168 results about "Corynebacterium" patented technology

The invention belongs to the technical field of biological medicine, and particularly relates to application of corynebacterium casei in preparation of medicine for preventing and treating lung injury. By constructing a radon inhalation-induced lung injury mouse model, after exposure at different radon levels, the weight of a mouse is reduced, the lung coefficient is increased, the tidal volume of the lung, the ventilation volume per minute and the maximum inspiration flow are reduced, the respiratory function is inhibited, the alveolar cavity of the mouse is abnormally expanded, the pulmonary septum is thickened and fractured, and more fibrin exudation is accompanied; the traditional Chinese medicine composition can be used for effectively inhibiting pulmonary fibrosis and lung injury due to obvious inflammatory infiltration beside bronchial tubes, extravasated blood in small blood vessels and lung tissue collagen deposition, and can be used for gavage of corynebacterium casei to effectively inhibit the above phenomena. Therefore, the corynebacterium casei can be used for preventing and treating lung injury, especially lung injury caused by radon exposure.

The embodiment of the invention discloses a recombinant III-type humanized collagen as well as a preparation method and application thereof. The amino acid sequence of the recombinant III-type humanized collagen is shown as SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3. The recombinant III-type humanized collagen has the advantages that the recombinant III-type humanized collagen can be used for preparing the recombinant III-type humanized collagen; the recombinant III-type humanized collagen provided by the invention has the activity of promoting cell adhesion, the amino acid sequence of the recombinant III-type humanized collagen is selected from the amino acid sequence of natural collagen, and the recombinant III-type humanized collagen does not generate immune response when being applied to a human body. The characteristic that corynebacterium glutamicum does not secrete endotoxin is utilized, the preparation method is simple, the product can meet the requirements of food, cosmetics, medical instruments and drugs without complex separation, and reference is provided for low-cost industrial production.

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 corynebacterium glutamicum whole genome random mutation tool and application thereof, and belongs to the technical field of genetic engineering and biological mutagenesis. DsDNA is unwound in the transcription process based on DNA helicase, DNA adenine deaminase and the DNA helicase are fused, and whole-genome random mutation of corynebacterium glutamicum is achieved. The whole genome random mutation method disclosed by the invention is applied to mutagenesis of an original strain of Corynebacterium glutamicum ATCC 13032, and a mutant strain of which the biomass is 1.95 times of that of WT (OD600 is 2.2) and 1.99 times of that of WT / pXMJ19 (OD600 is 2.16) when being cultured under the condition that the pH value is 5.5 is obtained. The corynebacterium glutamicum whole genome random mutation method disclosed by the invention has important guiding significance for screening other mutation strains with enhanced robustness.

The invention discloses a CgGlmS mutant and application of the CgGlmS mutant in synthesis of N-acetylglucosamine (GlcNAc) from corynebacterium glutamicum, and belongs to the technical field of biology. According to the invention, a bottleneck enzyme CgGlmS in a synthetic route of N-acetylglucosamine is subjected to deep rational analysis and semi-rational transformation. Through systematic research on the structure and function of an original CgGlmS enzyme from corynebacterium glutamicum S9114, the efficient CgGlmS mutant A253M is successfully screened out, and the enzyme activity, stability and catalytic efficiency of the mutant are obviously improved compared with those of a wild type. The mutant CgGlmS is overexpressed in corynebacterium glutamicum S9114 by using plasmids, so that the yield of N-acetylglucosamine in corynebacterium glutamicum is increased. The CgGlmS enzyme is directionally modified, so that the catalytic efficiency of the CgGlmS enzyme is improved, accumulation of N-acetylglucosamine in corynebacterium glutamicum is further promoted, and powerful support is provided for efficient production of the corynebacterium glutamicum.

The invention discloses a construction method of recombinant corynebacterium glutamicum for fermentation synthesis of poly beta-(1, 6)-N-acetylglucosamine polysaccharide, and belongs to the technical field of bioengineering. The recombinant corynebacterium glutamicum capable of preparing the PNAG through fermentation is constructed by using synthetic biology and genetic engineering technologies for the first time, cheap carbon sources and nitrogen sources can be utilized, the recombinant corynebacterium glutamicum is suitable for high-density fed-batch fermentation, and the generation cost of the PNAG is reduced; the yield after 42 hours of fermentation can reach 6.82 g / L, and the method has industrial application potential.

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.

The present invention belongs to the field of biomedicine technology, and in particular to the application of Corynebacterium casei in the preparation of medicines for preventing and treating lung injury. The present invention constructs a mouse model of lung injury caused by radon inhalation and finds that after exposure to different radon levels, the mice lose weight, increase the lung coefficient, reduce the lung tidal volume, minute ventilation and maximum inspiratory flow, and their respiratory function is inhibited. The alveolar cavity of the mice is abnormally expanded, the lung septum is thickened and fractured, accompanied by more fibrin exudation, obvious inflammatory infiltration appears near the bronchus, congestion occurs in small blood vessels, and collagen deposition in lung tissue can lead to the occurrence of pulmonary fibrosis, causing lung injury. Oral administration of Corynebacterium casei can effectively inhibit the above-mentioned phenomenon. Therefore, Corynebacterium casei can be used for the prevention and treatment of lung injury, especially lung injury caused by radon exposure.

