30 results about "Dammarenediol" patented technology

The invention discloses recombinant saccharomyces cerevisiae for producing dammarenediol and protopanoxadiol using xylose and a construction method. The construction method comprises the steps of replacing a promoter of a saccharomyces cerevisiae xylulokinase gene XKS1 with a promoter PFBA1 by virtue of a homologous recombination method, introducing xylose reductase XYL1 and a xylitol dehydrogenase XYL2 expression cassette, increasing the activities of transketolase TKL1 and transaldolase TAL1 so as to obtain recombinant bacteria 1, introducing a farnesyl-diphosphate farnesyltransferase gene ERG9, a squalene monooxygenase gene ERG1 and a dammarenediol synthase gene DS into the recombinant bacteria 1 so as to obtain recombinant bacteria 2, and introducing a nicotinamide adenine dinucleotide-hydroxymethylglutaryl coenzyme A reductase gene NADH-HMGr, farnesyl diphosphatesynthase ERG20 and a protopanoxadiol synthase-cytochrome P450 reductase fusion protein gene PPDS-ATR1 into the recombinant bacteria 2, so as to obtain recombinant bacteria 3. According to the recombinant saccharomyces cerevisiae, dammarenediol and protopanoxadiol can be artificially synthesized by virtue of xylose.

The invention relates to a biosynthesis method of dammarenediol and provides a producing strain of the dammarenediol. A pichia pastoris engineering strain, which can recombine and express a dammarenediol synthetic enzyme with high efficiency, is constructed through a genetic engineering technical means, a precursor for synthesizing protopanaxadiol, namely the dammarenediol is further synthesized, and a foundation is finally laid for biosynthesis and large-scale industrial production of the protopanaxadiol.

The invention discloses an escherichia coli gene engineering strain for synthesizing Dammar enediol and a construction method. The escherichia coli gene engineering strain for synthesizing Dammar enediol contains a squalene synthase gene with 26 amino acid residues at the truncated C end, a 2,3-oxidized squalene synthase gene, a cytochrome-NADPH-reductase 1 gene in arabidopsis with 45 amino acid residues at the truncated C end, and a Dammar enediol synthase gene. The construction method disclosed by the invention is used for successfully constructing the escherichia coli gene engineering strain for synthesizing the Dammar enediol. Compared with other host bacteria, the escherichia coli gene engineering strain disclosed by the invention has the advantages that the way of synthesizing the Dammar enediol by escherichia coli is easy to regulate and control, and metabolic engineering transformation is facilitated, so that a foundation is laid for fermentation and production of the escherichia coli of Dammar cucuribitane saponin.

The invention discloses fusion protein capable of improving dammarenediol conversion efficiency, a construction method and application. The construction steps comprise: excising first 138 bases at 5' end of arabidopsis thaliana cytochrome-NADPH-reductase 1 gene AtCPR1, so as to obtain a sequence shown as SEQ ID NO. 2; (2) removing a termination codon TAA at 3' end of ginseng protopanaxadiol synthase gene PPDS with a sequence shown as SEQ ID NO. 3, and connecting with the base sequence which at 5' end of the sequence shown as SEQ ID NO. 2 and used for coding polypeptide GSTSSGSG, so as to construct a gene member of fusion protein; and (3) connecting the gene member of the fusion protein with saccharomyces cerevisiae cell endogenous promoter and terminator, so as to construct a fusion protein gene expression cassette, and transforming into saccharomyces cerevisiae cell for expression. The fusion protein constructed by the method is capable of improving the conversion rate of dammarenediol to protopanaxadiol.

The invention relates to a protopanoxadiol biosynthesizing method and provides a production bacterial strain of the protopanoxadiol. According to the protopanoxadiol biosynthesizing method, a pichia pastoris engineering strain constructed through a genetic engineering technological means can be used for efficiently co-expressing dammarendiol synthetase and P450 cytochrome synthetase, and further the protopanoxadiol is catalyzed and synthesized. The protopanoxadiol biosynthesizing method lays a foundation for large-scale industrial production of the protopanoxadiol.

The invention discloses a fusion protein capable of increasing the conversion efficiency of dammarendiol, a construction method and an application. The construction method comprises the following steps: (1) excising the front 138 basic groups of the end 5' of the cytochrome-NADPH-reductase 1 gene AtCPR1 in arabidopsis to obtain a sequence shown in SEQ ID NO. 2; (2) removing the termination codon TAA of the end 3' of the synthase gene PPDS of protopanoxadiol in ginseng shown in SEQ ID NO. 3, and connecting with a base sequence of an encoding polypeptide GSTSSG for the end 5' of a sequence shown in SEQ ID NO. 2 to construct a genetic element of the fusion protein; (3) connecting the genetic element of the fusion protein with the internal promoter and the terminator of a saccharomyces cerevisiae cell to construct a genetic expression kit of the fusion protein, and transforming to enter the saccharomyces cerevisiae cell for expression. The fusion protein constructed by the method disclosed by the invention is capable of increasing the conversion efficiency of converting from dammarendiol to protopanoxadiol.

