environment is realized.

The invention relates to a group of specific amplification primers for mitochondrial genes of procambarus clarkia and application thereof, and belongs to the technical field of molecular biology, and the mitochondrial genes of the procambarus clarkia comprise cytochrome c oxidase I subunit genes and cytochrome b genes. The specific amplification primer of the CO I gene comprises a CO I gene upstream primer and a CO I gene downstream primer, wherein the base sequences of the CO I gene upstream primer and the CO I gene downstream primer are respectively shown as SEQ NO.1 and SEQ NO.2; the specific amplification primer of the Cytb gene comprises a Cytb gene upstream primer and a Cytb gene downstream primer of which the base sequences are respectively as shown in SEQ NO.3 and SEQ NO.4. The invention further discloses a kit for detecting the Cytb gene. According to the amplification method of the mitochondrial gene of the procambarus clarkia, PCR (Polymerase Chain Reaction) amplification is carried out on a DNA (Deoxyribonucleic Acid) template solution of the procambarus clarkia by adopting amplification primers of a mitochondrial COI gene and a Cytb gene of the procambarus clarkia. The procambarus clarkia mitochondrial gene specific amplification primer has high specificity, a single target gene can be stably amplified, the length of the obtained procambarus clarkia mitochondrial COI gene segment reaches about 1000 bp, and the length of the obtained procambarus clarkia mitochondrial Cytb gene segment reaches about 800 bp.

The invention discloses an application of a tobacco cytochrome protein P450 gene, and particularly relates to an application of the tobacco cytochrome protein P450 gene in regulation and control of tobacco dwarfing, and the tobacco cytochrome protein P450 gene is an NtCYP71AU50 gene and / or an NtCYP78A7 gene; the nucleotide sequence of the NtCYP71AU50 gene is as shown in SEQ ID NO. 1, and the nucleotide sequence of the NtCYP78A7 gene is as shown in SEQ ID NO. 3. According to the invention, an editing vector for knocking out NtCYP71AU50 and NtCYP78A7 genes is constructed by virtue of a CRISPR / Cas9 mediated gene editing technology, NtCYP71AU50 and NtCYP78A7 gene editing plants are obtained after genetic transformation, and the plant height, stem girth, waist leaf length, waist leaf width and leaf area of the two gene editing plants are lower than those in a control group. By regulating and controlling the growth and development process of the tobacco plants, the tobacco plants are promoted to be dwarfed, and the problem of shortage of tobacco dwarfing resources is solved, so that application of the tobacco plants in dwarfing close planting cultivation is promoted, the lodging resistance of the tobacco is enhanced, and possibility is provided for increasing the yield per unit area.

The invention discloses a method for improving the heterologous soluble expression efficiency of cytochrome P450, which is characterized in that plasmids containing coding genes of fungus-derived cytochrome P450 truncated protein and plasmids containing molecular chaperones are jointly transferred into host bacteria to obtain recombinant genetically engineered bacteria for improving the heterologous soluble expression efficiency of cytochrome P450. Through host screening, molecular chaperonin and cytochrome P450 gene modification, the soluble expression efficiency of cytochrome P450 in Escherichia coli is improved, and the problems of poor protein solubility and low expression efficiency when CYP450 is expressed in Escherichia coli at present are solved.

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 invention relates to the technical field of plant molecular breeding, in particular to application of PH8 protein or a coding gene thereof in regulation of plant height, ear length or lodging resistance of plants. The PH8 protein is cytochrome P450 CYP78A15, and the plant is an oryza plant. The PH8 protein comprises an amino acid sequence as shown in SEQ ID NO.1. The application comprises the following steps: reducing the expression level of the PH8 protein in the plant, reducing the plant height and spike length of the plant, and improving the lodging resistance of the plant. The PH8 protein related to the plant height of the oryza plant is obtained through screening, and the plant height and the ear length of the oryza plant can be regulated and controlled by regulating and controlling the expression level of the PH8 protein in the oryza plant. The PH8 protein provided by the invention can be used for cultivating oryza plants with strong lodging resistance, does not influence the yield of the oryza plants, and has important application value.

