49 results about "Cytochrome Reductases" patented technology

The present invention provides several methods for biological production of para-hydroxycinnamic acid (PHCA). The invention is also directed to the discovery of new fungi and bacteria that possess the ability to convert cinnamate to PHCA. The invention relates to developing of a new biocatalyst for conversion of glucose to PHCA by incorporation of the wild type PAL from the yeast Rhodotorula glutinis into E. coli underlining the ability of the wildtype PAL to convert tyrosine to PHCA. The invention is also directed to developing a new biocatalyst for conversion of glucose to PHCA by incorporation of the wildtype PAL from the yeast Rhodotorula glutinis plus the plant cytochrome P-450 and the cytochrome P-450 reductase into E. coli. In yet another embodiment, the present invention provides for the developing of a new biocatalyst through mutagenesis of the wild type yeast PAL which possesses enhanced tyrosine ammonia-lyase (TAL) activity.

The invention provides a method of enhancing the effectiveness of pesticides, as well as pesticidal formulations. Furthermore, it also provides the means for the de novo rational design of pesticides. The present invention also relates to a method of screening agents for potential use in insecticides, particularly against mosquitoes.

Provided is a nicotinamide adenine dinucleotide (NADPH)-cytochrome P450 reductase and a use thereof. Also provided is a polynucleotide encoding the NADPH-cytochrome P450 reductase, an expression vector and a host cell which express the NADPH-cytochrome P450 reductase, and a method for producing the protopanoxadiol.

A bacterial cell containing a functional cytochrome P450 monooxygenase system, said cell comprising a genetic construct capable of expressing a cytochrome P450 and genetic construct capable of expressing, separately from said cytochrome P450, a cytochrome P450 reductase, wherein each of the cytochrome P450 and the cytochrome P450 reductase have at their N-terminus a bacterial signal peptide. The cytochrome P450 and the cytochrome P450 reductase may be encoded by different constructs or the same construct. A bacterial cell containing a cytochrome P450 comprising a genetic construct encoding, and capable of expressing, said cytochrome P450 wherein the cytochrome P450 has, at is N-terminal, a bacterial signal peptide. The bacterial cells are useful as, for example, bioreactors, in drug testing and mutagenicity testing and as a source of cytochrome P450.

The invention provides a method for synthesizing 3-O-glucose-based oleanolic acid and cellobiose oleanolic acid by using saccharomyces cerevisiae engineering bacteria, and belongs to the field of bioengineering. The method comprises the following steps: synthesizing a codon optimized P450 cytochrome monooxygenase gene, a cytochrome reductase gene and a UDP-glucosyltransferase gene by a chemical method; constructing corresponding gene expression boxes by combining a saccharomyces cerevisiae promoter with a terminator; constructing gene expression vectors by a DNA (Deoxyribonucleic Acid) klenow fragment assembling method, and importing the gene expression vectors into the saccharomyces cerevisiae capable of producing beta-amyrin. Direct synthesis of the 3-O-glucose-based oleanolic acid and the cellobiose oleanolic acid serving as plant secondary metabolites in the saccharomyces cerevisiae is realized for the first time; in addition, two synthesized compounds can span cytomembrane of the saccharomyces cerevisiae engineering bacteria, and a downstream separation and extraction process is simplified, so that a new idea is provided for producing pentacyclic triterpene compounds with low water solubility and difficulty in spanning membranes by using the saccharomyces cerevisiae. The method is simple in process and can be used for producing the 3-O-glucose-based oleanolic acid and the cellobiose oleanolic acid by fermenting.