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 belongs to the technical field of biochemical engineering, and provides a method for improving the high-temperature tolerance of corynebacterium glutamicum and the production capacity of L-tryptophan. The invention provides a novel high-temperature tolerance related gene target osrR of corynebacterium glutamicum, biomass accumulation of corynebacterium glutamicum under a high-temperature condition is remarkably improved by regulating and controlling the expression level of the novel high-temperature tolerance related gene target osrR, and the fermentation yield of L-tryptophan of recombinant corynebacterium glutamicum under the high-temperature condition is improved. Specifically, by reducing the expression level of the recombinant corynebacterium glutamicum, the biomass accumulation of the recombinant corynebacterium glutamicum under the condition of 40 DEG C and the yield of the L-tryptophan under the condition of 33.5 DEG C are remarkably improved, and support is provided for high-temperature fermentation production of the L-tryptophan. Wide application prospects are realized.

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 relates to the technical field of biology, and provides a bacterial strain capable of accelerating and promoting germination of tulipa seeds and application of the bacterial strain, the bacterial strain is obtained by being separated from soil of tulipa germinating seedlings, the bacterial strain is identified as aspergillus niger through ITS sequence sequencing after being purified, the taxonomic name is Aspergillus niger, and the preservation number of the bacterial strain is CCTCC NO: M 2025709. Through an associated germination test of the strain and tulipa plant seeds, the result shows that the strain can obviously accelerate the germination of the tulipa seeds and improve the germination rate of the tulipa seeds.

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 present invention relates to a novel promoter derived from Corynebacterium and to a use of a variant thereof in the production of a protein or a rare sugar. In the present invention, a sod gene promoter derived from Corynebacterium ammoniagenes, which enables regulation of gene expression and exhibits significantly stronger activity compared to conventionally used promoters, was developed, promoter variants having various strengths were identified from the developed promoter, and by using the identified improved promoters, two enzymes capable of converting a common sugar into a rare sugar were highly expressed in a single vector system, thereby enabling production of a rare sugar via a one-pot reaction. Accordingly, the promoters can be utilized for the production of a target protein or a rare sugar.

The invention relates to the technical field of tag peptides, and particularly discloses a tag peptide combined on the surface of corynebacterium, and the amino acid sequence of the tag peptide is shown as SEQ ID NO: 1 or SEQ ID NO: 2. The tag peptide provided by the invention enriches the types of tag peptides bound to the surfaces of current bacteria, and corynebacterium bacteria bound with the tag peptide have wider adaptability to pH and can be compatible with the optimal activity conditions of more foreign proteins; the molecular weight of the tag peptide is only 3.5 kDa, so that the interference to the structure or function of an exogenous protein can be remarkably reduced; in addition, the tag peptide can be fused at the N end or the C end of the target protein, the binding efficiency is not affected, simultaneous display of multiple proteins is supported, and a basis is provided for multi-enzyme cascade reaction and multivalent vaccine development.

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 invention belongs to the technical field of biochemical engineering, and provides a method for increasing the yield of L-tryptophan of corynebacterium glutamicum. The invention provides a new gene target gluA closely associated with synthesis of L-tryptophan in corynebacterium glutamicum, the yield of L-tryptophan of recombinant corynebacterium glutamicum is remarkably increased by knocking out the gene, and meanwhile, the invention also provides a method for increasing the yield of L-tryptophan of recombinant corynebacterium glutamicum by coupling gluA knockout, glnK knockout and sugR overexpression. By regulating and controlling the expression level of the gluA gene in the recombinant corynebacterium glutamicum for high yield of the L-tryptophan and coupling with expression regulation and control of glnK and sugR, the concentration of the L-tryptophan in fermentation liquor is remarkably improved, efficient synthesis of the L-tryptophan in the recombinant corynebacterium glutamicum is achieved through an optimized fermentation scheme, and the L-tryptophan has very wide application prospects.

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 invention discloses a strain for the one-step production of N-methylpyrrolidone, belonging to the technical field of genetic engineering. By knocking out the metabolic pathway of the precursor GABA and some branch pathways, and enhancing the expression of some key genes in the synthesis pathway, the invention constructs an engineered Corynebacterium glutamicum with increased GABA production as the chassis cell for N-methylpyrrolidone synthesis. The invention expresses an N-methyltransferase derived from Coffea canephora in the chassis cell, enabling the constructed strain to ferment N-methylpyrrolidone with glucose as the carbon source in an environment with a pH of 7.0, and the production can reach 5.28 g / L after 96 h of fermentation.

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.