505 results about "Synthetic enzyme" patented technology

Provided are S-adenosylmethionine synthetase mutants and the use thereof, belonging to the technical field of biological enzyme engineering. Site-directed combinatorial mutations are performed on positions 189, 200, 234, 266 and 371 of S-adenosylmethionine synthetase MAT to obtain a series of mutants with improved enzyme activity, wherein the specific enzyme activity of I189V / V266H and V266H / N371R is increased by more than 10 times relative to the parent. The yields of S-adenosylmethionine synthesized by whole cells of recombinant bacteria expressing mutants I189V / V266H / N371R, V266H / Q234N / N371R and I189V / V266H / Q234N / N371R all reach 1550 mg / L or more. The provided enzyme mutants have broad application prospects in the production of S-adenosylmethionine and the construction of genetically engineered microorganisms for S-adenosylmethionine.

The invention provides a 9-hydroxycamptothecin synthetase and a method for synthesizing the 9-hydroxycamptothecin. The invention discloses the 9-hydroxycamptothecin synthetase for the first time, and establishes a method for producing the 9-hydroxycamptothecin. According to the technical scheme, biological synthesis of the 9-hydroxycamptothecin is achieved, and a new way is provided for producing camptothecin derivatives based on strategies such as synthetic biology.

The invention discloses a double-function flavin adenine dinucleotide synthetase mutant and application, the double-function flavin adenine dinucleotide synthetase is called RFK / FADS for short, and the amino acid sequence of the double-function flavin adenine dinucleotide synthetase is shown as SEQ ID NO.1; and the RFK / FADS mutant is RFK / FADSF55A, RFK / FADSG43Q, RFK / FADSG43Q-F55H, RFK / FADSG43Q-F55N or RFK / FADSG43Q-F55N-F129Y, and the RFK / FADSG43Q-F55N and the FADSG43Q-F55N-F129Y can be used for preparing the According to the genetically engineered bacterium disclosed by the invention, flavine adenine dinucleotide is produced through a simple fermentation process by taking glucose as a substrate under a shake flask aerobic condition, and the yields of the strains ZF02, ZF03, ZF04, ZF05 and ZF06 are respectively increased by 120%, 130%, 166%, 246% and 286% in sequence compared with those of ZF01.

The invention belongs to the technical field of immunotherapy, and particularly relates to a method for improving NK cell functions by targeted inhibition of lactylation. The invention provides a method for improving functions of NK cells by targeted inhibition of lactylation, NK cell lactic acid transporters or modification enzymes in inhibition of the lactylation process are targeted, the modification enzymes are NK cell lactylation modification enzymes Writer or lactic acid coenzyme A synthetase, NK92MI lactylation modification is blocked, so that function depletion of the NK cells is reversed, and the NK cell function is improved. The function of the NK cell can be partially reversed by inhibiting the lactic acid modification, and the expression level of effector molecules of the NK92MI cell can be recovered.

The invention discloses a pyranopyridine compound as well as a preparation method, a pharmaceutical composition and application thereof. The invention provides a compound as shown in a formula (I), a pharmaceutically acceptable salt or a stereoisomer thereof, and the compound has a relatively strong inhibition effect on aldosterone synthetase, almost has no influence on cortisol synthetase, has high selectivity and relatively high safety, and can be used for preventing and / or treating various diseases related to aldosterone. # imgabs0 #

The invention relates to a method for increasing the yield of L-malic acid produced by fermentation by adding ethylene glycol, which comprises the following steps: constructing genetic engineering escherichia coli, and introducing ethylene glycol metabolism genes (overexpression) including glycerol dehydrogenase, lactaldehyde reductase, lactaldehyde dehydrogenase and the like; and overexpression of other genes involving malic acid production, such as malic acid synthase, phosphoenolpyruvate carboxylase and malic acid export protein. The gene engineering strain is an E.coli BW25113 strain of which ldhA, pta, poxB, adhE, pflB, mgsA, frdA, maeA, maeB, yqhD, ycdW and iclR genes are substantially inactivated, and the deleted genes are beneficial to reducing the generation of metabolic byproducts and increasing the accumulation of malic acid. The genetic engineering strain is subjected to fermentation culture by taking glucose and ethylene glycol as carbon sources, and L-malic acid is separated from a culture after culture is finished.

