114 results about "Corynebacterium" patented technology

The invention discloses a corynebacterium glutamicum engineering strain produced by L-glutamine as well as a construction method and application of the corynebacterium glutamicum engineering strain. The corynebacterium glutamicum engineering strain is prepared by integrating a glutamine synthase encoding gene glnA which is sourced from saccharomyces cerevisiae and has a tyrosine residue mutation at the 99th site on the surface and an sRNA-MicC-EcHfq regulation module of a targeted adenylyltransferase gene glnE into a vector plasmid; the method comprises the following steps: constructing a recombinant plasmid for removing adenosine acylation of glutamine synthase; and transferring the recombinant plasmid into corynebacterium glutamicum of which the odhA gene expression is controlled by a growth stage specific promoter Pcg2705, so as to obtain a corynebacterium glutamicum engineering strain for producing L-glutamine. According to the invention, through multi-gene synergistic modification and space-time metabolic flux optimization, the problem of'growth-synthesis' tradeoff in the prior art is solved, the fermentation efficiency is improved, the fermentation process is simple, and the application prospect is good.

The invention discloses a screening and expression platform construction method for non-essential gene loci of corynebacterium glutamicum, and belongs to the technical field of biology. The center sites CGLRS02940, CGLRS03085, CGLRS08555, CGLRS09020 and CGLRS13560 of the corynebacterium glutamicum identified by the invention have the remarkable advantages that when gene knockout or knockin operation is carried out, the sites do not interfere with the normal physiological metabolism process of strains, and stable expression of exogenous genes can be guaranteed; meanwhile, the gene editing efficiency is relatively high, and an editing target with high universality and high reliability is provided for genetic engineering modification of corynebacterium glutamicum.

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 application discloses a construction method and application of a genetically engineered corynebacterium glutamicum for synthesizing a bacteriochlorophyllide amino acid shinorine, and belongs to the field of genetic engineering and fermentation technology. The genetically engineered corynebacterium glutamicum is constructed by taking corynebacterium glutamicum ATCC 13032 as a host strain, knocking out a competitive pathway gene, integrating a key enzyme gene in multiple copies on a genome, knocking out an acid production pathway gene and a serine dehydrogenase gene, and expressing a gene cluster for synthesizing shinorine on a plasmid, so that the synthesis yield of shinorine in a microbial cell is improved. The strain can produce shinorine in a flask, and 1.12 g / L of shinorine can be accumulated in 72 hours. In fed-batch fermentation in a 50 L fermenter, the strain can be cultured to 8.6 g / L.

The invention relates to Corynebacterium glutamicum with high indigo yield as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The recombinant corynebacterium glutamicum can be used for overexpression of the following genes: a D-glucose-6-phosphate dehydrogenase gene zwf, a II type deoxyarabinose type heptulose phosphate synthase gene aroG and a flavin-containing monooxygenase gene CcFMO. The recombinant corynebacterium glutamicum can weaken the following genes: a pyruvate kinase gene pyk and a tryptophan synthase gene trpB. The corynebacterium glutamicum is used for overexpressing a zwf gene, an aroG gene and a CcFMO gene, weakening a pyk gene and a trpB gene and enhancing the expression quantity of an indole synthetic pathway related enzyme gene, so that the yield of the corynebacterium glutamicum at a shake flask stage reaches 619mg / L in a fermentation production process of indigo under the condition of not additionally adding tryptophan. The invention provides the recombinant corynebacterium glutamicum with high indigo yield, which is low in production cost, good in stability and high in synthesis efficiency, and is suitable for practical popularization and application.

The invention discloses genetically engineered corynebacterium glutamicum and an application of the genetically engineered corynebacterium glutamicum in preparation of ectoine, and the genetically engineered corynebacterium glutamicum contains an L-diaminobutyric acid transaminase gene ectB, an L-diaminobutyric acid acetyltransferase gene ectA and an ectoine synthetase gene ectC, the invention relates to an aspartic acid transferase gene aspB, an aspartic acid kinase gene lysC and an aspartic acid semialdehyde dehydrogenase gene asd, wherein the nucleotide sequences of the gene aspB, the gene lysC and the gene asd are respectively as shown in SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6. The invention further discloses a preparation method of the aspartic acid transferase gene aspB, the aspartic acid kinase gene lysC and the aspartic acid semialdehyde dehydrogenase gene asd. After the recombinant corynebacterium glutamicum constructed by the invention is fermented for 60 hours, the yield of ectoine can reach 50g / L. The genetically engineered corynebacterium glutamicum disclosed by the invention has important practical significance on industrial production and large-scale application of ectoine.