The application discloses a kind of generation (4S) 4-hydroxyisophorone monooxygenase mutant and preparation method thereof, the monooxygenase is cytochrome P450 BM3 Monooxygenase from bacillus megaterium. The cytochrome P450 BM3 Monooxygenase mutant has high stereoselectivity and high reaction activity to catalyze isophorone hydroxylation, mutant R47L / Y51F / F81W / A191T / N239H / I259V / A276T / A328I / A330S / L353I / I401L / L437LL catalyzes isophorone to obtain (4S) 4-hydroxyisophorone diastereomeric excess rate can reach 95%, yield can reach 96%; the cytochrome P450 BM3 Monooxygenase mutant catalyzes isophorone synthesis (4S) 4-hydroxyisophorone process is simple, environmental protection, and industrialization prospect is good.

The present invention relates to a cytochrome P450 monooxygenase mutant and its application in hydroxylation of methylpyrazine compounds, belonging to the technical field of enzyme engineering. The amino acid sequence of the cytochrome P450 monooxygenase mutant provided by the present invention is shown in SEQ ID NO.2. The cytochrome P450 monooxygenase mutant L76S / A83E provided by the present invention exhibits hydroxylation activity towards methylpyrazine substrates, and the reaction conversion rate is greater than or equal to 95%, achieving a breakthrough in the catalytic activity of cytochrome P450 monooxygenase from non-existent to existent. Therefore, the cytochrome P450 monooxygenase and its gene described in the present invention have good prospects for industrial application development.

The invention provides a computer-implemented method and system for non-invasive and step-by-step prediction of cancer risks and a storage medium. The method comprises the following steps: (1) determining a cancer signal score based on the level of a biomarker in a blood sample of a subject; (2) comparing the cancer signal score with a preset threshold value, and determining a primary screening positive subject; (3) further performing cancer risk determination and cancer species prediction by adopting an NGS method on the basis of the primarily screened positive subjects; wherein the biomarker is selected from at least one of AFP (Alpha Fetal Protein), CA125, CA15-3, CA19-9, CA72-4, CEA (Carcino Embryonic Acid), CYFRA (Cytochrome Factor Receptor Amplification) 21-1, ProGRP (Proso-Growth Radical Polymerase Chain Reaction), SCCA (Sequence

The present disclosure is directed, in part, to compounds, or pharmaceutically acceptable salts thereof, for modulating the activity of Cytochrome BD oxidase, or a mutant thereof. The disclosure also provides pharmaceutically acceptable compositions comprising compounds of the present disclosure and methods of using said compositions in the treatment of various diseases and disorders related to Cytochrome BD oxidase. Formula (I) and (II).

The present disclosure provides a P450 cytochrome enzyme for andrographolide synthesis and its application, belonging to the field of bioengineering. The present disclosure uses Saccharomyces cerevisiae CEN.PK2-1D as a host, and implements knockout of ROX1 and GAL80 genes on the genome, and integrative expression of GGPP synthase encoding gene and CPS diterpene synthase encoding gene at ROX1 site; and implements free expression of ApCPR and CYP71A8 and CYP71D10 both with truncated signal peptides, successfully constructing recombinant S. cerevisiae, and achieving de novo synthesis of 3,15,19-Trihydroxy-8(17),13-ent-labdadiene-16-oic acid. Compared with the blank, a response value of a product peak reaches 1.9*106, and this strategy provides necessary reference for analyzing biosynthetic pathway of andrographolide and using metabolic engineering to synthesize andrographolide and related derivatives thereof.

The invention discloses saccharomyces cerevisiae engineering bacteria for efficiently synthesizing mogroside V on the basis of cytoplasm and lipid droplet double-cell area compartment and a construction method of the saccharomyces cerevisiae engineering bacteria. The construction method comprises the following steps: integrating truncated 3-hydroxy-3-methylglutaryl coenzyme A reductase tHMG1, pentenyl pyrophosphate isomerase IDI1, epoxy squalene cyclization enzyme ERG1, squalene synthetase ERG9, choline kinase ScCK, isopentenyl phosphokinase AtIPK, cucurbitadienol synthetase SgCDS, cyclic epoxy hydrolase SgEPH3, cytochrome P450 enzyme CYP87D18 and cytochrome P450 enzyme reductase AtCPR1 at the site of a saccharomyces cerevisiae genome GAL80; an ABC efflux protein PDR11, a glycosyl transferase UGTMG1, a sucrose synthase Susy and a glycosyl transferase SgUGT94-289-3 are integrated at a site Exg1 of a saccharomyces cerevisiae genome. The yield of mogroside V produced by shake flask fermentation of the saccharomyces cerevisiae engineering strain provided by the invention reaches 96.3 mg / L, so that mogroside V is efficiently synthesized from the beginning in the saccharomyces cerevisiae, and the saccharomyces cerevisiae engineering strain has important practical significance for promoting sustainable development of the mogroside industry.

