14 results about "Protopanaxadiol" 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 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 provides a conversion method of rare ginsenoside Rg5. According to the method, through a vacuum permeation technology, gas in cell gaps of ginseng roots is removed in a vacuum environment, a permeation barrier of cell walls is weakened, a micro-channel is formed, and then a mixed solution of methanol and n-butyl alcohol is injected for primary extraction; after the normal pressure is recovered, the mixed solution can be driven by differential pressure to quickly fill a micro-channel and be better dissolved with the ginseng roots, a more efficient extraction environment is created, extraction is carried out under the low-temperature condition of 30-50 DEG C, thermal degradation of PPD can be effectively avoided, meanwhile, dissolution of impurities such as polysaccharide and protein is reduced, and the adverse effect of the impurities on subsequent conversion is reduced; finally, cellulose nanocrystals and a hydrochloric acid solution are added, on the basis that extraction is further promoted, protopanoxadiol saponin can be efficiently and directionally converted into rare ginsenoside Rg5, meanwhile, generation of an isomeride Rk1 of the rare ginsenoside Rg5 is reduced, and then the yield and purity of the Rg5 are remarkably improved.

This invention discloses a compound preparation for treating vascular dementia and its preparation method. The active pharmaceutical ingredient consists of protopanaxadiol and gallic acid in a 1:1 mass ratio, and the dosage form is liposomes. The preparation method includes extraction, mixing, liposome preparation, and optional freeze-drying steps: protopanaxadiol and gallic acid are extracted and purified from ginseng rhizome and Terminalia chebula fruit, respectively. After mixing, they are combined with phospholipids and cholesterol to prepare a lipid membrane. The membrane is then homogenized by hydration, low-temperature ultrasound, and high-pressure microfluidic jet to obtain a homogeneous liposome suspension, which can be further freeze-dried into a freeze-dried powder. The 1:1 composition of protopanaxadiol and gallic acid targets multiple points on the pathological mechanism of vascular dementia, showing significant therapeutic effects and broad application prospects. This preparation uses a liposome dosage form to improve the bioavailability of the active ingredient, the preparation process parameters are controllable, the active ingredient has high purity, the liposomes have uniform particle size, high encapsulation efficiency, and good stability.

The invention discloses application of a ginsenoside and panoxadiol composition in preparation of a preparation for improving intestinal microbiota, and relates to the technical field of plant extract application, the composition at least comprises ginsenoside Rb3 and 20 (S)-protopanoxadiol which are used after being mixed according to the mass ratio of 1: (1-2); the composition can reverse damaged intestinal flora of high-fat diet mice, ginsenoside Rb3 and 20 (S)-protopanoxadiol are combined for application, the lipid metabolism related gene expression level is adjusted through the synergistic effect, the intestinal microbial flora composition of the high-fat diet mice can be effectively recovered, and an experimental basis is provided for research of an intestinal liver axis mechanism.

The application discloses a ginsenoside-protopanaxadiol hydrogen sulfide donor derivative, a preparation method and application thereof, and belongs to the technical field of natural medicines and medicinal chemistry. Ginsenoside-protopanaxadiol is used as a lead compound, a series of ginsenoside-protopanaxadiol hydrogen sulfide donor derivatives are designed and synthesized, and myocardial protection activities of the synthesized derivatives are tested. The results show that the ginsenoside-protopanaxadiol hydrogen sulfide donor derivative prepared by the application has better cardiac protection activities for treating myocardial ischemia, arrhythmia and myocardial infarction, and has the potential as a cardiac protection drug.

The invention relates to a novel protopanoxadiol derivative as well as a preparation method and application thereof. The novel protopanoxadiol derivative is obtained by carrying out methoxy acetylation reaction on protopanoxadiol. Activity studies show that the compound has a certain inhibition effect on lung cancer, endometrial cancer and gastric cancer cells. Compared with a parent compound protopanoxadiol, the protopanoxadiol has the advantage that the inhibition rate on human lung cancer cells and human gastric cancer cells is obviously improved.

The invention discloses a glycosyl transferase mutant, and belongs to the technical field of gene engineering and enzyme engineering. According to the invention, O-glycosyl transferase UGT73F13 in a wild type UGT73 family derived from Glycyrrhiza uralensis (Glycyrrhiza uralensis) is subjected to semi-rational design mutation from outside to inside, and an excellent amino acid mutant is screened out. The multi-site mutant of the novel O-glycosyl transferase contains five mutants, wherein V403 is mutated into S, C274 is mutated into T, C269 is mutated into N, S87 is mutated into A, and L84 is mutated into T. The catalytic efficiency of the obtained novel mutant is improved by 9.29 times compared with that of a wild type, the thermal stability (T1 / 2 value) is improved by 8.4 times, the substrate conversion rate of protopanoxadiol (PPD) is greater than or equal to 95%, and the C-3 hydroxyl glycosylation specificity reaches 100%. The invention provides a unique biocatalyst and a conversion process for the preparation of 20 (S)-ginsenoside Rh2 with high added value.

