106 results about "Dehydratase" patented technology

The invention belongs to the field of biology, and particularly relates to a fusion protein for promoting synthesis of 1, 3-propylene glycol and application of the fusion protein. The fusion protein NADPH (Nicotinamide Adenine Dinucleotide Phosphate) dependent alcohol dehydrogenase fusion protein or NADH (Nicotinamide Adenine Dinucleotide Hormone) dependent alcohol dehydrogenase fusion protein disclosed by the invention can be packaged into diol dehydratase and can promote the synthesis of 1, 3-propylene glycol. Experiments prove that the bacillus subtilis has an effect of improving the fermentation performance of the strain.

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention relates to a Kluyveromyces cerevisiae gene modified cell containing GDP-mannose dehydratase polypeptide, which can be applied to synthesis of human milk oligosaccharide, and belongs to the field of synthetic biology. According to the technical scheme, the gene modified cell comprises a recombinant nucleic acid sequence for coding a polypeptide with GDP-mannose dehydratase polypeptide (GDP-mannose dehydratase polypeptide) activity. The polypeptide capable of catalyzing the conversion of GDP-mannose into GDP-4-keto-6-deoxymannose comprises a polypeptide of which the amino acid sequence is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2 and / or the amino acid sequence of polypeptides M1-M13, wherein the amino acid sequence of the polypeptide is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to the amino acid sequence of the polypeptides M1-M13. The invention provides a kluyveromyces kluyveromyces genetically modified cell with fucosyllactose synthesis activity.

The invention discloses a production method of fucosylated oligosaccharide. The core of the method is as follows: the GDP-D-rhamnose is converted into the GDP-L-fucose by using the GDP-D-rhamnose-3, 5-epimerase; the GDP-D-rhamnose is obtained by converting the GDP-D-mannose-4, 6-dehydratase and the GDP-4-keto-6-deoxy D-mannose reductase by taking the GDP-D-mannose as a substrate, and the GDP-D-rhamnose is obtained by taking the GDP-D-mannose as a substrate. The invention relates to a method for producing a fucosylated oligosaccharide, which is characterized in that a GDP-D-mannose-4, 6-dehydratase, a GDP-4-keto-6-deoxyD-mannose reductase, a GDP-D-rhamnose-3, 5-epimerase and a fucosyltransferase are expressed in a genetically modified host cell, and the fucosylated oligosaccharide can be produced by the way mentioned above.

The invention discloses recombinant yarrowia lipolytica with high yield of 3-fucosyllactose as well as a construction method and application of the recombinant yarrowia lipolytica. According to the invention, a reinforced lactose transporter lac12 gene, a reinforced GDP-mannose-4, 6-dehydratase gmd gene, a reinforced GDP-L-fucose synthase gmer gene and a 3-fucosyltransferase SUMO-fut3Bc gene with a reinforced SUMO tag are subjected to integrated expression on a recombinant yarrowia lipolytica chromosome, and a 6-phosphogluconolactonase pgl gene is subjected to overexpression, so that a recombinant yarrowia lipolytica strain is obtained. The constructed recombinant yarrowia lipolytica can maintain the genetic stability of a target gene, the yield and production efficiency of 3-fucosyllactose are remarkably improved, and the requirements of industrial production can be met.

The invention discloses a Zymomonas mobilis recombinant strain for producing D-pantothenic acid as well as a preparation method and application of the Zymomonas mobilis recombinant strain. A hydroxymethyl transferase gene panB, a ketopantoic acid reductase gene panE and a pantothenic acid synthase gene panC are integrated in a genome of the recombinant strain; an expression plasmid containing an acetolactate synthase gene Bsals, a keto acid reductoisomerase gene ilvC and a dihydroxy acid dehydratase gene ilvD is also transferred into the recombinant strain; wherein the nucleotide sequences of the gene Bsals, the gene ilvC, the gene ilvD, the gene panB, the gene panE and the gene panC are shown as SEQ ID NO. 1 to SEQ ID NO. 6 in sequence. The zymomonas mobilis ZM4 is systematically modified through a genetic engineering means, and an optimal enzyme combination adaptive to the zymomonas mobilis is defined by screening hydroxymethyltransferase, pantothenic acid synthase and ketopantoic acid reductase from different sources, so that a core foundation is laid for efficient operation of a D-pantothenic acid synthesis route, and the method has a wide application prospect. The technical bottlenecks of single metabolic pathway and low synthesis efficiency of the traditional strain are broken through.