The invention discloses ganoderic acid mass production and application based on cytochrome P450 enzyme CYP512W6, the cytochrome P450 enzyme CYP512W6 in ganoderma lucidum is converted into a corresponding saccharomyces cerevisiae strain for heterologous expression, and three kinds of ganoderic acid, namely TLTOA, 3 beta-TLTOA and DLDOA, are obtained after fermentation. The invention provides a new implementation mode for heterologous biosynthesis of ganoderic acid in saccharomyces cerevisiae.

The present invention belongs to the technical field of anti-cancer drugs, and particularly relates to a cerium vanadate targeted nanoparticle, a preparation method thereof and an application. The cerium vanadate targeted nanoparticle provided by the present invention comprises a drug core, a protective shell layer wrapping the drug core; and a targeting material modified on the outer surface of the protective shell; the drug core comprises CeVO<subgt;4< / subgt> nanoparticles. The cerium vanadate targeted nanoparticle provided by the present invention uses CeVO<subgt;4 nanoparticles as a drug carrier. Since the CeVO<subgt;4 nanoparticles have cytochrome c oxidase-like catalytic activity and cooperate with endogenously overexpressed cytochrome c in cancer cells to reduce and consume O<subgt;2, the oxygen concentration in the tumor microenvironment is effectively reduced. The cerium vanadate targeted nanoparticle provided by the present invention enhances the targeting of the nanoparticle to cancer cells, increases the probability of the nanoparticle entering cancer cells, can activate the drug to play a role in a hypoxic environment, and effectively kills tumor cells.

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.

The present invention discloses a gene mutation site of field mite resistance to abamectin and its application, belonging to the technical field of field resistance detection. The present invention uses the cytochrome P450 detoxifying enzyme CYP392A11 gene as a resistance detection marker for field mites to abamectin, and provides a primer pair for amplifying the CYP392A11 gene to verify whether there is a point mutation, thereby completing the rapid identification of field mite resistance to abamectin. The present invention detects the resistance level of field mites to abamectin through the single-base site mutation in the promoter region of CYP392A11 of field mites, and can quickly detect the resistance level of Tetranychus urticae to abamectin in the field, thereby providing technical support for the rational use of pesticides.

The invention discloses a multiple real-time fluorescent PCR (polymerase chain reaction) primer probe combination, a kit and a detection method for detecting four common animal-derived components in a blood product. The multiplex real-time fluorescent PCR detection system comprises four groups of primer probe combinations of ducks, chickens, pigs and geese aiming at cytochrome b genes, and the sequences of the four groups of primer probe combinations are sequentially shown as SEQ ID No.1-12. According to the present invention, by using the specificity of the target gene, the primer probe combination only performs specific amplification on the target gene segment of the respective species, and is not interfered by other species; the method has the advantages of high throughput, strong specificity, high sensitivity and the like, the detection limit can reach 0.01% (w / w), and the method is suitable for large-scale rapid detection of four common derived components of ducks, chickens, pigs and geese in blood products.

The invention discloses a CYP450 enzyme CYP80Q4 participating in biological synthesis of stephania tetrandra alkaloid and application of the CYP450 enzyme CYP80Q4. According to the technical scheme, the cytochrome P450 enzyme from stephania tetrandra is applied to catalysis of a C-O coupling reaction of a compound to generate bisbenzylisoquinoline alkaloid, the name of the protein is CYP80Q4, and the amino acid sequence is a sequence 2 in a sequence table. After a CYP80Q4 gene is expressed in yeast, a microsome extracting solution containing CYP80Q4 protein is extracted, substrates are screened through a substrate spectrum enzymatic reaction, and 10 groups of obtained substrates can be subjected to a C-O coupling reaction under the catalysis of CYP80Q4 enzyme to generate the bisbenzylisoquinoline alkaloid. The method can be applied to the biosynthesis of the dibenzylisoquinoline alkaloid and / or the effective component tetrandrine of the stephania tetrandra, and the breeding of the stephania tetrandra.

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.