The invention discloses a cultivation method of a novel dammarane-type ginsenoside-containing paddy rice variety and belongs to the field of genetic engineering. The breeding method is characterized in that a full-length cDNA of a gene of a dammarenediol synthetase as a key enzyme of dammarane-type ginsenoside biosynthesis is constructed to double-T-DNA plant expression vectors; the constructed product is subjected to genetic transformation in paddy rice; and through screening cultivation, the novel dammarane-type ginsenoside-containing paddy rice variety is obtained. A test proves that the novel dammarane-type ginsenoside-containing paddy rice variety comprises 0.020% of 20(S)-protopanoxadiol (PPD) and 0.173% of 20(S)-protopanaxatriol (PPT). The cultivation method can be used for cultivation of high-quality ginseng rice varieties containing dammarane-type ginsenoside, can improve a health care value of rice as a main food of a human body, can widen a ginsenoside source, can alleviate the scarcity of a ginsenoside drug source, and has a good application prospect.

The invention provides an American ginseng identification DNA hybridization method. An applied probe applied in the American ginseng identification DNA hybridization method is designed on the basis ofthe locus of Single Nucleotide Polymorphism (SNP) of dammarenediol synthase genes of panax genomes. Experiments of the American ginseng identification DNA hybridization method detects that Chinese ginseng, American ginseng and pseudo-ginseng are high in similarity of dammarenediol synthase genes. For identifying American ginseng more accurately, the American ginseng identification DNA hybridization method designs a number of probe sequences and screens out probe sequences capable of accurately identifying American ginseng.

The invention discloses a genetic engineering bacterium for producing protopanaxadiol. The genetic engineering bacterium for producing protopanaxadiol is an engineering bacterium for expressing a squalene synthase gene, a 2,3-oxidosqualene synthase gene, a dammarenediol synthase gene, a protopanaxadiol synthase gene and a nicotinamide adenine dinucleoside phosphate-cytochrome P450 reductase gene in Escherichia coli. The protopanaxadiol is biologically synthesized by using the genetic engineering bacterium, so the genetic engineering bacterium and the method have the advantages of short fermentation time, simple medium, meeting the requirements of modern industry, and facilitation of promotion and application.

The invention discloses a recombinant yarrowia lipolytica strain for producing protopanoxadiol by utilizing xylose and a construction method and application of the recombinant yarrowia lipolytica strain. Yarrowia lipolytica ATCC 201249 is used as an original strain, optimized xylose reductase XYL1, xylitol dehydrogenase XYL2 and an overexpressed endogenous xylulokinase XKS expression cassette areintroduced, and a recombinant strain 1 is obtained; a dammarendiol synthase gene DS, a protopanoxadiol synthase gene PPDS and a cytochrome P450 reductase gene ATR1 are introduced into the recombinantstrain 1 to obtain a recombinant strain 2; a gene tHMG1 for encoding nicotinamide adenine dinucleotide-hydroxymethyl glutaryl coenzyme A reductase, a gene erg9 for squalene synthase and an erg20 for encoding farnesene pyrophosphate synthase in a truncated mevalonic acid pathway are introduced into the recombinant strain 2 to obtain a recombinant strain 3; and the recombinant yarrowia lipolytica isused for producing the protopanoxadiol by using the xylose, and the shake flask yield reaches 80.88 mg/L.

The invention belongs to the field of traditional Chinese medicines, and relates to an application of an Erzhi formula compound in preparation of a drug for preventing and treating benign prostatic hyperplasia (BPH). Specifically, the invention relates to an application of a compound in preparation of a medicine selected from the following medicines: (1) a medicine for treating and/or preventing benign prostatic hyperplasia; (2) a medicine for inhibiting proliferation of human prostatic hyperplasia epithelial cells; or (3) a drug for inhibiting PCNA protein expression and/or inhibiting mRNA expression; wherein the compound is selected from dammarendiol, p-hydroxyphenylethanol, ligustrosidic acid, verbascoside, oleoside methyl ester, luteolin, ABiochanin A, echinocystic acid and echinocystic acid-28-O-beta-D-glucose. The Erzhi formula compound or a combination of the Erzhi formula compound is effective in the treatment and/or prevention of benign prostatic hyperplasia.

The invention discloses a fusion protein capable of improving the dammarendiol conversion efficiency and a construction method thereof. The construction method comprises the following steps: (1) cutting off the front 138 bases at the 5' end of the cytochrome-NADPH-reductase 1 gene AtCPR1 in arabidopsis to obtain the sequence shown by SEQ ID No.2; (2) removing the termination codon TAA at the 3' end of the protopanoxadiol synthase gene PPDS in ginseng shown by SEQ ID No.3, and connecting the termination codon TAA with the base sequence of the coding polypeptide GGG at the 5' end of the sequence shown by SEQ ID No.2 to construct a gene element of the fusion protein; and (3) connecting the gene element of the fusion protein with the endogenous promoter and terminator of saccharomyces cerevisiae cells to construct a gene expression box of the fusion protein, and converting into the saccharomyces cerevisiae cells for expression. The fusion protein disclosed by the invention can improve the conversion rate of dammarendiol to protopanoxadiol.