326 results about "Metabolic engineering" patented technology

The invention relates to a construction method of escherichia coli engineering bacteria with high yield of N-acetylglucosamine. According to the method, the metabolic network of escherichia coli is directionally modified through metabolic engineering, the problems of'product re-decomposition ', by-product accumulation, poor genetic stability and the like of natural strains are solved, and efficient industrial production of GlcNAc is realized.

The invention relates to an L-valine production strain as well as a construction method and application thereof, the strain is obtained by transformation through a metabolic synthesis path of L-valine, and the construction method comprises the following steps: by taking Cornebacterium glutamicum ATCC 13032 (C.glutamicum) as an original strain, carrying out editing transformation such as gene knockout, gene overexpression, gene heterologous expression, gene double-copy expression and the like on a genome of the original strain by utilizing a metabolic engineering means. The L-valine production strain disclosed by the invention can be used for synthesizing L-valine from the beginning by taking glucose as a carbon source, has the advantages of high production rate, short fermentation period, high strain stability and low cost, and lays a foundation for realizing large-scale production of L-valine. According to the fermentation tank fermentation method, the L-valine is produced in a mechanical stirring type fermentation tank by using the L-valine production strain Va1-09, 86.4 g / L of L-valine is produced by fermenting for 36 hours, and the method has a good industrial prospect.

The invention provides a genetically engineered bacterium for producing O-succinyl-L-homoserine as well as a construction method and application of the genetically engineered bacterium. In a chassis bacterium genome, the expression of a 2-ketoglutaric acid decarboxylase encoding gene sucA is enhanced, and the expression of a succinyl-coenzyme A synthetase encoding gene sucD is weakened, so that the supply of succinyl-coenzyme A is increased; the method comprises the following steps: increasing the NADPH (Nicotinamide Adenine Dinucleotide Phosphate) reducing capacity and ATP (Adenosine Triphosphate) energy supply of a chassis bacterium, increasing DNA (Deoxyribose Nucleic Acid) in combination with a transcription dual regulatory factor ompR to improve the stress resistance of escherichia coli under high osmotic pressure, and introducing an overexpression plasmid containing a homoserine transsuccinylase coding gene metA to construct the genetically engineered bacterium for producing O-succinyl-L-homoserine. The engineering strain obtained through a systematic metabolic engineering modification strategy can realize effective accumulation of OSH, the shake flask yield of OSH reaches 19.8 g / L, the fed-batch fermentation yield of a 5L fermentation tank reaches 110.5 g / L, the sugar-acid conversion rate reaches 52.6%, and a foundation is laid for subsequent construction of high-yield OSH engineering bacteria.

The invention discloses a spermidine synthetase mutant and application of the spermidine synthetase mutant in synthesis of spermidine, and belongs to the technical field of genetic engineering and metabolic engineering. According to the method, the spermidine synthetase is mutated firstly, the efficiency of the spermidine synthetase is improved, and on the basis, the dissolved oxygen and fed-batch concentration are optimally controlled through fermentation, so that the yield of spermidine is improved. The corynebacterium glutamicum provided by the invention is an engineering bacterium with plasmids and without defects, and after the corynebacterium glutamicum is subjected to fed-batch fermentation in a 5L fermentation tank for 68 hours, the yield of spermidine reaches 25.1 g / L, and the sugar-acid conversion rate is 17.9%.

The invention relates to the technical field of synthetic biology, metabolic engineering and cell culture. The present invention provides a cell for the production of a negatively charged, preferably sialylated, oligosaccharide wherein the cell is genetically engineered to have or express, preferably overexpress, a hydrolytic UDP-N-acetyl-D-glucosamine-2-epimerase. The invention further provides the use of said cells in culture or incubation. Also described are methods of producing negatively charged, preferably sialylated oligosaccharides using the cells, and purification of the negatively charged, preferably sialylated oligosaccharides.

The invention discloses a yarrowia lipolytica engineering strain with high yield of resveratrol as well as construction and application of the yarrowia lipolytica engineering strain, and belongs to the technical field of synthetic biology and metabolic engineering. The invention provides a yarrowia lipolytica engineering strain. A 4-coumaric acid-CoA ligase gene, a multi-copy resveratrol synthase gene, a transketolase 1 gene, a fusion gene of a histidine phosphate aminotransferase gene and a tyrosine ammonia lyase gene, a glycolytic pathway hexokinase gene, a 6-phosphofructokinase-1 gene and a phosphoglycerate kinase gene are integrated, so that the recombinant protein is obtained. According to the present invention, by using glucose as the substrate, the high yield of resveratrol can be achieved by using glucose as the substrate, the resveratrol yield during shake-flask culture fermentation can achieve 3.157 g / L, the resveratrol yield can be increased to 30.7 g / L during 5L-scale fed-batch fermentation, and the resveratrol yield can be increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L; the maximum value is reported by the yarrowia lipolytica system at present.