Provided herein are host cells capable of producing a human milk oligosaccharide (HMO), such as yeast cells that include one or more heterologous nucleic acids encoding one or more enzymes of the HMO biosynthetic pathway, such as a fucosyltransferase, GDP-mannose dehydratase, lactose permease, and / or fucose synthase. Also provided are fermentation compositions including the disclosed host cells, as well as related methods of producing and recovering HMOs generated by the host cells.

The invention provides a series of GDP-mannose dehydratase polypeptides which can be applied to synthesis of GDP-fucose, and belongs to the technical field of enzyme engineering. The GDP-mannose dehydratase polypeptide comprises a polypeptide of which the amino acid sequence is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to that of a polypeptide shown in SEQ ID NO: 1, or a polypeptide of which the amino acid sequence is at least 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical to that of a polypeptide M1-13. The GDP-mannose dehydratase polypeptide disclosed by the invention has relatively high catalytic activity.

The invention discloses a recombinant bacterium capable of increasing the yield of alpha-bisabolol as well as a preparation method and application of the recombinant bacterium, and belongs to the technical field of microorganisms. The invention aims to improve the yield of alpha-bisabolol and enhance the tolerance of a host to an organic solvent. The invention provides a recombinant bacterium for improving the yield of alpha-bisabolol. Escherichia coli is used as a starting strain; the method comprises the following steps of: overexpressing an acetyl CoA acyltransferase / HMG-CoA reductase mvaE gene, an HMG-CoA synthetase mvaS gene, a 2-methyl citrate dehydratase prpD gene, a mevalonate kinase ERG12 gene, a mevalonate 5-phosphate kinase ERG8 gene, a mevalonate 5-diphosphate decarboxylase ERG19 gene and an isopentenyl diphosphate isomerase idi gene, so as to obtain a recombinant vector; the gene is obtained from an alpha-bisabolol synthase gene of artichoke, a farnesyl diphosphate synthase ispA gene and an alpha-bisabolol synthase CcBOS gene of artichoke. The industrial process of synthesizing alpha-bisabolol by a biological method is promoted.

This invention relates to the fields of microbial genetic engineering and fermentation biomanufacturing technology, specifically disclosing a recombinant Corynebacterium glutamicum with high 5-aminolevulinic acid (ALA) production and its applications. Addressing the problems of existing strains such as crude enzyme activity regulation, inhibited cell growth, the need for external inhibitors, and low yield, this invention uses Corynebacterium glutamicum as a host and removes the gdhA gene without scarring to reduce precursor competitive metabolism. Site-directed mutations of the endogenous hemB gene (S288A, D128E, R231K) are performed to moderately reduce dehydratase activity. A high-expression plasmid pXMJ19-hemA containing the hemA gene from Rhodopsinus capsulatum is constructed and introduced into the modified strain to obtain a recombinant strain with optimized triple metabolism. This strain reduces product degradation without the need for external dehydratase inhibitors, does not affect cell growth, and achieves a yield up to 4.30 times higher than the control after 42 hours of fermentation, with ALA accumulation exceeding 8.9 g / L. The strain exhibits stable characteristics, the process is simple, and it has good industrial application value.