This invention discloses a method for the efficient synthesis of gastrodin using recombinant Corynebacterium glutamicum, belonging to the fields of genetic engineering, fermentation engineering, and synthetic biology. This invention uses Corynebacterium glutamicum as the host bacterium and knocks out… pobA The gene was expressed, and the engineered strain WN01 / pB6 was constructed using the expression plasmid pEC-XK99E to express cladoid-pyruvate lyase, carboxylic acid reductase, and glycosyltransferase, achieving heterologous synthesis of gastrodin. Further overexpression was then performed. aroG Genes, overexpression of protein-coding genes that activate carboxylic acid reductase sfp The engineered strain WN05 / pB6 was able to synthesize 7.37 g / L of gastrodin at the shake flask level, and the gastrodin yield reached 17.77 g / L at 72 h of batch fermentation in a 5-L fermenter, with a yield of 0.247 g / L / h. This provides a brand-new biomanufacturing platform for the industrial production of gastrodin.

The present invention provides recombinant Corynebacterium glutamicum genetically engineered to introduce the N-acetylmannosamine (ManNAc) synthesis pathway, and genetically engineered to express an exogenous N-acetylneuraminic acid synthetase, an exogenous CMP-N-acetylneuraminic acid synthetase, an exogenous lactose transporter, and an exogenous sialyltransferase. The invention also provides a method for producing sialic acid lactose by using the recombinant corynebacterium glutamicum.

The application discloses an expression vector of a biosynthetic metabolic pathway gene of 2-azacyclohexanone and a synthesis method of 2-azacyclohexanone. The expression vector comprises a first promoter and, sequentially inserted downstream of the first promoter, davA-davB genes or davB-davA genes, a Pb promoter or a mutant thereof, an act gene or an orf26 gene or a caiC gene, a ChnR gene or a mutant thereof. The expression vector is further transferred into glutamic acid corynebacterium with high lysine yield, and is subjected to fermentation culture to efficiently synthesize 2-azacyclohexanone. The application synthesizes 2-azacyclohexanone in glutamic acid corynebacterium for the first time, and utilizes an optimized 2-azacyclohexanone molecular probe system to start the expression of key genes in the 2-azacyclohexanone biosynthetic metabolic pathway for the first time, so that high yield of 2-azacyclohexanone is realized. The synthesis method reduces the production cost required by using lysine as a synthesis raw material, eliminates a rate-limiting step, and greatly improves the biosynthesis yield of 2-azacyclohexanone.

As one aspect, the present disclosure provides a bacterium of the genus Corynebacterium capable of producing isopropanol with high productivity. As another aspect, the present disclosure provides a method which enables the production of isopropanol with high productivity while suppressing the production of by-products.　In one aspect, the present disclosure relates to a bacterium of the genus Corynebacterium capable of producing isopropanol, the bacterium being produced by introducing into a bacterium of the genus Corynebacterium a specific gene (A) that encodes an enzyme having an activity of producing acetoacetic acid from acetoacetyl-CoA and / or a specific gene (B) that encodes an enzyme having an activity of producing acetone from acetoacetic acid in a manner that enables expression thereof. In another aspect, the present disclosure relates to a method for producing isopropanol, the method comprising: allowing a genetically modified bacterium of the genus Corynebacterium capable of producing isopropanol to carry out a fermentation reaction in a fermentation tank to produce a fermentation product containing isopropanol; continuously or intermittently withdrawing a portion of a reaction solution containing the fermentation product from the fermentation tank during the fermentation reaction; separating isopropanol from the withdrawn reaction solution by pervaporation; and supplying to the fermentation tank the reaction solution from which isopropanol has been separated.

The invention discloses a method for reducing the molecular weight of gamma-PGA based on a polycistron expression vector, which comprises the following steps: cloning a gamma-polyglutamic acid synthetase gene cluster pgsBCA of bacillus licheniformis into corynebacterium glutamicum F343 with high yield of glutamic acid in a polycistron expression mode for exogenous expression to obtain a recombinant strain pBCA; compared with a recombinant strain pBpCpA expressed by a monocistron, the recombinant strain pBCA carrying a polycistron expression system has the advantage that the molecular weight of gamma-PGA generated by the recombinant strain pBCA is reduced by 15.98%. Therefore, the polycistron expression system is an effective method for reducing the molecular weight of the gamma-PGA.