The invention belongs to the technical field of metabolic engineering and biological manufacturing, and particularly relates to an engineering bacterium for producing gallic acid (GA) and application of the engineering bacterium in production of gallic acid. According to the method, key metabolic nodes in the gallic acid production process are systematically regulated, for example, the hydroxylase activity is improved, the pyruvic acid flow direction is regulated, the aromatic amino acid precursor supply is improved, and the dynamic balance between the engineering bacterium growth and the gallic acid product synthesis is realized, so that the gallic acid yield and the carbon yield are remarkably improved. Experiments show that the gallic acid yield of the engineering bacteria prepared by the invention reaches 12.5 g / L in a shake flask experiment, and reaches 109.8 g / L in a 3 L fermentation tank; and the carbon yield is increased to 0.64 g / g glucose, which is the highest level reported in the current literature.

The invention relates to recombinant escherichia coli for de novo synthesis of piceatannol as well as a construction method and application thereof, and belongs to the field of synthetic biology and metabolic engineering. According to the invention, an L-tyrosine production strain is used as an original strain, five key genes required for synthesizing piceatannol are combined and connected to a plasmid vector, and a recombinant plasmid is introduced into the original strain, so that an engineering strain for heterologous synthesis of piceatannol is successfully constructed. According to the invention, the yield of piceatannol is further obviously improved through copy number optimization.

Escherichia coli is modified by means of metabolic engineering, a genetically engineered bacterium for high yield of O-succinyl-L-homoserine is obtained, and a construction method of the genetically engineered bacterium comprises the following steps: weakening a synthetic route of an organic acid metabolic byproduct of a chassis strain E.coli W3110 [delta] metI [delta] metJ [delta] thrB [delta] metB; wherein the synthetic route of the organic acid metabolic byproducts is selected from at least one of a formic acid synthetic route, a lactic acid synthetic route and an ethanol synthetic route. The constructed genetically engineered bacterium is applied to microbial fermentation production of the O-succinyl-L-homoserine, effective accumulation of the O-succinyl-L-homoserine is achieved, the sugar acid conversion rate reaches 46.8%, and a foundation is laid for subsequent construction of high-yield O-succinyl-L-homoserine engineering bacteria.

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. This disclosure describes a cell metabolically engineered for production of a mixture of at least three different oligosaccharides. Furthermore, this disclosure provides a method for the production of a mixture of at least three different oligosaccharides by a cell as well as the purification of at least one of the oligosaccharides from the cultivation.

The application provides a recombinant Pichia pastoris strain and a construction method and application thereof. The construction method comprises the following steps: taking a Pichia pastoris strain as a host strain, constructing a double synthesis pathway of phenylalanine ammonia lyase and tyrosine ammonia lyase in the host strain to obtain a first engineering strain; relieving the branch acid feedback inhibition of the first engineering strain based on metabolic engineering to obtain a second engineering strain; optimizing the aromatic amino acid synthesis pathway of the second engineering strain to obtain a third engineering strain; knocking out the branch metabolic pathway of the third engineering strain to obtain a fourth engineering strain; increasing the precursor supply of the fourth engineering strain to obtain the recombinant Pichia pastoris strain. The construction method can systematically optimize the synthesis path of p-coumaric acid, and the recombinant Pichia pastoris strain for efficiently producing p-coumaric acid with methanol as a carbon source is constructed. Compared with the host strain, the yield of p-coumaric acid of the recombinant Pichia pastoris strain is significantly improved.

The invention provides a strain for producing L-isoleucine as well as a construction method and application thereof. The strain is obtained by modifying a chassis strain escherichia coli XX12 by utilizing a metabolic engineering modification method, ldhA, adhE and pflB genes are deleted, the transcriptional levels of aspC, pykF, pntAB, ppK, ppc, ilvAfbr, ilvIHfbr and ygaZH genes are up-regulated, the transcriptional level of a leuA gene is down-regulated, a bcd gene derived from B.subtilis 168 and ppnK and cysK genes derived from Cornebacterium glutamicum ATCC 13032 are heterologously expressed, and the strain has the advantages that the strain can be used for producing L-isoleucine; the method has the advantages of no need of adding resistant substances, good L-isoleucine synthesis capability, short fermentation period by using glucose as a carbon source, and good economic benefit and industrial application value.

