14 results about "Cytochrome Reductases" patented technology

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention discloses a heme oxygenase mutant and application of the heme oxygenase mutant in preparation of bilirubin through multienzyme coupling, and belongs to the field of biological catalysis engineering. The forward mutant of the heme oxygenase is obtained through a directed evolution technology, and heme can be efficiently catalyzed to synthesize biliverdin; the bilirubin is synthesized by further introducing biliverdin reductase, coupling cytochrome P450 reductase and formate dehydrogenase, and coupling the two core enzymes with foreign assistant enzymes to construct recombinant escherichia coli to catalyze heme to synthesize bilirubin. In the system, NADPH does not need to be exogenously added, the reaction time is about 4 h, the yield of bilirubin reaches 2.10 g / L, accumulation of intermediate products is not observed in the catalysis process, and the highest yield and the shortest reaction time in bilirubin synthesis reported at home and abroad at present are achieved. Compared with the traditional pig bile extraction, the method disclosed by the invention breaks through the resource limitation, is simple to operate and high in yield, and realizes efficient and green synthesis of bilirubin.

The invention relates to the technical field of biological medicines, in particular to application of panthenol-cytochrome c reductase core protein 2 lactic acid modification as a therapeutic target in preparation of drugs for treating sepsis cardiomyopathy. The invention discloses a key effect of Uqcrc2 K109 site lactic acid modification in SIC and application of the Uqcrc2 K109 site lactic acid modification as a therapeutic target. The Uqcrc2-K109 is found to be a therapeutic target of the SIC through molecular biology methods such as proteomics analysis, lactylation omics analysis and co-immunoprecipitation, and the function of a mitochondrial respiratory chain complex III is recovered in a targeted manner by regulating and controlling the lactylation level of the Uqcrc2-K109, so that a new strategy is provided for treatment of the SIC.

The invention provides a steroid compound 11 alpha-hydroxylase and application of the steroid compound 11 alpha-hydroxylase. The coexpression vector provided by the invention can simultaneously express the steroid compound 11alpha-hydroxylase and cytochrome P450 reductase in escherichia coli or mycobacteria, a genetic engineering strain containing the coexpression vector can be suitable for 11alpha-hydroxyl conversion of various steroid substrates, the specificity is relatively strong, the highest conversion rate can reach 90.39%, and the coexpression vector can be applied to the conversion of 11alpha-hydroxyl of various steroid substrates. And the production technology is simple, efficient and controllable, and has important industrial application value.

The invention belongs to the technical field of gene engineering and microbial fermentation, and particularly discloses a cytochrome P450 reductase coding gene CPR2 from aspergillus nidulans and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO: 1, and the amino acid sequence of the protein coded by the gene is shown as SEQ ID NO: 2. According to the invention, the CPR2 gene (OECPR2) is identified as an optimal electron transfer regulatory element by performing function screening on eight CPR genes annotated in a genome, and the CPR2 gene is over-expressed in an echinocandin B production strain aspergillus nidulans, so that the intracellular electron transfer efficiency is remarkably enhanced, the enzyme catalytic activity of cytochrome P450 is driven, and the cytochrome P450 can be used for preparing an echinocandin B cell. And finally, the yield of echinocandin B is efficiently increased. Experiments show that the engineering strain for over-expression of CPR2 can enable the yield of echinocandin B to reach 1869.97 + / -96.98 mg / L in shake flask fermentation, which is 24.37% higher than that of the original strain. The invention provides a key gene element and an engineering strain for industrial efficient production of echinocandin B, and has important economic value.

The invention belongs to the field of gene engineering, and relates to a gene combination for synthesizing delphinidin, a recombinant plasmid combination, a strain and a method for heterologous biosynthesis of delphinidin. The gene combination comprises the following genes: a flavanone 3-hydroxylase gene TbF3H, a P450 cytochrome oxidase gene TbCYP, a flavanonol 4-reductase TbDFR, an anthocyanin synthetase TbANS and a P450 cytochrome reductase gene LjCPR, and the flavanone 3-hydroxylase gene TbF3H, the P450 cytochrome oxidase gene TbCYP, the flavanonol 4-reductase TbDFR, the anthocyanin synthetase TbANS and the P450 cytochrome reductase gene The invention provides a potential method for replacing traditional artificial cultivation or submerged fermentation acquisition, and lays a foundation for further transforming efficient biosynthesis through metabolic engineering.

The invention belongs to the field of microbial metabolic engineering, and discloses recombinant saccharomyces cerevisiae for synthesizing p-coumaric acid by using acetic acid as well as a preparation method and application of the recombinant saccharomyces cerevisiae. The recombinant saccharomyces cerevisiae has the following characteristics that: tyrosine lyase FjTAL, phenylalanine lyase AtPAL2, cinnamic acid 4-hydroxylase AtC4H, cytochrome P450 reductase AtATR2 and cytochrome B5 CYB5 are subjected to overexpression. The invention also provides a construction method and a domestication method of the recombinant saccharomyces cerevisiae. Compared with recombinant saccharomyces cerevisiae which is not evolved, the recombinant saccharomyces cerevisiae evolved adaptively can well grow in a culture medium which takes acetic acid as a unique carbon source, and p-coumaric acid with higher yield can be synthesized. The invention provides a feasible strategy for synthesizing p-coumaric acid, flavonoid or stilbene natural products by using acetic acid as a carbon source.