The invention discloses a recombinant Yarrowia lipolytica for producing dammarendiol and protopanoxadiol and application. The recombinant Yarrowia lipolytica is constructed by the following steps: introducing an optimized dammarendiol synthase encoding gene DS expression cassette, an optimized protopanoxadiol synthase encoding gene PPDS expression cassette and an optimized encoding gene expressioncassette of cytochrome-NADPH-reductase 1 into Yarrowia lipolytica so as to obtain a recombinant yeast 2, and cutting 1500 nucleotides from the 5'end of a gene HMG1 for encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase to obtain a truncated gene tHMG1 for encoding the 3-hydroxy-3-methylglutaryl coenzyme A reductase; and introducing the truncated gene tHMG1 into recombinant yeast 2 to obtainrecombinant yeast 3. Experiments prove that the yield of dammarendiol and protopanoxadiol is increased by acquiring the recombinant Yarrowia lipolytica through a homologous recombination method. Themethod disclosed by the invention provides a basis for artificially synthesizing the dammarendiol and the protopanoxadiol.

The invention discloses a microbial fermentation method for synthesizing triterpene compounds, namely a distillers yeast recombination strain SY2 containing cucurbitane dienol synthetase SgCbQ, P450 CYP87D18 and cytochrome reductase CPR, in the metabolism route of mogrol, which takes dextrose as a substrate to synthesize three new products including 11-oxygen-cucurbitane dienol, 11-hydroxy-cucurbitane dienol and 11-oxygen-24, 25-epoxy-cucurbitane dienol. The invention further discloses a metabolism route for synthesizing mogrol through cucurbitane dienol, namely the cucurbitane dienol is catalyzed through P450 CYP87D18 and synthesized into 11-oxygen-cucurbitane dienol, 11-oxygen-cucurbitane dienol is catalyzed through enzyme and synthesized into 11-hydroxy-cucurbitane dienol and 11-oxygen-24, 25-epoxy-cucurbitane dienol, and 11-hydroxy-cucurbitane dienol and 11-oxygen-24, 25-epoxy-cucurbitane dienol can be synthesized into the mogrol through other P450 or epoxide hydrolase. The invention further discloses a novel thought for obtaining more glycosylation products by catalyzing new products through glycosyl transferase. The method has significance for determining the synthesis route of mogrol and synthesizing relevant intermediate metabolites and glycosylation products through the microbial fermentation method.

The invention relates to cytochrome P450, a nicotinamide adenine dinucleotide-cytochrome P450 reductase and application thereof. The invention discloses the set of new cytochrome P450 (P450 GP01 and P450 GP02) which performs the effect in the terpenoid hydroxylation catalysis and synthesis of Ganoderma triterpenes, and the nicotinamide adenine dinucleotide-cytochrome P450 reductase (GLCPR) which completes hydroxylation catalysis by cooperating with the cytochrome P450. In specific, the cytochrome P450 GP01 can specifically and efficiently catalyze the hydroxylation of C-23 site of a tetracyclic triterpene compound substrate. The cytochrome P450 GP02 performs hydroxylation om ganoderic acid DM and ganoderic acid TR to generate various kinds of products. The GLCPR can be paired with the cytochrome P450, and assists the cytochrome P450 to perform the hydroxylation activity.

A highly efficient method for catalyzing the synthesis of sanguinarine and chelerythrine, comprising: from the known protopine-6-hydroxylase gene, dihydrobenzophenanthridine oxidase gene and cytochrome P450 reductase gene, by means of heterologous expression and result comparison and analysis, screening out an optimal gene having high expression efficiency, then performing codon optimization on the selected optimal gene; constructing the optimal gene sequence on an expression vector, then transferring same into yeast engineering bacteria for transformation so as to obtain recombinant yeast engineering strains; finally, feeding the recombinant yeast engineering bacteria by using a leaf raw material liquid precursor of Macleaya cordata for fermentation, so as to obtain a product. The invention improves the enzyme catalytic efficiency of sanguinarine and chelerythrine in various aspects such as gene level and fermentation process; the leaf raw material liquid, which is not a traditional medicinal part of Macleaya cordata, is directly used for fermentation with the engineering bacteria, and protopine and allocryptine having high alkaloid content in leaves are converted into high-value sanguinarine and chelerythrine to achieve the comprehensive utilization of Macleaya cordata resources.