The invention discloses a method for preparing rare ginsenoside 20 (S)-Rg3 by directionally converting panax notoginseng stem and leaf saponin based on multienzyme synergistic catalysis, and belongs to the technical field of enzyme engineering. The method comprises the following steps: firstly, generating 20 (S)-Rg3 by using multifunctional xylosidase Tpexy3 and glucosidase Tpebgl3; further, glucosidase Dth3 and glucosidase Tpebgl1 are used for efficiently converting the saponin of the non-Rg3 into the protopanoxadiol; finally, PPD is used as a substrate, and directional synthesis of 20 (S)-Rg3 is achieved through a three-enzyme cascade reaction system composed of UGT mutants MuUGT73F13, UGT94Q2 and sucrose synthase GmSUS. According to the method, the resource advantages of the PPD type saponin in the pseudo-ginseng stems and leaves are fully utilized, the raw material utilization rate and the product added value are remarkably improved, and a feasible path is provided for green and efficient production of the rare ginsenoside.

The application discloses a glycosyltransferase, which is a parent of wild-type glycosyltransferase described in SEQ ID NO. 1, and comprises the following mutation points: the 125th lysine is mutated into arginine (K125R), the 172nd alanine is mutated into phenylalanine (A172F), the 294th glycine is mutated into cysteine (G294C), and the 311th valine is mutated into cysteine (V311C), and application of the glycosyltransferase in preparation of non-natural proto-ginsenoside. The application rationally designs and constructs multiple groups of mutant UGT109A1 of UGT109A1 by analyzing the structure and evolutionary conservation of the enzyme molecule UGT109A1, and obtains the mutant with improved catalytic performance under the concentration of 30% DMSO through screening, and the glycosylation efficiency of the mutant is relatively improved by 72% compared with the wild-type enzyme. Based on the wide substrate spectrum of UGT109A1, the obtained mutant enzyme with improved glycosylation efficiency under high DMSO concentration provides a new enzyme for green and efficient preparation of functional glycosides.

The application belongs to the technical field of biological medicine, and particularly relates to an application of a proto-prosolan in preparation of a medicine for treating acute kidney injury, especially in preparation of a medicine for preventing and treating sepsis-induced acute kidney injury. Animal experiments show that the proto-prosolan can significantly reduce necrosis, vacuolar degeneration and interstitial edema of renal tubular epithelial cells, and obviously improve structural damage of the kidney, thereby providing an effective approach for preventing and treating acute kidney injury.

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.

The application discloses application of a glycosyltransferase mutant in preparation of active ginsenoside, and the mutant is obtained by mutating the 12th residue in the amino acid sequence of glycosyltransferase Yjic into another amino acid residue; the amino acid sequence of the glycosyltransferase Yjic is shown in SEQ ID NO: 2. With the glycosyltransferase mutant constructed in the application and a sucrose synthase AtSuSy double-enzyme coupling catalytic system, active ginsenoside F12 can be directly prepared in one step with protopanaxadiol as a substrate, the substrate conversion rate is high, the product is relatively single, the subsequent separation step is reduced, and the product purification process is simplified. Ginsenoside F12 has multiple medicinal activities such as inhibiting tumor cells and improving immunity, and therefore has great prospects in development of new drugs.

This invention relates to glycosyltransferase mutants and their application in the enzymatic preparation of rare ginsenosides. (The text then abruptly shifts to a discussion of glycosyltransferase UGT.) BL 1. Site-directed mutagenesis was performed, and the resulting mutants can be used to efficiently and directionally prepare rare ginsenoside Rh1. The regioselectivity of the enzymatic preparation of rare ginsenoside Rh1 reached 96.26% (conversion rate 95.56%). Glycosyltransferase UGT BL 1. Some other mutants can also be used to directionally synthesize non-natural ginsenoside protopanaxadiol-3- O -β-glucan (3- O -β-Glc-PPT), enzymatic preparation of non-natural ginsenosides 3- O The regioselectivity of -β-Glc-PPT reached 84.83% (conversion rate 98.13%). This invention utilizes the glycosyltransferase UGT. BL The mutant of 1 and sucrose synthase At The SuSy dual-enzyme coupled catalytic system was used to prepare Rh1 or 3- O -β-Glc-PPT, and significantly increased the levels of rare ginsenosides Rh1 and 3- by using a fed-batch method. O The product concentration level of -β-Glc-PPT simplifies the preparation process; the product is relatively homogeneous, which helps reduce subsequent separation steps and streamlines the product purification process. 3- O -β-Glc-PPT can be used to prepare drugs for the treatment of diseases such as tumors, and has broad prospects for new drug development.