The invention discloses a mutant with improved catalytic activity of S-adenosylmethionine synthetase and application of the mutant, and belongs to the technical field of biological enzyme engineering. According to the present invention, the site 189, the site 234, the site 266 and the site 371 of the S-adenosylmethionine synthetase MAT are subjected to site-specific combinatorial mutation to obtain a series of mutants with improved enzyme activity, the I189V / V266H improvement is the most obvious, and the specific enzyme activity is 3.5833 + / -0.1373 U.mg <-1 >, which is improved by 1353.08% compared to the parent WT; and for a whole-cell catalysis system, the yield of S-ademetionine synthesized by the mutant I189V / V266H / Q234N / N371R reaches 1751.01 mg / L, and the increase is most obvious. The enzyme mutant provided by the invention has a wide application prospect in the production of S-adenosylmethionine and the construction of genetic engineering microorganisms of S-adenosylmethionine.

The invention relates to the technical field of gene engineering, in particular to a glycosyl transferase mutant, a sucrose synthase mutant and application of the glycosyl transferase mutant and the sucrose synthase mutant in synthesis of rebaudioside M. Wild glycosyl transferase and wild sucrose synthase are modified according to a directed evolution theory to obtain a glycosyl transferase mutant and a sucrose synthase mutant. Rebaudioside M is obtained by catalyzing rebaudioside D through a glycosyl transferase mutant and sucrose synthase mutant co-expression strain, and the catalytic efficiency is superior to that of a wild enzyme. The rebaudioside M is obtained by completely catalyzing 110 g / L of rebaudioside D through the co-expression strain of the glycosyl transferase mutant and the sucrose synthase mutant, only 20 h is needed, the conversion rate reaches 97%, the yield of the rebaudioside M reaches 122 g / L, the raw material conversion rate and the yield of the rebaudioside M are effectively increased, and meanwhile the synthesis time is shortened.

The invention relates to a series of GDP-fucose synthase polypeptides which can be applied to synthesis of GDP-fucose which is a precursor of human milk oligosaccharides (HMOs), and belongs to the technical field of enzyme engineering. In certain embodiments, the polypeptide capable of catalyzing the conversion of GDP-4-keto-6-deoxymannose into GDP-fucose comprises a polypeptide of which the amino acid sequence is at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of SEQ ID NO: 1 and / or the amino acid sequence of the polypeptide M1-14. The invention provides a series of polypeptides for synthesizing GDP-fucose.

The invention provides a method for producing borneol, the method comprising culturing the transgenic microorganism in a culture medium, and the microorganism comprises or expresses borneol diphosphate synthetase and phosphatase.

The invention relates to the field of bioengineering, in particular to an enzyme composition, a product and application of the enzyme composition in preparation of Ectoine. The invention provides an enzyme composition. The enzyme composition comprises a mutant of 2-hydroxy-4-aminobutyric acid oxidase, a mutant of keto transaminase, a mutant of hydroperoxide hydrolase, a mutant of Ectoine synthetase, a mutant of pyruvic acid-CoA oxidase and lactobacillus helveticus lactic dehydrogenase. According to the method disclosed by the invention, a plurality of mutant enzymes are utilized for combined transformation for the first time, and cheap 2-hydroxy-4-aminobutyric acid is taken as a starting raw material to quickly and efficiently produce Ectoine; the enzyme method has the characteristics of easiness in control, short conversion time, few generated impurities and the like, so that the method has unique advantages in the aspects of low-cost and large-scale mass production of Ectoin.

