24 results about "Cytochrome Reductases" patented technology

The invention discloses a mesophilic carotenoid-producing recombinant yarrowia lipolytica strain as well as a construction method and application thereof, the recombinant strain exogenously expresses mesophilic original fatty acid desaturase (FAD3) and isocitrate dehydrogenase (IDH2) mutants from phaffia rhodozyma, and efficient synthesis under the condition of 30 DEG C is realized; secondly, a metabolic pathway of the recombinant strain is optimized based on a metabolic flow push-pull strategy, and isopentenyl pyrophosphate isomerase (IDI), farnesyl pyrophosphate synthase (ERG20), plant alkene dehydrogenase (CarB) and bifunctional plant alkene synthase (CarRP) are overexpressed, so that the flux of a precursor is increased; finally, gene multi-copy of astaxanthin synthase (crtS) and cytochrome P450 reductase (crtR) from phaffia rhodozyma is carried out, and the carotenoid production capacity of yarrowia lipolytica is further improved. The construction method of the mesophilic yarrowia lipolytica is simple, efficient synthesis of carotenoid can be better achieved at the higher temperature, the fermentation cost is greatly reduced, and the industrial production potential is achieved.

The invention relates to an application of a cytochrome P450 reductase 10273 inactivating agent in construction of an aspergillus terreus engineering strain for high yield of emodin or a derivative thereof. The invention further relates to aspergillus terreus engineering bacteria for high yield of emodin or the emodin derivatives and a construction method of the aspergillus terreus engineering bacteria. By knocking out the cytochrome P450 reductase gene 10273, the activation of anthraquinone methyl hydroxylase 4804 is blocked in aspergillus terreus, so that the hydroxylation pathway of the emodin is blocked, the purity of the emodin and the emodin-3-methyl ether in aspergillus terreus cells is improved, the production cost of the emodin and the emodin-3-methyl ether is reduced, and the method is suitable for industrial production. And a foundation is provided for the microbial industrial synthesis of the emodin and the emodin-3-methyl ether.

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 present invention belongs to the field of biotechnology, and particularly relates to a bionic synthesis engineering bacterium of compound Danshen and its application. The engineering bacterium is obtained by introducing cytochrome P450 reductase, tyrosine hydroxylase and D-lactate dehydrogenase of Lactobacillus pentosus on the basis of a Saccharomyces cerevisiae starting bacterium that produces protopanaxadiol, cryptotanshinone diene and borneol. The obtained Saccharomyces cerevisiae genetic engineering bacterium can simultaneously synthesize the active ingredients of compound Danshen preparations, namely protopanaxadiol, cryptotanshinone diene, danshensu and borneol, and has application value.

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 biology, and particularly relates to compound salvia miltiorrhiza biomimetic synthesis engineering bacteria and application thereof. The engineering bacterium is obtained by introducing cytochrome P450 reductase, tyrosine hydroxylase and D-lactic dehydrogenase of lactobacillus pentosus on the basis of starting bacteria of saccharomyces cerevisiae for producing protopanoxadiol, hypotanshinone diene and borneol. The obtained saccharomyces cerevisiae genetically engineered bacterium can be used for simultaneously synthesizing active ingredients of protopanoxadiol, hypotanshinone diene, tanshinol and borneol of a compound salvia miltiorrhiza preparation, and has application value.

The invention discloses a method for improving the production of ursolic acid and oleanolic acid by yeast cerevisiae, and particularly relates to fermentation production using recombinant yeast cerevisiae. The recombinant yeast at least expresses amyrin synthase CrMAS, amyrin C-28 oxidase CrAO, and a cytochrome NADPH reductase AtCPR1, knocks out malate synthase MLS1, and enhances the expression of alcohol dehydrogenase ADH2, acetate-CoA ligase 1ACS1, aldehyde dehydrogenase ALD6, squalene epoxidase ERG1, and amyrin synthase CrMAS.

