15 results about "Dammarenediol" patented technology

The application belongs to the technical field of genetic engineering, and discloses a kind of saccharomyces cerevisiae for producing rare ginsenoside by seaweed biomass and a construction method and application thereof.The saccharomyces cerevisiae has the following characteristics: overexpression of agarase, neojuncanohydrolase, hydroxymethylglutaryl coenzyme A reductase, isopentenyl diphosphate delta isomerase, dammaradienol synthase, protopanaxadiol synthase, cytochrome P450 reductase, protopanaxatriol synthase and glycosyltransferase.The application combines enzymolysis of seaweed biomass with fermentation of rare ginsenoside, which not only endows wild-type yeast with the ability to degrade seaweed biomass that it originally does not have, but also effectively improves the yield of squalene and downstream terpenes (rare ginsenoside Rh1) by overexpression of tHMG1 and IDI1.The saccharomyces cerevisiae can effectively utilize seaweed biomass to obtain squalene and rare ginsenoside Rh1, and has the characteristics of simplicity, economy and ecological friendliness, can convert cheap biomass into high-value products, and provides a way for the development of marine bioeconomy.

The invention relates to a dammarendiol synthase mutant and application of the dammarendiol synthase mutant in preparation of beta-amyrin alcohol, and belongs to the technical field of genes. The dammarendiol synthase mutant is subjected to the following amino acid mutation based on a sequence as shown in SEQ ID NO.1: T128L, and other amino acid sequences are kept unchanged; or the dammarendiol synthase mutant is subjected to the following amino acid mutation on the basis of the sequence as shown in SEQ ID NO: 1: T128L, L242M, F253L, F255M, V327L, A377V, V485Q and A493G, and other amino acid sequences are kept unchanged. According to the invention, reverse mutation modification is carried out through the difference sites, so that DDS has a catalytic function of beta-AS, and further deep analysis is carried out on the catalytic difference formed by DDS and beta-AS in the evolution process.

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 disclosure provides compositions and methods related to engineered microbial cells, enzymes, and methods for producing dammarenediol, as well as compounds derived from dammarenediol. Microbial host cells are engineered to express a heterologous biosynthetic pathway that produces dammarenediol, or a derivative thereof. The host cell can optionally express a heterologous uridine diphosphate-dependent glycosyltransferase (UGT) enzyme producing natural or non-natural glycosylated forms of dammarenediol, protopanaxadiol or protopanaxatriol.

The invention discloses recombinant saccharomyces cerevisiae for producing protopanoxadiol by metabolizing glycerol and a construction method thereof, and the construction method comprises the following steps: introducing a dammarendiol synthase DS gene expression cassette, a PPD synthase and reductase fusion protein PPDS-ATR1 gene expression cassette and a squalene oxidase ERG1 gene expression cassette into saccharomyces cerevisiae to obtain a recombinant bacterium 1; introducing a glycerol transport protein CjFPS1 gene expression cassette, a glycerol dehydrogenase OpGDH gene expression cassette, a dihydroxy acetone kinase DAK1 gene expression cassette and a 3-hydroxy-3-methylglutaryl coenzyme A reductase HMGR gene expression cassette into the recombinant bacterium 1 to obtain a recombinant bacterium 2; introducing citric acid lyase AnACLa and AnACLb gene expression cassettes and citric acid transporter protein YHM2 and CTP1 gene expression cassettes into the recombinant bacterium 2 to obtain a recombinant bacterium 3; experiments prove that the yield of the recombinant bacteria 2, 3PPD is 124 mg / L and 220 mg / L.

The invention relates to a beta-amyrin synthase mutant and application of the beta-amyrin synthase mutant in preparation of dammarendiol, and belongs to the technical field of genes. The beta-amyrin alcohol synthase mutant is subjected to the following amino acid mutation on a sequence as shown in SEQ ID NO: 75: M240L, and other amino acid sequences are kept unchanged; or M240L, Q483V, and other amino acid sequences are kept unchanged; the amino acid sequences are M240L, L251F, M253F and Q483V, or M240L, L251F, M253F and Q483V, and other amino acid sequences are kept unchanged; the amino acid sequences are M240L, L251F, M253F, L325V, Q483V and G491A, or M240L, L251F, M253F, L325V, Q483V and G491A, and other amino acid sequences are kept unchanged; the amino acid sequence of the gene can be L128T, M240L, L251F, M253F, L325V, V375S, Q483V or G491A, or other amino acid sequences are kept unchanged. According to the invention, mutation of key amino acids is determined through site-directed mutagenesis and site-directed mutagenesis of beta-AS, so that beta-AS has the function of catalytically generating dammarendiol.

The invention relates to a recombinant saccharomyces cerevisiae capable of producing 23-OH-Dammarenediol II and 25-OH-Dammarenediol II at the same time, and a construction method of the recombinant saccharomyces cerevisiae. The method comprises the following steps: firstly, introducing ERG1, ERG9, ERG20, tHMG1, synPgDDS, synBmCPR1 and a leucine screening tag LEU2 as well as a promoter and a terminator which are derived from yeast into a BY-MVA recombinant bacterium, so as to obtain a recombinant bacterium DM-BmCPR capable of producing dammarendiol; and introducing SynBmCYP068, which is from bacopa monnieri and is subjected to codon optimization, and a Lys screening tag into the DM-BmCPR strain together, so as to obtain the recombinant strain DBmCYP068. The invention also provides a basis for further functional characterization of the downstream modification enzyme gene P450 in the bacopa monnieri biosynthetic pathway.