The present invention discloses a method for synthesizing hydroxytyrosol by multi-enzyme cascade, comprising: 1) preparing a hydroxytyrosol product by using 3,4-dihydroxybenzaldehyde, L-threonine, sodium formate, NAD + As raw material, L-threonine transaldolase, alcohol dehydrogenase, and formate dehydrogenase are used to catalyze the synthesis of L-threo-3-(3,4-dihydroxyphenyl)serine; 2) L-threo-3-(3,4-dihydroxyphenyl)serine is used as substrate, glucose, NAD + Hydroxytyrosol is synthesized through a cascade catalysis of phenylserine dehydratase, α-ketoacid decarboxylase, aldehyde reductase, and glucose dehydrogenase. The synthesis method provided by the present invention achieves a hydroxytyrosol yield of >99% and a space-time yield of 0.88 g / L / h, currently the highest, showing great potential for industrial application and suitable for further promotion and application.

The invention discloses construction and application of genetically engineered bacteria for high-yield rose honey sweet essence. The genetically engineered bacterium is obtained by overexpressing tobacco pre-benzoic acid dehydratase NtPHA2 and an ARO80 mutant ARO80S706T / L877Q / E910D by virtue of saccharomyces cerevisiae. According to the invention, NtPHA2 and an ARO80 mutant ARO80S706T / L877Q / E910D are over-expressed in the saccharomyces cerevisiae, so that the biosynthesis yield of rose honey sweet aroma substances such as 2-phenethyl alcohol and the like is increased, and the rose honey sweet essence can be produced in batches by utilizing the engineering strain.

The invention discloses a preparation method and application of fucose and an intermediate thereof. The method for synthesizing the GDP-fucose comprises the following steps: taking guanosine diphosphate and cane sugar as substrates, and carrying out catalytic reaction on cane sugar synthase, CDP-tetrose 2-epimerase, GDP-mannose dehydratase and isoreductase to generate the GDP-fucose. The method for synthesizing the fucosyllactose comprises the following steps: taking GDP-fucose and pNP-lactose as initial substrates, and obtaining the fucosyllactose under the catalytic action of Hp13FucT2 enzyme. According to the method for preparing the L-fucose through the enzyme method, fucosyllactose is used as a substrate, and the L-fucose is generated through a fucosidase catalytic reaction. The adopted large intestine expression system has the characteristics of high expression efficiency, large expression quantity, low cost, easiness in operation and the like, and the expressed fucosidase has the advantages of high purity, high yield and the like.

The invention discloses an application of a soybean 2-hydroxyisoflavanone dehydratase gene in improving the growth of intercropping corn. The gene HID1 can promote soybean to synthesize isoflavone, recruit bacillus and promote mycorrhizal fungi infection, so that the growth of intercropped corn is promoted, and the gene HID1 has important significance in illuminating the biological function of the GmHID1 gene in leguminous crop intercropping and regulating and controlling the synergistic growth promotion of plants, mycorrhizal fungi and bacillus in intercropping.

The invention relates to a recombinant halomonas, which comprises an ilvC (acetohydroxy acid isoreductase, AHAIR) gene, an ilvD (dihydroxy acid dehydratase, DHAD) gene and an optional ilvE (transaminase, TA) gene, wherein the ilvC (acetohydroxy acid isoreductase, AHAIR) gene and the ilvD (dihydroxy acid dehydratase, DHAD) gene are externally introduced. According to the recombinant halomonas disclosed by the invention, the L-leucine and / or the L-isoleucine can be efficiently produced without sterilization, in an energy-saving and water-saving manner through open type fermentation, and the industrial production of the recombinant halomonas is facilitated.

This invention belongs to the fields of fermentation and genetic engineering technology, and provides a high-yield hexanoic acid-producing engineered *Escherichia coli* strain, its construction method, and its applications. This engineered strain is *Escherichia coli*... E. coli w3110 was the starting strain, overexpressing the acetyl-CoA transferase gene. act and β-ketothiolase gene bktB Overexpression of trans-enoyl-CoA reductase gene ter 3-Hydroxybutyryl-CoA dehydrogenase gene hbd, and 3-hydroxybutyryl coenzyme A dehydratase gene crt Or, simultaneously not expressing the pyruvate formate lyase gene. pflB or lactate dehydrogenase gene ldhA This engineered bacteria can convert glucose into hexanoic acid, a high-value product, with a yield of 4.968 g / L after 18 hours of fermentation. It adopts aerobic fermentation, resulting in rapid cell growth, a short fermentation cycle, and a high acid production rate. The fermentation process is simple, easy to control, and has low production costs, which is conducive to its promotion and application in industrial production.

