423 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 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 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 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 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 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.

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 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 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 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.

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.

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

A Levodopa-selective sensor is provided. A continuous levodopa monitor is also provided. The levodopa sensor includes a sensor area having a working electrode with a levodopa-selective chemistry configured for at least partial implantation in a host. The sensor also includes at least one membrane adjacent the levodopa-selective chemistry. The at least one membrane includes an enzyme domain that at least one enzyme selected from a tyrosinase; a mutated tyrosinase with specificity towards I-3,4-dihydroxyphenylalanine; a dihydroxyphenylalanine 4,5-dioxygenase; a mutated dihydroxyphenylalanine 4,5-dioxygenase with specificity towards I-3,4-dihydroxyphenylalanine; and combinations thereof; a synthase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; a mutated synthase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; a 3,4-dihydroxyphenyl-acetaldehyde synthase; a mutated 3,4- dihydroxyphenyl-acetaldehyde synthase with specificity towards I-3,4-dihydroxyphenylalanine; a dioxygenase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; and a mutated dioxygenase enzyme with specificity towards I-3,4-dihydroxyphenylalanine.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a promoter of a pinus massoniana terpene synthase gene Pm TPS (-)-alpha-pinene and application of the promoter. According to the invention, a promoter fragment of a Pm TPS (-)-alpha-pinene gene and three homeopathic action elements with jasmonic acid response on the promoter fragment are obtained from pinus massoniana. A complete promoter fragment of the Pm TPS (-)-alpha-pinene gene of the pinus massoniana is transformed into arabidopsis thaliana by means of agrobacterium mediation, and the promoter can drive expression of a GUS gene in leaves of the arabidopsis thaliana or nicotiana benthamiana. GUS staining and GUS enzyme activity of transgenic Bensi tobacco leaves are reduced along with deletion of jasmonic acid response cis-acting elements on the promoter. Furthermore, the yield of pine resin terpenoids can be increased by utilizing the promoter and the homeopathic element thereof.

The invention belongs to the technical field of facility horticulture postharvest treatment, and particularly relates to a tomato postharvest quality regulation and control light supplementing system based on white light-metabolism key enzyme coupling activation and an application method thereof, and the tomato postharvest quality regulation and control light supplementing system comprises an enzyme activity targeting white light module, an enzyme activity feedback module and a fruit bearing regulation and control module. According to the postharvest tomato quality regulation and control light supplementing system based on white light-metabolic key enzyme coupling activation, a white light spectrum segmentation-key enzyme activity dynamic matching coupling mechanism is innovatively designed by combining the regulation and control rule of white light on the activity of key enzymes such as carotenoid synthetase and phenylpropane metabolic enzyme of tomato fruits; through an adjustable broad-spectrum white light source and an enzyme activity feedback module, directional accumulation of beneficial metabolites such as carotenoids and flavonoid compounds and efficient degradation of alkaloids are realized.

The invention relates to the fields of enzyme engineering and microbial fermentation production, in particular to an N-acetylglutamate synthetase mutant for producing N-acetyl blue and application of the N-acetylglutamate synthetase mutant. According to the invention, the N-acetylglutamate synthetase mutant EcargAV323A / S435A is constructed by researching the structure of N-acetylglutamate synthetase ArgA (EcargA) from Escherichia coli and mutating the 323rd site and the 435th site of EcargA protein in combination with a directed evolution technology, so that the N-acetylglutamate synthetase mutant EcargAV323A / S435A is constructed. When the engineering strain of the N-acetylglutamate synthetase mutant is used for producing the N-acetylobservation blue through fermentation, the yield of the N-acetylobservation blue can be remarkably improved, the generation of a by-product observation blue is reduced, the fermentation process is simplified, and the production cost is reduced. The N-acetylglutamate synthetase mutant and the engineering strain provided by the invention have good industrial application value.

The invention discloses a glycolaldehyde synthetase mutant and application of the glycolaldehyde synthetase mutant in improving the thermal stability of glycolaldehyde synthetase, and belongs to the technical field of biological catalysis application. According to the glycolaldehyde synthetase mutant disclosed by the invention, on the basis of an amino acid sequence as shown in SEQ ID NO.1, the glycolaldehyde synthetase mutant has the following mutations: A381P / E509F, A381P / K290P, A381P, S61A, A381P / K290P / E509F, A381P / C49A, A381P / T100V, A381P / S61A or A381P / K290P / S61A. The mutant protein can catalyze formaldehyde to prepare hydroxyacetaldehyde (glycolaldehyde). In addition, the mutant protein can also improve the catalytic efficiency from formaldehyde to glycolaldehyde, and due to the improvement of the thermal stability, continuous catalysis of industrial enzyme in actual reaction is facilitated, so that the mutant has a good industrial application prospect.