The invention discloses a yarrowia lipolytica engineering strain with high yield of resveratrol as well as construction and application of the yarrowia lipolytica engineering strain, and belongs to the technical field of synthetic biology and metabolic engineering. The invention provides a yarrowia lipolytica engineering strain. A 4-coumaric acid-CoA ligase gene, a multi-copy resveratrol synthase gene, a transketolase 1 gene, a fusion gene of a histidine phosphate aminotransferase gene and a tyrosine ammonia lyase gene, a glycolytic pathway hexokinase gene, a 6-phosphofructokinase-1 gene and a phosphoglycerate kinase gene are integrated, so that the recombinant protein is obtained. According to the present invention, by using glucose as the substrate, the high yield of resveratrol can be achieved by using glucose as the substrate, the resveratrol yield during shake-flask culture fermentation can achieve 3.157 g / L, the resveratrol yield can be increased to 30.7 g / L during 5L-scale fed-batch fermentation, and the resveratrol yield can be increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L; the maximum value is reported by the yarrowia lipolytica system at present.

The invention provides application of acidophilous glycosyltransferase in salidroside production, and belongs to the technical field of biological engineering. The problem of producing salidroside under the acidic condition is solved. Comprising an application of acidophilous glycosyl transferase with an amino acid sequence as shown in SEQ ID NO.1 in salidroside production under an acidic condition and an acidophilous escherichia coli engineering strain for producing salidroside. The escherichia coli engineering strain overexpresses a mutant 3-deoxy-D-arabinoheptulose-7-phosphate (DAHP) synthetase gene aroGfbr, a cyclohexadiene dehydrogenase gene tyrC, a glucose phosphate mutant enzyme gene pgm and a UDP-glucose pyrophosphorylase galU, overexpresses a phenylpyruvate decarboxylase gene ARO10 derived from saccharomyces cerevisiae, and can be used for producing a mutant 3-deoxy-D-arabinoheptulose-7-phosphate mutant enzyme. The kit comprises an ethanol dehydrogenase gene ADH6 and a glycosyl transferase gene LrUGT85AF8. The method is mainly used for producing salidroside under an acidic condition.

The application discloses a Weizmannia coagulans with fat-reducing and weight-reducing functions, and belongs to the technical field of microorganisms. The Weizmannia coagulans screened by the application can inhibit the content and activity of fat synthesis enzyme on the basis of reducing blood fat and weight, can relieve liver damage caused by high-fat diet, has excellent acid tolerance, bile salt tolerance and high fat hydrolysis capacity, and is a multifunctional Weizmannia coagulans, and has a wide application prospect in the preparation of fat-reducing, weight-reducing and weight-losing products.

The invention discloses construction and application of a saccharomyces cerevisiae engineering strain for synthesizing vanillin by taking ferulic acid as a substrate, and belongs to the technical field of genetic engineering. The invention provides a recombinant saccharomyces cerevisiae, which is characterized in that a series of genes for producing vanillic acid and vanillyl alcohol from vanillin through oxidation and reduction are knocked out, and feruloyl coenzyme A synthetase FCS and p-hydroxycinnamyl coenzyme A hydratase ECH which are obtained by screening from a metagene library in a West Sichuan hot spring extreme environment are overexpressed. According to the engineering strain VAN002 with high yield of vanillin, the accumulation of vanillin reaches 16.8 g / L, and the molar conversion rate of catalyzing ferulic acid to synthesize vanillin reaches 93%, which is the highest level reported by synthesizing vanillin by saccharomyces cerevisiae at present.