The invention provides genetically engineered saccharomycetes capable of simultaneously producing baicalein and baicalin as well as a construction method and application of the genetically engineered saccharomycetes. According to the invention, by weakening or deleting a glucose 5-phosphate dehydrogenase gene (ZWF1) and phenylpyruvate decarboxylase genes ARO10 and PDC5 in a saccharomyces cerevisiae genome and enhancing the expression of a pre-benzoic acid dehydratase gene Pha2, the metabolic flow of a shikimic acid pathway is increased, the aldrin pathway is blocked, and the yield of the shikimic acid pathway is increased. Therefore, the yield of L-phenylalanine serving as a starting material for synthesizing baicalein and baicalin is increased; meanwhile, exogenous genes PAL, 4CL, CHS, CHI, FNSII, F6H, CPR and F7GAT are integrated onto the genome by virtue of a gene recombination technology, so that baicalein and baicalin are produced from the beginning by taking glucose and glycerol as a mixed carbon source without adding precursor substances such as exogenous L-phenylalanine or malonyl CoA and the like.

A method of synthesizing(S)-3-hydroxy-γ-butyrolactone (HBL), the method including producing a trione using an enzymatic conversion of glucosone. The enzymatic conversion may include: mutating a domain of Aldos-2-ulose dehydratase (AUDH) to obtain a mutant AUDH; dehydrating glucosone using the mutant AUDH to obtain an intermediate, basidiopyrone P (BPP) (4,5-dihydroxy-2-(hydroxymethyl)-2H-pyran-5-one); and forming the trione from the BPP. The method may further include catalyzing the trione using a base to form an ester of 3,4-dihydroxybutyrate; and converting the ester of 3,4-dihydroxybutyrate into the HBL by adding an acid to the ester of 3,4-dihydroxybutyrate.

The disclosure relates to the field of specialty chemicals and methods for their synthesis. In embodiments, the disclosure provides viable bacterial cells which comprise heterologous dual 3-hydroxy-acyl-ACP dehydratase / isomerases, etc. The disclosure further provides monounsaturated fatty acid derivative molecules produced by the viable bacterial cells which are non-native to the bacterial cells. The disclosure further provides methods for the preparation and production of non-native monounsaturated fatty acid derivative molecules such as e.g., an ω3-monounsaturated fatty acid derivative, an ω5-monounsaturated fatty acid derivative, an ω9-monounsaturated fatty acid derivative, an ω11-monounsaturated fatty acid fatty acid derivative, etc.

Disclosed in the present invention is the use of a threonine dehydratase in the regulation of the growth of Didymella segeticola and as a bactericide target. The threonine dehydratase gene Td of the present invention is derived from Didymella segeticola, and a knockout mutant is obtained by means of constructing a knockout gene fragment targeting the Td gene and then introducing same into a D. segeticola protoplast. Experiments find that the Td gene plays a role in the growth, the in-vivo pyruvic acid content and the in-vivo ATP content of D. segeticola, and the sensitivity thereof to wuyiencin. By means of molecular docking analysis, molecular dynamics simulation and microscale thermophoresis experiments, it is verified that threonine dehydratase can be used as a bactericide target, and the bactericide wuyiencin is obtained by means of screening. The threonine dehydratase of the present invention can be used as a bactericide target and a key protein in the pathogenic mechanism of a disease, and has broad application prospects in the control of plant pathogenic fungal diseases.