The present invention discloses a dual-channel colorimetric fluorescence composite probe for detecting trypsin, a detection method and an application thereof. The dual-channel colorimetric fluorescence composite probe comprises orange-red carbon quantum dots and cytochrome C with a volume ratio of 1.5:1. The dual-channel colorimetric fluorescence composite probe of the present invention can, on the one hand, conveniently and sensitively detect trypsin in samples including serum and urine samples, which is of great significance for studying the relationship between trypsin and the occurrence and development of diseases; on the other hand, the presence of a trypsin inhibitor can cause changes in the colorimetric fluorescence signal, thereby enabling the preliminary screening of novel trypsin inhibitors, providing a simple and rapid new method for the discovery of novel trypsin inhibitors. This detection method is expected to achieve the rapid diagnosis of trypsin-related diseases and the rapid screening of novel trypsin inhibitors.

The present invention addresses the problem of providing a cytochrome P450 protein by identifying a gene of a Cupressaceae plant involved in the biosynthesis of ferruginol and encoding the cytochrome P450 protein with said gene. The present invention also addresses the problem of providing a new method for producing ferruginol on the basis of said gene.　Provided is a cytochrome P450 protein of an isolated CYP720 family or an active fragment thereof, the protein having an activity of oxidizing the carbon at position 12 of an abietane-type diterpene. The cytochrome P450 protein or an active fragment thereof is derived from a plant selected from the group consisting of Cupressaceae, Taxaceae, Cephalotaxaceae, Podocarpaceae, and Araucariaceae.

The present invention relates to mutant enzymes with enhanced properties and methods of using such enzymes to oxidize organic compound substrates. More specifically, provided herein are mutations in cytochrome P450 enzymes that enhance their peroxygenase activity and / or alter product selectivity as compared to wild-type cytochrome P450 enzymes. Also provided are uses of such mutant cytochrome P450 enzymes, e.g., for biocatalytic oxidation of carbon-hydrogen bonds by a peroxygenase pathway, or for oxidation of organic compound substrates.

This method provides in situ quantification of the overall mitochondrial redox state, the redox state of individual cytochromes such as complex III and IV, and methods for treatment should the redox state be identified as hyperoxidized. Mitochondrial rescue from IRI-mediated injury can be achieved using methylene blue, which acts as an alternate electron donor to bypass complex III.

The invention discloses a metabolic pathway of a clerodane type diterpenoid compound. The metabolic pathway is characterized in that related metabolic genes in the biosynthesis process of annonene, hardwickic acid, hatriwaic acid and divinatorin A are analyzed. The invention further discloses a preparation method of the clerodane type diterpenoid compound, and application of the clerodane type diterpenoid compound in the biosynthesis process of the clerodane type diterpenoid compound in the biosynthesis process of the clerodane type diterpenoid compound. The invention also verifies the function of some cytochrome P450 enzymes in the aspect of producing the clerodane diterpenoid compounds by utilizing the fermentation of yeast engineering bacteria, and lays a foundation for the industrialization of the clerodane diterpenoid compounds and intermediates of the clerodane diterpenoid compounds.

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 invention relates to a method for efficiently synthesizing protoberberine compounds and application of the protoberberine compounds. The invention provides a host cell, and the host cell expresses the following enzymes: an enzyme for catalyzing tyrosine hydroxylation to generate dopa, an enzyme for catalyzing dopamine decarboxylation to generate dopamine, an enzyme for catalyzing dopamine to generate 3, 4-dihydroxyphenylacetaldehyde, and an enzyme for catalyzing dopamine and 3, 4-dihydroxyphenylacetaldehyde to condense to generate norlaudanalkali. The enzyme is used for catalyzing norladanine to generate 3 '-hydroxy combined spiceamine, the enzyme is used for catalyzing the 3'-hydroxy combined spiceamine to generate 3 '-hydroxy nitrogen methyl combined spiceamine, the enzyme is used for catalyzing the 3'-hydroxy nitrogen methyl combined spiceamine to generate reticulatonine, the enzyme is used for catalyzing the reticulatonine to generate golden corydalis alkali, and the enzyme is used for catalyzing the golden corydalis alkali to generate tetrahydrocolumbamine. The enzyme is used for catalyzing tetrahydrocotetrandrine to generate tetrahydroberberine, and the enzyme is used for reducing cytochrome P450 in an oxidation state. According to the invention, efficient synthesis of the protoberberine compound is realized.