This invention discloses a thermophilic recombinant *Yersinia lipolytica* strain producing carotenoids, its construction method, and its applications. The recombinant strain exogenously expresses thermophilic precursor fatty acid desaturase (FAD3) and isocitrate dehydrogenase (IDH2) mutants from *Rhodotorula rubrum*, achieving efficient synthesis at 30°C. Secondly, the metabolic pathway of the recombinant strain is optimized based on a metabolic flux push-pull strategy, overexpressing isopentenyl pyrophosphate isomerase (IDI), farnesyl pyrophosphate synthase (ERG20), phytene dehydrogenase (CarB), and bifunctional phytene synthase (CarRP) to increase precursor throughput. Finally, multiple copies of the genes for astaxanthin synthase (crtS) and cytochrome P450 reductase (crtR) from *Rhodotorula rubrum* are performed to further enhance the carotenoid production capacity of *Yersinia lipolytica*. The thermophilic *Yersinia lipolytica* construction method of this invention is simple, can achieve more efficient carotenoid synthesis at higher temperatures, significantly reduces fermentation costs, and has potential for industrial production.

The application discloses a soluble expression vector of a P450 enzyme for producing indole-3-methylamine and a construction method and application thereof. The construction method is that a nucleotide sequence for coding a P450 enzyme tAMIS protein and a nucleic acid sequence for coding a NADPH-cytochrome reductase CPR protein are expressed in a vector, so that a soluble expression vector of a plant-derived P450 enzyme in Escherichia coli is obtained. The expression vector solves the problem that most plant-derived P450 enzymes cannot exhibit biological activity in the Escherichia coli, and compared with using plants or yeasts as hosts, the time consumed for expressing proteins is reduced. A method for detecting indole-3-methylamine synthesized by the Escherichia coli through an exogenous gene is provided.

The application discloses a kind of heme oxygenase mutant and its multienzyme coupling preparation of application of bilirubin, belong to biological catalysis engineering field.The application obtains the positive mutant of heme oxygenase by directed evolution technology, can efficiently catalyze heme to synthesize chlorophyllin;Further introduce chlorophyllin reductase, coupling cytochrome P450 reductase, formic acid dehydrogenase, and two kinds of core enzyme are coupled with exogenous auxiliary enzyme, constructs recombinant escherichia coli, catalyzes heme to synthesize bilirubin.In the system, without exogenous addition NADPH, reaction time is about 4h, and bilirubin yield is as high as 2.10g / L, and no intermediate accumulation is observed in catalytic process, reaches the highest yield of the synthesis of bilirubin reported at home and abroad, and the shortest reaction time level.Compared with traditional pig bile extraction, the method of the application breaks through the resource limit, and the operation is simple, the yield is high, realizes the efficient, green synthesis of bilirubin.

The invention relates to the field of bioengineering, in particular to an enzyme composition, a strain and application of the strain in preparation of santalol. The present invention provides an enzyme composition comprising: a santalene synthase, a cytochrome P450 monooxygenase, and a cytochrome P450 reductase 2. The invention provides a saccharomyces cerevisiae strain for producing santalol and application of the saccharomyces cerevisiae strain for producing santalol, which is characterized in that a key enzyme santalene synthase variant and cytochrome P450 in a santalol biosynthesis path are integrated onto a saccharomyces cerevisiae genome of a high-yield substrate farnesyl pyrophosphate (FPP), and a path for producing santalol by heterologous metabolism is constructed in saccharomyces cerevisiae; the constructed recombinant saccharomyces cerevisiae strain for producing santalol has an important application prospect in synthesis of santalol.

The present application relates to a kind of CYP11B1-BM3 fusion protein, which is connected by connecting peptide by steroid 11 beta hydroxylase CYP11B1 and cytochrome P450 BM3 reduction domain.The encoding gene of the fusion protein is transformed into escherichia coli for transformation expression, can efficiently synthesize CYP11B1-BM3 fusion protein, and the prepared fusion protein has higher cytochrome P450 reductase activity and in vitro hydroxylation enzyme activity, can efficiently catalyze hydroxylation reaction, can be applied to the synthesis of cortisol drug.

The invention discloses a method for synthesizing enantio-kaurenoic acid and 16 alpha-hydroxyl enantio-kaurane-19-carboxylic acid based on a recombinant microbiological method. The invention provides genes such as an enantiocopal pyrophosphate synthase gene, a similar enantiokaurene synthase gene, an enantiokaurene oxidase gene, a cytochrome P450 reductase gene, a cytochrome b5 gene, an FPP / GGPP synthase gene and the like which are expressed in a host saccharomyces cerevisiae. The copy number of genes such as an acetyl coenzyme A acyltransferase gene, an HMG coenzyme A synthase gene, a GGPP synthase gene, a glucose-6-phosphate dehydrogenase gene and a 6-phosphogluconate dehydrogenase gene in the saccharomyces cerevisiae is increased to improve the expression or activity; the obtained recombinant saccharomyces cerevisiae is used for producing enantio-kaurenoic acid and 16 alpha-hydroxyl enantio-kaurane-19-carboxylic acid, and a powerful strain and a research basis are provided for biosynthesis of active diterpenoid components.

The application provides a steroid compound 11alpha-hydroxylase and its use. The co-expression vector provided by the application can simultaneously express the steroid compound 11alpha-hydroxylase and cytochrome P450 reductase in Escherichia coli or Mycobacterium, and the genetically engineered bacterial strain containing the co-expression vector can be suitable for 11alpha-hydroxylation of various steroid substrates, has high specificity, and the conversion rate can be up to 90.39%, and the production technology is simple, efficient, controllable, and has important industrial application value.