A non-coding sRNA in Corynebacterium glutamicum and the use thereof. The non-coding sRNA has an RNA sequence having at least 90% or more sequence homology to a transcription product of the DNA sequence as shown in SEQ ID NO: 1. It is verified by means of experiments that the over-expression of the non-coding sRNA in a recombinant bacterium can significantly improve the yield of branched-chain amino acids, such as L-leucine, L-isoleucine and L-histidine, and the sugar-acid conversion rate, and also reduce the generation of by-product acid impurities. In terms of materials, relatively inexpensive molasses is selected, which provides rich carbon and nitrogen sources, to replace corn syrup which has more impurities. Moreover, the molasses is converted by means of enzymolysis into a nutrient substance containing more carbon and nitrogen sources that can be utilized by the strain. In addition, in order to control the carbon-to-nitrogen source ratio in a fermentation broth, a more accurate feeding process is selected, which can not only ensure the fermentation requirements of the strain, but can also effectively prevent excessively high or low nitrogen source levels in the fermentation broth, thereby achieving the aims of increasing the yield and the sugar-acid conversion rate, and reducing the by-product acid impurities.

The present application relates to a kind of acetolactate synthase and its application, belong to enzyme engineering and metabolic engineering technical field.The present application is based on the wild-type acetolactate synthase from Corynebacterium glutamicum (Corynebacterium glutamicum) Corynebacterium glutamicum ) ATCC13032, the encoding gene ilvBN Of it is carried out error-prone PCR, screening obtains acetolactate synthase mutant IlvBN M1 And IlvBN M2 , above-mentioned mutant removes L-valine, L-leucine and L-isoleucine on its feedback inhibition, under the condition that L-valine, L-leucine and L-isoleucine concentration is 0-30 mmol / L respectively, enzyme activity has no obvious change, can be widely used in the synthesis of L-valine, L-leucine and L-isoleucine.

Provided are a novel gluconate repressor variant, a polynucleotide comprising the variant of the present disclosure, a microorganism of the genus Corynebacterium comprising the variant or polynucleotide of the present disclosure, and a method of producing L-arginine using the microorganism of the present disclosure.

A personal cleansing composition for use in a method of inhibiting or killing the growth of bacteria on human skin wherein the bacteria are selected from the group consisting of: Escherichia coli having a pH of up to or equal to 4 in solution, dermatobacterium acnes, and Corynebacterium minutus; wherein the personal cleansing composition comprises a salt of an alkenyl sulfate having an alkenyl chain, or an isomer thereof, or a mixture thereof, wherein the alkenyl chain comprises 18 to 20 carbon atoms.

The application discloses a CgGlmS mutant and application thereof in synthesis of N-acetylglucosamine (GlcNAc) in corynebacterium glutamicum, and belongs to the technical field of biotechnology. In-depth rational analysis and semi-rational modification are carried out on the bottleneck enzyme CgGlmS in the N-acetylglucosamine synthesis pathway. Through systematic research on the structure and function of the original CgGlmS enzyme from corynebacterium glutamicum S9114, a high-efficiency CgGlmS mutant A253M is successfully screened, and the enzyme activity, stability and catalytic efficiency of the mutant are obviously improved compared with those of the wild type. The mutant CgGlmS is overexpressed in corynebacterium glutamicum S9114 by using a plasmid, and the yield of N-acetylglucosamine in corynebacterium glutamicum is improved. Through the directional modification of the CgGlmS enzyme, the catalytic efficiency is improved, and the accumulation of N-acetylglucosamine in corynebacterium glutamicum is promoted, which provides strong support for the efficient production of N-acetylglucosamine.

The present invention relates to a culture medium composition containing sugars and amino acids derived from human skin, microorganisms of the genera Staphylococcus, Streptococcus, Cutibacterium, Corynebacterium, and Rothia, and a cosmetic composition containing their lysate or culture medium. The mixed culture medium of human skin-derived microorganisms contained in the cosmetic composition is cultured in the culture medium composition and exhibits a significantly superior skin-improving effect compared to each of the individual culture mediums.

The application discloses a screening method and application of a Corynebacterium glutamicum neutral site, and relates to the technical field of biology.The application also provides an application of the neutral site in expression of a target protein or polypeptide in the Corynebacterium glutamicum, wherein the application is that a coding gene of the target protein or polypeptide is integrated on the neutral site; and the nucleotide sequence of the neutral site is shown in any one of SEQ ID NO.1 to SEQ ID NO.5.The neutral site of the Corynebacterium glutamicum identified in the application has the following important characteristics: when a gene editing operation (such as gene knockout or knock-in) is performed, the site can ensure that normal physiological metabolism of the strain is not affected, and can maintain efficient and stable expression of an exogenous gene; meanwhile, the site shows excellent editing efficiency, and provides an ideal target site with good universality and strong reliability for genetic modification of the Corynebacterium glutamicum.