The invention discloses a recombinant microorganism for producing alpha-ketoisovaleric acid as well as a preparation method and application of the recombinant microorganism. The recombinant microorganism for producing alpha-ketoisovaleric acid, disclosed by the invention, has the following characteristics: the recombinant microorganism contains a substance capable of expressing an acetolactate synthase gene and / or a substance capable of promoting the expression of the acetolactate synthase gene and / or a substance capable of improving the activity or content of acetolactate synthase; the composition contains a substance capable of expressing an acetohydroxy acid reductoisomerase gene and / or a substance capable of promoting the expression of the acetohydroxy acid reductoisomerase gene and / or a substance capable of improving the activity or content of acetohydroxy acid reductoisomerase; the composition contains a substance capable of expressing a dihydroxy acid dehydratase gene and / or a substance capable of promoting the expression of the dihydroxy acid dehydratase gene and / or a substance capable of improving the activity or content of the dihydroxy acid dehydratase. Experiments prove that the recombinant microorganism disclosed by the invention can be used for producing alpha-ketoisovaleric acid by utilizing glucose and has a very good application prospect.

The invention provides a yeast engineering strain for synthesizing melatonin as well as a construction method and application of the yeast engineering strain, and relates to the technical field of genetic engineering and microbial fermentation engineering. According to the invention, a novel yeast engineering bacterium for synthesizing melatonin is constructed, wherein the novel yeast engineering bacterium is used for overexpressing arylalkylamine N-acetyl transferase, 5-hydroxytryptophan decarboxylase, tryptophan hydroxylase, N-acetyl serotonin methyltransferase, 6-pyruvoyl tetrahydrobiopterin synthetase, ink pterin reductase, 4a-hydroxytetrahydrobiopterin dehydratase and dihydropterin reductase. Under the fermentation process that 1g / L tryptophan is externally added, the content of melatonin in a fermentation product obtained by the yeast engineering bacteria reaches 14.32 mg / L, and 17.48 mg / g DCW. Under the condition that 4g / L tryptophan is added, 5L fermentation tank culture is carried out, and the content of melatonin in a fermentation product obtained by the yeast engineering bacteria reaches 91.82 mg / L. The engineered yeast provided by the invention can significantly increase the yield of melatonin, is expected to realize industrial mass production of melatonin, and has good application value and commercial value.

Provided are a genetically engineered bacterium for high-yield production of D-pantothenic acid (D-PA), and a preparation method therefor and a use thereof. The D-PA production yield of the engineered bacterium is significantly improved by means of the following actions: overexpressing acetolactate synthases IlvB and IlvN for pyruvate diversion while deleting a transcription attenuation region and mutating a valine feedback inhibition site; overexpressing ketol-acid reductoisomerase IlvC and dihydroxy acid dehydratase IlvD, and altering the coenzyme preference of IlvC from NADPH to NADH to unlock the upstream pathway for D-PA synthesis; overexpressing PanB and overexpressing a serine-glycine transport system to promote the synthesis of 5,10-methylenetetrahydrofolate; at the same time, enhancing the expression of ketopantoic acid reductase PanE; and finally overexpressing pantothenate synthetase PanC and increasing the content of cofactor ATP required by PanC. By means of the integrated enhancement of the above multiple modules, the provided engineered bacterium achieves improved cell growth due to balanced cofactors and increased D-PA.

The invention belongs to the technical field of gene engineering, and discloses a method for synthesizing L-cysteine through whole-cell catalysis, recombinant Escherichia coli and application, and the recombinant Escherichia coli is constructed through the following method: knocking out sdaA and tdcG genes for coding L-serine dehydratase; mutant beta subunit genes are obtained to construct recombinant plasmids, and the recombinant plasmids efficiently overexpress mutant genes and genes trpA in strains to obtain recombinant escherichia coli. The method is mild in reaction condition, low in energy consumption, environmentally friendly and high in substrate utilization rate through an enzyme method, and the overall cost is obviously reduced. The catalytic system can efficiently react at 25-35 DEG C from neutral to alkalescence, high temperature and high pressure or toxic chemical reagents are not needed, and energy consumption and environmental pressure are remarkably reduced. Meanwhile, the reaction system is high in substrate conversion rate and few in side reaction, the utilization rate of raw materials is greatly increased, and the method has obvious economic advantages.