Provided herein are novel materials and methods for site-specific incorporation of phosphotyrosines into proteins. The novel methods of the invention encompass the use of a novel aminoacyl tRNA synthetase capable of charging compatible tRNAs with a phosphotyrosine precursor. The phosphotyrosine precursor is then incorporated, site-specifically, into a protein at sites where phosphotyrosine residues are desired. The phosphotyrosine precursors are subsequently treated to convert them into phosphotyrosine residues, yielding proteins with phosphotyrosines at selected sites. The scope of the invention encompasses novel aminoacyl tRNA synthetases, novel phosphotyrosine precursors, and methods of using these materials to create site-specific phosphorylated tyrosine residues in a protein.

This disclosure relates to modified tryptophan synthase and more particularly to modified beta-subunits of tryptophan synthase. The disclosure further relates to cells expressing such modified subunits and methods of producing non-canonical amino acids.

The invention discloses recombinant saccharomyces cerevisiae, a 11-seed oil fermentation product as well as a preparation method and application of the 11-seed oil fermentation product, and belongs to the technical field of synthetic biology and fermentation product preparation. According to the recombinant saccharomyces cerevisiae disclosed by the invention, a caffeic acid coenzyme A ligase 4CL coding gene, a chalcone isomerase CHI coding gene, a flavanone-3-hydroxylase F3H coding gene and a chalcone synthase CHS mutant coding gene are over-expressed, and the chalcone synthase CHS mutant is preferably Y69H / H71Y / Q161K. The preparation method of the 11-seed oil fermentation product comprises the following steps: fermenting a recombinant saccharomyces cerevisiae fermentation solution in the presence of a sophora flower bud extract and caffeic acid to obtain a first fermentation solution; fermenting the lactic acid bacteria fermentation liquor in the presence of 11 seed oil to obtain second fermentation liquor; and mixing the first fermentation liquid and the second fermentation liquid, continuing fermentation, and respectively collecting an oil phase and a water phase after fermentation is finished, so as to obtain a fermentation product. The product provided by the invention has multivitamin antioxidant activity.

The invention discloses a sucrose synthase mutant and a method for preparing RD (Reduced Disorder) by using the sucrose synthase mutant. The sucrose synthase mutant is obtained by mutation of at least one of the following sites on the basis of an amino acid sequence as shown in SEQ ID NO: 2: D296Q, R636Q, G514L, H531L, R567T, D569Q, E610N, A642N, E663N, L667T and T742R; site-directed mutagenesis is carried out on sucrose synthetase, so that the enzyme activity of the sucrose synthetase mutant is improved compared with that of a wild type, and meanwhile, a part of the mutant still maintains a relatively high conversion rate in a relatively wide pH range.

The invention discloses a fusarium venetianum engineering strain as well as a construction method and application thereof, relates to the technical field of gene engineering, and aims to solve the problem of low fungal hypha protein yield, chitin synthetase FvChs4 gene of fusarium venetianum is deleted, the classification name of the fusarium venetianum engineering strain is fusarium venetianum, and the fusarium venetianum engineering strain has the characteristics that the fusarium venetianum engineering strain has a nucleotide sequence shown in SEQ ID NO: 1 and a nucleotide sequence shown in SEQ ID NO: 2 and a nucleotide sequence shown in SEQ ID NO: 3. The fusarium venanii engineering strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) on May 27, 2025, and the preservation number is CGMCC No. 42015. Through targeted knockout of the FvChs4 gene, consumption of a carbon source by cell wall synthesis is reduced, and directional enrichment of the carbon source to protein synthesis is realized, so that the yield of hypha protein is increased.

The present invention relates to the field of biotechnology, and more particularly to a highly active S-sulfo-L-cysteine ​​synthetase mutant and its applications. The present invention screened and identified a preferred SSC03 enzyme through cluster analysis and phylogenetic analysis, and further engineered mutations thereof. The resulting SSC03-S256PΔCT achieved a 24-hour conversion rate of 56% and produced 84 mM of S-sulfo-L-cysteine, demonstrating its applicability to further purification process development.