The application discloses a method for preparing functional caragana fiber compound feed and belongs to the technical field of feed processing. The method comprises the following steps: taking caragana as raw material, performing solid-liquid separation after dilute sulfuric acid pretreatment, preparing cordycepin and lysine through liquid state fermentation of cordyceps militaris and glutamic acid corynebacterium respectively, neutralizing caragana solid fiber and caragana fiber raw material obtained through ammonia treatment, and preparing fiber feed through solid anaerobic fermentation of the compound fermentation strain, and finally, compounding the three to obtain functional caragana fiber compound feed. The method realizes efficient utilization of all components of caragana sugar and fiber resources, and the prepared compound feed is rich in cordycepin, lysine and dietary fiber, the crude protein content of the fiber feed is more than 20%, and the fiber feed is rich in cordycepin and lysine functional components. The method not only provides a new way for resource utilization and high value utilization of caragana, but also provides a new idea for research and development of functional coarse feed, and has good economic, ecological and industrial popularization value.

A novel technique for improving secretory production of a heterologous protein by coryneform bacteria is provided, and thereby a method for secretory production of a heterologous protein is provided. A coryneform bacterium having an ability of secretory producing a heterologous protein and having been modified so that an activity of a Mdh protein is reduced is cultured to produce the heterologous protein by secretory production.

The invention relates to the technical field of construction of recombinant bacteria, in particular to recombinant corynebacterium glutamicum for producing L-ornithine as well as a construction method and application of the recombinant corynebacterium glutamicum for producing L-ornithine, corynebacterium glutamicum SO30 is taken as an original strain, a SigD gene and a FruR gene are sequentially knocked out through a suicide plasmid pK18mobsacB mediated homologous recombination technology, and double knockout recombinant corynebacterium glutamicum SO30-SigD-FruR is constructed. The L-ornithine yield of the strain subjected to shake-flask fermentation reaches 51.88 g / L and is increased by 56.7% compared with that of an original strain SO30; the fermentation yield of a 5L bioreactor reaches 123 g / L and is increased by 57.6% compared with that of an original strain SO30, and the cell biomass and the sugar conversion rate are remarkably increased. Through global transcriptional regulation factor transformation, the existing yield bottleneck is broken through, and efficient strains and technical support are provided for industrial fermentation production of L-ornithine.

This invention belongs to the field of biotechnology. To improve the effectiveness of biological control of plant diseases, this invention discloses *Bacillus velezensis* ZF516 and its application in controlling various soil-borne diseases. The *Bacillus velezensis* strain number is ZF516, and its registration number at the China General Microbiological Culture Collection Center is CGMCC No. 30136. This *Bacillus velezensis* strain can prevent and / or treat tomato bacterial canker, tomato bacterial wilt, or cowpea root rot, or can be used to prepare products for the prevention and / or treatment of these diseases. This strain also exhibits good inhibitory effects against *Fusarium oxysporum*, *Diplostomum spp.*, *Rhizoctonia solani*, *Fusarium solanum*, *Corynebacterium multiflorum*, *Botrytis cinerea*, *Agrobacterium tumefaciens*, *Pectinobacter brasiliensis*, *Ralstonia solanacearum*, *Streptomyces spp.*, etc.

The present invention relates to a Corynebacterium mutant microorganism that produces L-glutamic acid and a method for producing L-glutamic acid using the same, and more specifically to a novel mutant, polynucleotide, and transformant of pantoate-beta-alanine ligase involved in the L-glutamic acid biosynthetic pathway, and a method for producing L-glutamic acid using the same. The pantoate-beta-alanine ligase mutant of the present invention has altered enzymatic activity due to the substitution of one or more amino acids in the amino acid sequence constituting pantoate-beta-alanine ligase, and a recombinant microorganism containing the same can efficiently produce L-glutamic acid.

The invention belongs to the technical field of plant biology, and particularly provides application of a CsTSR gene in cucumber corynespora leaf spot resistance. A transient expression strain and a cstsr mutant are respectively obtained through transient transformation of the CsTSR gene and a Tnt1 reverse transcription transposon mutant library, and the CsTSR gene is found to positively regulate the resistance of the cucumber to the corynespora leaf spot disease. The miR164d can regulate and control the CsTSR transcription factor in a targeted manner. A dual luciferase report and a GUS (glucuronidase) staining experiment result show that the miR164d negatively regulates and controls the CsTSR, and the CsTSR-mediated cucumber disease resistance can be enhanced by silencing the miR164d. The invention discloses a functional mechanism of the miR164d-CsTSR molecular module in cucumber pathogenic bacterium infection resistance, and provides a new reference gene resource for cultivation of cucumber disease-resistant varieties.