The invention relates to a gynostemma pentaphylla oxidized squalene cyclase gene GpOSC2 and application thereof, and belongs to the technical field of genes. The amino acid sequence of the gynostemma pentaphylla oxidized squalene cyclase GpOSC2 is as shown in SEQ ID NO. 1. A coding sequence for coding the gynostemma pentaphylla oxidized squalene cyclase GpOSC2 gene is a nucleotide sequence as shown in SEQ ID NO. 2. According to the present invention, the gene is adopted to carry out fermentation so as to obtain Dammarenediol-II, lupeol, lanosterol, cycloartenol and alpha-amyrin alcohol, such that the application prospect is good, and the promotion and the application are easy.

The invention belongs to the technical field of genetic engineering, and discloses saccharomyces cerevisiae for producing rare ginsenoside by utilizing seaweed biomass as well as a construction method and application of the saccharomyces cerevisiae. The saccharomyces cerevisiae has the following characteristics: overexpression agarase, neoagarobiose hydrolase, hydroxymethyl glutaryl coenzyme A reductase, isopentenyl diphosphate delta isomerase, dammarendiol synthase, protopanoxadiol synthase, cytochrome P450 reductase, protopanaxatriol synthase and glycosyl transferase. According to the invention, seaweed biomass enzymolysis and rare ginsenoside fermentation are combined, so that wild yeast is endowed with the capability of degrading seaweed biomass, which does not exist originally; the overexpression of tHMG1 and IDI1 effectively increases the yield of squalene and downstream terpenoids (rare ginsenoside Rh1). The Saccharomyces cerevisiae can effectively utilize seaweed biomass to obtain squalene and rare ginsenoside Rh1, does not have the characteristics of simplicity, economy and eco-friendliness, can convert cheap biomass into products with high added value, and provides a way for developing marine biological economy.

The application discloses a method for producing dammaradien-diol-II glycoside 3beta-O-Glc-DM and a construction method of a recombinant bacterium, the recombinant bacterium obtained by the method, and application of the recombinant bacterium in preparation of 3beta-O-Glc-DM.

The invention discloses a recombinant saccharomyces cerevisiae engineering bacterium which carries a gene expression cassette containing a hydroxymethyl glutaryl-coenzyme A reductase coding gene and an isoprene synthetase coding gene, and discloses the recombinant saccharomyces cerevisiae engineering bacterium which is constructed by adopting the engineering bacterium and is used for producing dammarendiol. The invention discloses a dammarendiol gene carrier which carries a gene expression cassette containing a dammarendiol synthetase gene, and further discloses a method for producing dammarendiol. According to the saccharomyces cerevisiae engineering strain constructed through synthetic biology and genetic engineering technical means, the dammarendiol synthetase can be efficiently recombined and expressed, then a precursor, namely the dammarendiol, of ginsenoside protopanoxadiol is synthesized, and a foundation is laid for finally achieving ginsenoside biosynthesis and large-scale industrial production.

The invention relates to a method for preparing dammarendiol in nicotiana benthamiana and teasel, and belongs to the technical field of biology. The method comprises the following steps: inserting a target gene into a transient expression vector to obtain a recombinant vector; the target gene is as shown in SEQ ID NO. 1; the method comprises the following steps: by taking leaves of seedlings of nicotiana benthamiana and teasel as materials, infecting GV3101 / pEAQ-HT-DDS agrobacterium liquid, culturing, and extracting the cultured plant leaves, thereby obtaining dammarendiol. According to the method, tobacco and teasel leaves are infected by using an agrobacterium tumefaciens vacuum infiltration method, and a safe and efficient transient expression system which is easy for large-scale production of dammarendiol is established in the tobacco and teasel leaves.

The invention relates to a dammarendiol derivative as well as a preparation method and application thereof, and belongs to the technical field of biochemistry. The structure of the dammarendiol derivative is shown as a formula (1). # imgabs0 #, formula (1); in formula (1), R is # imgabs 1 # or # imgabs 2 #. Pharmacological experiments prove that the dammarendiol derivative has an obvious anti-tumor effect, has a better inhibition effect on human breast cancer cells, colon cancer cells, ovarian cancer cells and gastric cancer cell lines than positive control drugs, and can be used as a lead compound or a candidate compound for development of anti-tumor drugs.

The application discloses a dammaradiene synthase and application thereof, and belongs to the technical field of bioengineering.The application aims to provide a protein with dammaradiene synthase and a key amino acid site capable of transforming a triterpene cyclase OSC protein into a catalytic active site for synthesizing dammaradiene.The application provides a dammaradiene synthase, and the amino acid sequence of the dammaradiene synthase is shown as SEQ ID NO.3.It is proved by a site-directed mutation experiment that the mutation of Y260N of the homologous gene of the Panax notoginseng dammaradiene synthase OeOSC14 can make the OSC catalyze the efficient synthesis of dammaradiene from squalene.

The invention belongs to the technical field of medicines, and provides a preparation method of borneol alcohol ketone and dammarendiol II as well as intermediates of the borneol alcohol ketone and the dammarendiol II. The preparation method comprises the following steps: mixing protopanoxadiol with a first additive and an anhydride compound, and reacting to obtain an intermediate 1; mixing the intermediate 1 with anhydrous potassium carbonate and a solvent for reaction to obtain an intermediate 2; mixing the intermediate 2 with a second additive and an oxidizing agent for reaction to obtain an intermediate 3; mixing the intermediate 3 with an alkaline compound and a mixed solvent for reaction to obtain a dammarendiol II intermediate; mixing the dammarendiol II intermediate with ethylene glycol, hydrazine hydrate and KOH for reaction to obtain dammarendiol II; and mixing the dammarendiol II with a third additive and an oxidizing agent, and reacting to obtain the borneol alcohol ketone. The method is simple, efficient, mild in condition and short in time consumption, high-purity borneol alcohol ketone, dammarendiol II and intermediates thereof are prepared, and industrial production of the products is greatly promoted.