The invention provides a verticillium dahliae mutant strain as well as a construction method and application thereof, a homologous recombination technology is adopted to replace a cotylidone dehydratase encoding gene with a hygromycin gene, a genetic engineering strain capable of accumulating a large amount of cotylidone under conventional culture conditions is obtained, the strain is simple in culture process, stable in phenotype and high in yield, and the yield of cotylidone is increased. The method can be used for producing the cotylidone, and the fungus melanin is synthesized by using the cotylidone, so that the defects of time consumption, complicated steps and the like in the traditional fungus melanin extraction method are overcome.

This disclosure provides glucarate dehydratase enzymes useful for commercial scale production of 5-keto-4-deoxy glucaric acid (KDG) from glucaric acid. The produced KDG may be dehydrated to form furan dicarboxylic acid (FDCA). The disclosed enzymes ("dehydratase variants") are variants of Lactipantibacillus pentous dehydratase engineered to have improved thermal tolerance and resistance to inhibition by tartrate as compared with the parent enzyme.

The application discloses tobacco isopropyl malate dehydrase NtIPDS, a coding gene, an RNAi interference carrier and application. The application successfully clones a coding gene of isopropyl malate dehydrase in tobacco, and the nucleotide sequence is as follows: (1) the nucleotide sequence shown in SEQ ID NO. 1; (2) the nucleotide sequence shown in SEQ ID NO. 1 is substituted and / or deleted and / or added with one or more nucleotides and the nucleotide sequence of the same functional protein. Based on the existing tobacco genetic engineering progress and related plant phenotype observation and identification, the NtIPDS gene is deeply researched, and it is found that the NtIPDS gene is related to tobacco chloroplast development and acyl sugar metabolism. After the gene is silenced, the chlorosis plant growth is slow, the chloroplast development is found to be abnormal through transmission electron microscope observation, and meanwhile, it is found that the acyl sugar content of the silenced plant is significantly increased through metabolic detection analysis.

The invention provides a genetically engineered recombinant Klebsiella michiganensis having reduced or inactivated glucose-specific PTS transport system, glycerol kinase, glycerol dehydrogenase, alcohol dehydrogenase, acetolactate synthase, acetolactate decarboxylase, fumarate reductase and 3-phosphoglyceraldehyde dehydrogenase, having, preferably enhanced, NADPH-dependent alcohol dehydrogenase, diol dehydratase, 3-phosphoglycerate dehydrogenase and 3-phosphoglycerate phosphatase activities, methods of making said recombinant Klebsiella michiganensis, and uses and methods of producing 1,3-propanediol using said recombinant Klebsiella michiganensis.

The application relates to the technical field of microbial engineering, and particularly discloses a dihydroxy acid dehydratase mutant, a recombinant microorganism thereof and application. The dihydroxy acid dehydratase mutant takes the amino acid sequence of wild-type dihydroxy acid dehydratase as a reference sequence, and contains a mutation that the 237th alanine is substituted by lysine, arginine, histidine or proline. The fermentation bacteria with the mutant of the application have obviously improved L-valine production capacity, high yield and few by-products. The application provides a new high-efficiency production mode for fermentative production of L-valine.

A method for producing fucosylated oligosaccharides. The core of the method lies in converting GDP-D-rhamnose into GDP-L-fucose by using GDP-D-rhamnose-3,5-epimerase, wherein the GDP-D-rhamnose is obtained by means of converting GDP-D-mannose as a substrate via GDP-D-mannose-4,6-dehydratase and GDP-4-keto-6-deoxy-D-mannose reductase. The expression of GDP-D-mannose-4,6-dehydratase, GDP-4-keto-6-deoxy-D-mannose reductase, GDP-D-rhamnose-3,5-epimerase, and fucosyltransferase in genetically modified host cells enables the production of fucosylated oligosaccharides via the described pathway.