To identify lipid metabolites that control Th17 cell differentiation and lipid biosynthetic enzymes involved therein, and based on that, to provide a screening method for a therapeutic agent for diseases caused by excessive Th17 cell responses.SOLUTION: Provided is 1-oleoyl-lysophosphatidylethanolamine (LPE(1-18:1)) as a lipid metabolic product that controls Th17 cell differentiation. Since LPE(1-18:1) binds to RORγt with high specificity and promotes differentiation into Th17 cells, a candidate compound for a therapeutic agent for diseases caused by excessive Th17 cell responses can be obtained by selecting a compound that inhibits the binding of LPE(1-18:1) to RORγt. Also provided are five lipid metabolic enzymes (Gpam, Gpat3, Lplat1, Pla2g12a, and Scd2) that are responsible for the biosynthesis of LPE(1-18:1) and control Th17 cell differentiation. By selecting a compound that inhibits the activity or expression of these lipid metabolic enzymes, a candidate compound for a therapeutic agent for diseases caused by excessive Th17 cell responses can be obtained.SELECTED DRAWING: None

The invention discloses cloning and functional identification of a liverwort flavone synthetase gene, and belongs to the technical field of gene engineering. According to the invention, one flavone synthase I (flavones synthase I, FNS I) is obtained through screening, and is named as MpFNS I; a prokaryotic expression system is utilized, an MpFNS I recombinant protein is obtained through purification, and the result of in-vitro enzyme activity identification shows that an MpFNS I enzyme substrate is relatively wide in selectivity, has a flavone synthetase function and also has a part of flavonol synthetase function. The liverwort flavonoid has a significant difference on affinity and catalytic efficiency of different substrates, the in-vivo function of the liverwort flavonoid is verified by utilizing heterologous expression of arabidopsis thaliana, experimental evidence is provided for comprehensively analyzing a biosynthetic pathway of the liverwort flavonoid, and meanwhile, a research target is provided for deeply researching a mechanism for synthesizing and responding to UV-B radiation of the liverwort flavonoid.

The invention discloses a method for gene modification of aureobasidium pullulans, which belongs to the technical field of gene engineering, and comprises the following steps: determining a malic acid synthase gene, obtaining a malic acid synthase gene segment, searching a gene sequence of the malic acid synthase in the disclosed aureobasidium pullulans as a template on NCBI (National Center of Biotechnology Information), and carrying out gene modification on the malic acid synthase gene segment to obtain a malic acid synthase gene segment; the method comprises the following steps: respectively designing an upstream primer and a downstream primer, taking extracted genome DNA of aureobasidium pullulans M1 as a template, respectively carrying out PCR amplification and recombinant plasmid pUC57-PMA-ms construction and linearization treatment on the upstream primer and the downstream primer, and introducing the plasmid into the aureobasidium pullulans M1 to obtain an aureobasidium pullulans recombinant strain; according to the method, the polymalic acid synthetase gene (PMA-ms) on an aureobasidium pullulans genome is knocked out from aureobasidium pullulans, the yield of pullulan in the obtained recombinant plasmid M1-2 is about 39.11 g / L, and compared with an original strain M1, the yield of M1-2 is increased by 9.79 g / L, and the practical application prospect of the method is improved.

The invention discloses an artemisia apiacea transcription factor AaMYC3 with an amino acid sequence as shown in SEQ ID NO: 2 and application of the artemisia apiacea transcription factor AaMYC3. The Artemisia apiacea transcription factor AaMYC3 not only can be combined with and activate promoters of artemisinin biosynthesis key enzyme genes CYP71AV1 and ALDH1, but also can be combined with and activate a promoter of an Artemisia apiacea glandular hair development key gene AaHD1, and can act together with AabHLH1 and AabHLH113 to activate ADS and DBR2 gene promoters. Biosynthesis of artemisinin is promoted by positively regulating and controlling artemisinin biosynthase genes and glandular hair body density, and the method is of great significance to cultivation of new varieties with high artemisinin content and large-scale production.

Disclosed in the present invention are a sucrose synthase mutant and a method for preparing RD using same. The sucrose synthase mutant is based on the amino acid sequence shown in SEQ ID NO: 2 and contains a mutation in at least one of the following sites: D296Q, R636Q, G514L, H531L, R567T, D569Q, E610N, A642N, E663N, L667T and T742R. By means of site-directed mutagenesis of sucrose synthase, the enzyme activity of the sucrose synthase mutant is improved compared with that of the wild type. Also, some mutants can still maintain a high conversion rate in a wide pH range.

