61 results about "Corynebacterium" patented technology

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 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.

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 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 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 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.

The present invention relates to the technical field of microbial metabolic engineering, and in particular to a Corynebacterium glutamicum strain capable of efficiently producing D-pantothenic acid, a construction method therefor, and the use thereof. The construction method specifically comprises the following steps: (1) using wild-type Corynebacterium glutamicum ATCC 13032 as a chassis, attenuating ilvA and performing mutation expression so as to construct DPA1; (2) knocking out pyruvate quinone oxidoreductase and inserting a panBCE gene expression cassette into same so as to construct DPA2; (3) introducing into the DPA2 an acetohydroxy acid synthase, an acetolactate isomeroreductase and a 2-hydroxyacid dehydratase which are overexpressed by using a pEC-xk99E overexpression plasmid and derived from Escherichia coli W3110 and Bacillus subtilis 168, and are endogenous and are obtained by screening; and (4) under the control of a strong promoter, subjecting the selected ilvBNCD gene to tandem overexpression by means of pEC-xk99E to construct DPA3, so that by enhancing the driving force in a principle synthesis pathway of pantothenic acid and upstream pathways, the Corynebacterium glutamicum strain capable of efficiently producing D-pantothenic acid is obtained. The strain provided by the present invention can use carbon sources to produce D-pantothenic acid with a high yield and high conversion rate.

The present disclosure relates to a microorganism of the genus Corynebacterium producing L-amino acid, a method for producing L-amino acid using the same, use of L-amino acid production, and a composition for producing L-amino acid.

The invention relates to airway medicaments. Described herein are compositions, methods, kits, and devices for treating and / or preventing a wide range of disease conditions. In particular, the bacterial populations described herein are live, purified bacteria for use in modulating, restoring, and / or promoting microbiome in the upper respiratory tract, including the nasal cavity, of a subject to promote health. Such bacterial populations may comprise a single or multiple bacterial strains. The plurality of bacterial strains may be strains from the same or different species, including Corynebacterium and / or Cunglada lazy.

The present invention relates to a Corynebacterium glutamicum mutant strain having increased L-glutamic acid productivity, a method for constructing the same, and a method of producing L-glutamic acid using the same. The Corynebacterium glutamicum mutant strain is a strain into which a mechanosensitive ion channel gene derived from a Corynebacterium sp. strain has been introduced, and thus it can produce L-glutamic acid in an improved yield due to enhancement of glutamic acid release. Therefore, when the mutant strain is used, it is possible to more effectively produce L-glutamic acid.

The application discloses a recombinant type III humanized collagen, a preparation method and application thereof, and an amino acid sequence of the recombinant type III humanized collagen is shown in SEQ ID NO. 1, SEQ ID NO. 2 or SEQ ID NO. 3. The recombinant type III humanized collagen provided in the application has cell adhesion promoting activity, the amino acid sequence of the recombinant type III humanized collagen is selected from natural collagen amino acid sequences, and the application of the recombinant type III humanized collagen to human body does not cause an immune response. The application utilizes the characteristics that corynebacterium glutamicum does not secrete endotoxins, the preparation method is simple, and the product can meet the requirements of cosmetics, medical devices and medicines without complicated separation, and thus provides a reference for low-cost industrial production.

This application discloses the application of the pyc gene and its mutants in the preparation of lysine, belonging to the field of genetic engineering technology. The technical problem this application aims to solve is: how to increase the amino acid yield of microorganisms. To solve the above technical problem, this application provides a protein obtained by mutating serine (S) at position 1059 of pyruvate carboxylase to proline (P) and related biological materials. This application also provides the application of said protein and related biological materials in constructing recombinant bacteria and preparing L-amino acids. This application has obtained a mutant pyruvate carboxylase (pyc) that increases the lysine yield in target microorganisms through extensive experiments. S1059P The results of the recombinant bacterial validation showed that, compared with the original target microorganism and recombinant microorganisms that overexpress wild-type pyruvate carboxylase (e.g., recombinant Corynebacterium glutamicum), the L-lysine production of recombinant Corynebacterium glutamicum expressing this mutant pyruvate carboxylase was significantly increased.

The present disclosure relates to a Corynebacterium sp. mutant strain, a method for producing the strain, and a method for producing glycine using the strain.

This invention discloses the promoter EP11 and related biomaterials and applications, belonging to the field of biotechnology. The technical problem solved by this invention is how to increase the content of L-lysine. The promoter EP11 disclosed in this invention has the sequence shown in SEQ ID No. 1 of the sequence listing. The EP11 promoter is introduced into *Corynebacterium glutamicum*, a bacterium capable of synthesizing L-lysine, thereby driving the expression of genes in the L-lysine synthesis pathway, resulting in recombinant *Corynebacterium glutamicum*. Culturing this recombinant *Corynebacterium glutamicum* yields L-lysine, and the production of L-lysine is increased. This demonstrates that the EP11 promoter of this invention not only possesses promoter activity but can also be used for the production of L-lysine.

The invention discloses application of a succinate dehydrogenase mutant in glutamic acid production, and belongs to the technical field of genetic engineering. In order to improve the glutamic acid yield of microorganisms, the invention provides a succinate dehydrogenase mutant, and the succinate dehydrogenase mutant at least comprises protein obtained by mutation of the 301st amino acid residue site of succinate dehydrogenase. Recombinant bacterium verification results show that compared with original target microorganisms, the glutamic acid yield of recombinant corynebacterium glutamicum expressing the succinate dehydrogenase mutant or overexpressing the succinate dehydrogenase is remarkably increased, and an unexpected technical effect is achieved.

This invention discloses a strain of Corynebacterium glycinophilus T68 that produces nematicidal active compounds and its applications. In a pot experiment for controlling root-knot nematodes, its control efficacy reached 62.03%. The LC50 activity of the compound phenylacetamide produced by this strain against southern root-knot nematodes was [not specified in the original text]. 50 The concentration was 122.58 μg / ml, demonstrating good control efficacy against root-knot nematodes. This actinomycete can be effectively used to control root-knot nematode disease. It has great application potential in solving root-knot nematode damage in vegetable production.

Provided is a composition for preventing or ameliorating bromhidrosis, including a seed fermentation product of a plant belonging to the genus Lotus. The composition may inhibit production of substances related to bromhidrosis by reducing proportions of microorganisms of the genus Corynebacterium and the genus Anaerococcus, which are related to bromhidrosis, in the microbiome.

This application discloses the application of the aspB gene and its mutants in the preparation of lysine, belonging to the field of genetic engineering technology. The technical problem this application aims to solve is: how to increase lysine production in genetic engineering. To solve the above technical problem, this application provides a protein containing the amino acid sequence described in SEQ ID NO: 4 and its application in increasing microbial lysine production. This application has obtained a mutant aspartate transaminase (aspB) with high lysine expression function through extensive experiments. D332N The results of the recombinant bacterial validation showed that, compared with the original target bacteria and the recombinant Corynebacterium glutamicum overexpressing wild-type aspartate transaminase, the L-lysine production of the recombinant Corynebacterium glutamicum expressing this mutant aspartate transaminase was significantly increased.