The invention relates to the technical field of biochemical engineering, in particular to recombinant corynebacterium glutamicum for producing 5-aminolevulinic acid as well as a construction method and application of the recombinant corynebacterium glutamicum. According to the construction method of the recombinant corynebacterium glutamicum capable of producing 5-aminolevulinic acid, a hemARsp gene is heterologously expressed, genes such as SigB and RamB are knocked out and optimized, transcriptional regulatory factors related to carbon metabolism, nitrogen metabolism, phosphorus metabolism, SOS response / pressure response, iron and sulfur homeostasis and the like in the recombinant corynebacterium glutamicum are optimized, and the recombinant corynebacterium glutamicum capable of producing 5-aminolevulinic acid is obtained. And carrying out global regulation and control on a cell metabolism network, and guiding the cell metabolism flux to the accumulation direction of a target product, so as to obtain the recombinant corynebacterium glutamicum for high yield of 5-aminolevulinic acid.

The invention discloses a construction method of corynebacterium glutamicum for efficient de novo synthesis of 1, 5-pentanediamine, and belongs to the technical field of biochemical engineering. The recombinant corynebacterium glutamicum is obtained by taking corynebacterium glutamicum CGlys for producing lysine as a starting strain, optimizing codons and controlling expression by using an optimal promoter Psod; when the recombinant strain is used for fermentation synthesis, 1, 5-pentamethylene diamine can be produced by using glucose under mild conditions, and the recombinant strain has a very wide application prospect.

The present invention relates to a Corynebacterium mutant strain that has improved glycine-producing ability compared to a parent strain through mutagenesis using irradiation, a method for producing the strain, and a method for producing glycine using the strain.The Corynebacterium mutant strain has improved glycine-producing ability compared to a parent strain, and glycine can be produced with high efficiency and high yield using the strain.

The invention provides an attenuated pertussis CS recombinant bacterium, the CS recombinant bacterium is obtained by mutating a ninth arginine residue of an S1 subunit in a CS strain with a preservation number of CMCC 58003 into a lysine residue and mutating a 129th glutamic acid residue into a glycine residue, and the amino acid sequence of the S1 subunit of the CS recombinant bacterium is as shown in SEQ ID NO.1. The invention further provides a preparation method of the attenuated pertussis CS recombinant bacterium. The method for constructing the recombinant bacteria comprises the following steps: constructing recombinant plasmids; preparing competent cells of the wild pertussis CS strain; electrically transferring the recombinant plasmid into competent cells of the wild pertussis CS strain; respectively preparing BG sheep blood screening plates containing Amp and DOG; sequentially screening a first round and a second round of recombinant pertussis CS strains; and preparing the two-round recombinant pertussis CS strain into a competent strain, and screening twice according to the same steps to obtain the attenuated pertussis CS recombinant strain. The constructed recombinant bacteria can efficiently obtain detoxified PT, and can be applied to products for treating, detecting and preventing pertussis diseases.

The invention discloses a method for promoting recombinant corynebacterium glutamicum to secrete chondroitin oligosaccharide by changing cell membrane permeability, and belongs to the field of fermentation engineering. According to the invention, a high-copy double-plasmid system is used in corynebacterium glutamicum for coupling synthesis and cutting of chondroitin through a genetic engineering means, so that the yield of chondroitin oligosaccharide synthesized by corynebacterium glutamicum is increased. In order to promote the secretion of chondroitin oligosaccharide, the permeability of cell membranes is increased by quantitatively and regularly adding a surfactant tween-40 in the fermentation process. Through high-copy plasmid expression, culture medium optimization, surfactant treatment and other strategies, the yield of chondroitin oligosaccharide generated by fermentation of the constructed recombinant corynebacterium glutamicum is increased by 37.5%.

Provided is a recombinant Corynebacterium glutamicum, which is genetically engineered to introduce an N-acetyl-mannosamine (ManNAc) synthetic pathway, and is genetically engineered to express a heterologous N-acetylneuraminic acid synthase, a heterologous CMP-N-acetylneuraminic acid synthase, a heterologous lactose transporter, and a heterologous sialyltransferase. Provided is a method for using the recombinant Corynebacterium glutamicum to produce sialyllactose.