The application discloses a pyrano pyridine compound, a preparation method, a pharmaceutical composition and application thereof. The application provides a compound as shown in formula (I), a pharmaceutically acceptable salt or a stereoisomer thereof, the compounds have a strong inhibitory effect on aldosterone synthase, have little influence on cortisol synthase, have high selectivity and high safety, and can be used for preventing and / or treating various diseases related to aldosterone.

The invention discloses an amino acid sequence and application of a nucleotide sequence in synthesis of anisodine, and relates to the technical field of biology, the amino acid sequence is shown as SEQ ID NO.2, or the sequence shown as SEQ ID NO.2 is subjected to substitution, mutation or deletion of a plurality of amino acid residues, and the amino acid sequence with the same coding function is used for coding anisodine synthetase. The enzyme is used for catalytic synthesis of anisodine. The nucleotide sequence is as shown in SEQ ID NO.1, or the sequence as shown in SEQ ID NO.1 is substituted, mutated or deleted by a plurality of nucleotides, and the nucleotide sequence with the same coding function is used for coding the anisodine synthetase. An amino acid sequence for synthesizing the anisodine synthetase is found for the first time, the anisodine synthesis function of the anisodine synthetase is verified through a tobacco transient expression experiment, and the function of the anisodine synthetase is further verified through prokaryotic expression of in-vitro enzyme activity.

The invention belongs to the technical field of biology, and particularly relates to a genetically engineered bacterium capable of reducing by-products and improving yield of heparin precursors as well as a construction method and application of the genetically engineered bacterium. According to the technical scheme, acetic acid synthesis pathway genes ackA and poxB and lactic acid synthesis pathway genes ldhA are knocked out, and synthesis of acetic acid and lactic acid is blocked, so that more carbon metabolic flows enter heparin precursor synthesis pathways, the yield and conversion rate of heparin precursors are increased, and / or by-products are reduced. According to the invention, heparin precursor synthetases KfiA (UDP-GlcNAc glycosyl transferase), KfiC (UDP-GlcA glycosyl transferase) and KfiD (UDP-glucose dehydrogenase) are further enhanced, the expression of UDP-GlcA generated by UDP-Glc is catalyzed, and the yield and the conversion rate of the heparin precursor can be further improved and / or byproducts can be further reduced.

The invention belongs to the technical field of bioengineering, and relates to an ArsL mutant and an application of the ArsL mutant in synthesis of Arsinothricin. The ArsL mutant is a mutant obtained by mutating the 428th cysteine C of the amino acid sequence of wild AST synthetase ArsL into alanine A, and the amino acid sequence of the ArsL mutant is as shown in SEQ ID No.2. The invention also discloses a preparation method of the ArsL mutant. The invention also further provides an application of the ArsL mutant in catalysis of sodium methyl arsenate and S-adenosylmethionine to synthesize Arsinothricin. The invention further provides an application of the ArsL mutant in catalysis of sodium methyl arsenate and S-adenosylmethionine to synthesize Arsinothricin. The ArsL mutant disclosed by the invention has higher catalytic activity and thermal stability, the process of converting high-concentration MMA into AST is realized by utilizing the ArsL mutant, and the catalytic activity of the ArsLC428A mutant is obviously improved compared with that of wild type AST synthetase. In addition, compared with an organic synthesis path, a semi-organic enzyme synthesis path and a natural biological synthesis path, the yield of the synthesis path provided by the invention is obviously improved, and the ArsL mutant can be used as a potential industrial application enzyme and has a wide application prospect.