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.

The present application describes enzymes (e.g., cucurbitacinadienol synthase (CDS), UDP-glycosyltransferase (UGT), C11 hydroxylase, epoxidase (EPH), squalene epoxidase, and / or cytochrome P450 reductase), host cells expressing the enzymes, and methods of producing mogroside precursors, mogroside, and / or mogroside using such host cells.

The invention provides a catalyst for catalyzing bioconversion of steroid drugs from steroids and a preparation method of the catalyst. According to the present invention, the C21 hydroxylase CsCYP21A or the combination of the C21 hydroxylase CsCYP21A and the cytochrome P450 reductase CPRCS is adopted as the catalyst, the recombinant expression vector is constructed, the pichia pastoris KM71H is adopted as the expression host, the heterologous expression of the C21 hydroxylase CsCYP21A is achieved, and the efficient hydroxylation conversion of a variety of different steroid compounds C21 is successfully achieved.

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 present invention discloses a heme oxygenase mutant and its multi-enzyme coupling system for the efficient synthesis of bilirubin, belonging to the field of biocatalytic engineering. The present invention provides a heme oxygenase mutant that catalyzes the synthesis of biliverdin from heme; couples biliverdin reductase to catalyze the synthesis of bilirubin from biliverdin; introduces cytochrome P450 reductase to provide electrons; carbon monoxide dehydrogenase removes the competitive binding of by-products to the substrate heme; formate dehydrogenase is used for in-situ regeneration of coenzymes, and bilirubin is efficiently synthesized by a one-pot method. The heme oxygenase mutant of the present invention couples the above two core enzymes with three foreign auxiliary enzymes to construct recombinant Escherichia coli, which catalyzes heme without adding exogenous NADPH, and the bilirubin yield is as high as 1611 mg / L within 4 h, which is the highest reported bilirubin synthesis level with the shortest time. This technology not only removes resource limitations, but also has a simple operation method and a high yield, realizing the efficient and green synthesis of bilirubin.

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 provides an Escherichia coli engineering bacterium for improving the yield of astaxanthin by using yhaK as well as a construction method and application of the Escherichia coli engineering bacterium. The engineering bacterium comprises a recombinant plasmid containing a yhaK gene, a crtS gene for coding astaxanthin synthetase, a crtR gene for coding cytochrome reductase, a crtYB gene for coding lycopene cyclase and a gene segment for coding farnesyl / geranyl pyrophosphate synthase IspA, the escherichia coli engineering strain ELZSS02 E.coli Bl21 (DE3): pTrc low-MvaE-MvaS / pET-RPlyc-crtS / pACYC-crtR-crtYB-IspA-yhaK is finally obtained, compared with wild escherichia coli, the expression quantity of MEP pathway genes in the engineering strain is remarkably increased, the yield of astaxanthin is also remarkably increased, and the escherichia coli engineering strain has good application value.

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 present invention relates to a genetically engineered bacterium with high gibberellin GA3 production, a construction method and an application. The present invention enhances the dissolved oxygen level in the gibberellin synthesis process and enhances the supply of the electron transport chain by enhancing the expression of a gene encoding cytochrome P450 reductase CPR and a gene encoding Vitreoscilla hemoglobin VHB in Fujikura Gibberella. The recombinant Fujikura Gibberella with high gibberellin GA3 production provided by the present invention has enhanced oxidation capacity in the late stage of metabolism compared with the wild type, improved synthesis capacity in the metabolic process, and increased accumulation of hyphae, and can better utilize nutrients for the production of gibberellin GA3. The strain with the best performance after the transformation has a level of gibberellin GA3 production in fermentation increased from 2g / L to 3.45g / L compared with the starting strain, significantly improving the gibberellin GA3 production capacity, and the increase in hyphae can increase the time-space yield of gibberellin GA3 synthesis, and can be applied to large-scale gibberellin GA3 production.

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.