The invention relates to the technical field of biology, in particular to a mutant of difunctional glutathione synthetase and application of the mutant. The amino acid sequence of the mutant of the difunctional glutathione synthetase provided by the invention is obtained by performing site-directed mutagenesis on at least one of the 597th site, the 607th site, the 609th site, the 610th site, the 728th site and the 733th site of the amino acid sequence of the wild type difunctional glutathione synthetase, and the amino acid sequence of the wild type difunctional glutathione synthetase is shown as SEQ ID NO: 1. The mutant of the bifunctional glutathione synthetase provided by the invention has relatively high enzyme activity and enzyme stability, is more suitable for industrial production and is suitable for preparing glutathione, and the efficiency of preparing glutathione is improved.

This application relates to a method for producing a protein containing an unnatural amino acid (UAA), the method comprising culturing a host cell, wherein the host cell is a eukaryotic cell, together with: a nucleotide sequence encoding a first recoding tRNA or a first recoding tRNA, wherein the first recoding tRNA comprises an anticodon complementary to a first codon, and wherein the first codon is a rare codon; and a nucleotide sequence encoding a first aminoacyl-tRNA synthetase or a first aminoacyl-tRNA synthetase, wherein the first aminoacyl-tRNA synthetase is capable of charging the first recoding tRNA with the first unnatural amino acid.

The invention relates to the technical field of plant genetic engineering, in particular to a medicago sativa MsSNAT1 gene and application of the medicago sativa MsSNAT1 gene in regulation and control of plant salt tolerance. The nucleotide sequence of the medicago sativa MsSNAT1 gene is as shown in SEQ ID NO. 1. Experimental results show that overexpression of the MsSNAT1 gene enhances the stress resistance of medicago sativa. The overexpression of the MsSNAT1 gene promotes the increase of the content of endogenous melatonin and increases the expression of the melatonin synthetase gene under salt stress. Compared with a wild type, the antioxidant enzyme activity and osmoregulation substance increase trend of the transgenic line under salt and drought stress is more obvious, the MDA content is lower, and the negative influence is smaller. In addition, the content of endogenous melatonin in a transgenic line is increased, and the endogenous melatonin and exogenous ABA act together, so that the oxidation resistance and osmotic regulation capacity of the plant are enhanced, and the stress resistance of the plant is remarkably improved.

The invention discloses yarrowia lipolytica with high yield of beta-carotene as well as a construction method and application of the yarrowia lipolytica. According to the yarrowia lipolytica engineering bacterium, geranyl diphosphate synthase gene xdGPS, phytoene dehydrogenase gene CarB, phytoene cyclization / phytoene synthesis bifunctional enzyme mutant gene GarRPY27R, acetyl-coenzyme A synthetase mutant gene ACSL641P and pyruvate ferredoxin oxidoreductase gene nifJ are integrated and expressed on a chromosome, and the yarrowia lipolytica engineering bacterium is obtained. Meanwhile, a 3-hydroxy-3-methylglutaryl CoA reductase gene HMGR (3-hydroxy-3-methylglutaryl CoA reductase) derived from the yarrowia lipolytica strain is subjected to overexpression; the yield of the beta-carotene is increased to 13.186 g / L, and the production efficiency is 0.14 g / L / h.

The present invention relates to peptidomimetics of a short peptide from leucyl-tRNA synthetase, compositions comprising one or more of said peptidomimetics and their use for the treatment of syndromes caused by mutations of mt-tRNA (mitochondrial transfer RNA) genes, and medical treatments of said syndromes comprising the administration of said one or more peptidomimetics or compositions comprising the same.

A recombinant microbial cell is capable of producing at least one compound having structural Formula II from a carbon source:In Formula II, n=2-3; R1=H or COCH3 or CH2CH2COX with X=OH or O—; R2=CH3 or CH2CH2COX with X=OH or O— where the cell comprises a genetic modification to increase activity relative to its wild-type cell of E4 where E4 is a desferrioxamine or bisucaberin synthetase (EC 6.3.-.-) (E4i) capable of converting N5-aminopentyl-N-(hydroxy)-succinamic acid to desferrioxamine B or H or at least one other linear desferrioxamine or bisucaberin according to Formula II.