The invention discloses application of a gene GlZn2Cys661 in regulation and control of synthesis of ganoderic acid. The nucleotide sequence of the gene GlZn2Cys661 is as shown in SEQ ID NO. 1. The gene expression is regulated and controlled by constructing an overexpression or silent vector, so that the synthesis of the ganoderic acid is promoted or inhibited, and an effective method is provided for regulating and controlling the yield of the ganoderic acid. The invention relates to the technical field of bioengineering, and discloses an application of a gene GlZn2Cys661 in regulation and control of ganoderic acid synthesis. The invention provides a key gene target and a core engineering strain for efficiently producing ganoderic acid through a metabolic engineering means, and has important application value.

The invention belongs to the technical field of biology, and particularly discloses a high-density fermentation method and application of recombinant spider silk protein, and the high-density fermentation method comprises the following steps: firstly, transforming an expression vector carrying spider silk protein genes into modified pichia pastoris cells to obtain a strain; the activated strain is subjected to expanding culture, high-density fermentation culture and purification, and recombinant spider silk protein is obtained; wherein the transformation means of the pichia pastoris cell is to integrate Vitreoscilla hemoglobin Vhb gene into the pichia pastoris cell by utilizing a gene recombination technology, and the expression of the Vitreoscilla hemoglobin Vhb gene is improved by carrying out metabolic engineering transformation on the pichia pastoris cell for expression and optimizing the regulation and control of staged fermentation parameters, so that the expression efficiency of the Vitreoscilla hemoglobin Vhb gene is improved. The problems that in existing recombinant protein fermentation, oxygen dependence is high, product degradation is serious, purity is low, and large-scale stability is poor are solved, the yield and purity of recombinant spider silk protein are guaranteed, and large-scale production of the recombinant spider silk protein is facilitated.

The invention relates to the field of microbial metabolism engineering, and discloses a method for increasing the yield of 2 '-fucosyllactose (2'-FL) through the synergistic effect of various alpha-1, 2-fucosyltransferases. According to the method, two or more alpha-1, 2-fucosyltransferases with different sources are co-expressed in microbial cells, and the complementary enzymatic characteristics of the alpha-1, 2-fucosyltransferases are utilized, so that the synthesis efficiency of the 2 '-FL is remarkably improved, and the yield of the 2'-FL is effectively improved. The microbial cell is escherichia coli C43 (DE3) (E. coli C43 lacZ waaF with lacZ and waaF genes knocked out, the serial number is SL), a pETDuet-1 plasmid is constructed, and genes manB, manC, gmd, wcaG and zwf are overexpressed on a pETDuet-1 vector, so that the microbial cell can be used for preparing the microbial cell. In a 5L fermentation tank, after the final strain SL07 is used for final fermentation for 68 hours, the yield of 2 '-FL reaches 148g / L, and the production intensity reaches 2.18 g / L / h. The embodiment shows that the engineering strain for co-expressing the alpha-1, 2-fucosyltransferase from different sources can obviously improve the yield of the 2 '-FL.

The invention provides an L-arginine production strain and a construction method and application thereof, corynebacterium glutamicum AJC is modified by a metabolic engineering method to obtain the L-arginine production strain, the synthesis path of arginine is optimized, and a carbon source efficiently flows to arginine by means of promoter replacement, multi-copy and the like; by introducing an exogenous gene pyrABE949 *, the supply of a precursor substance carbamyl phosphate is enhanced; by introducing an exogenous gene pntAB, a new direction is provided for supply of coenzyme NADPH, and a large amount of reducing power is provided for synthesis of arginine; an exogenous gene fxpk is introduced, an NOG system is constructed, an acetyl coenzyme A pool is enriched, a large number of acetyl coenzyme A precursors are provided, and the engineering bacterium has the characteristics of high yield and stability and has good industrial application value in the aspect of fermentation production of L-arginine.

The invention relates to a strong promoter P7 suitable for streptomyces and application thereof, and relates to the field of genetic engineering and microbial metabolism engineering. The nucleotide sequence of the strong promoter P7 is as shown in SEQ ID No.1, and the strong promoter comprises a plasmid vector of the strong promoter; a host cell comprising the plasmid vector; the invention also discloses application of the strong promoter, the plasmid vector and the host cell in starting expression of a target gene. Compared with the prior art, the characterization of the strong promoter P7 provides an effective tool element for streptomyces strong promoter engineering and high-efficiency gene expression, and has important significance on streptomyces silent gene characterization, high-efficiency gene expression, metabolite synthesis, metabolic pathway reconstruction and the like. The strong promoter can be applied to common streptomyces type strains, and has important significance on high yield of important proteins including enzymes and important metabolites from actinomycetes.

The invention discloses pichia kudriavzevii for high yield of L-malic acid as well as a construction method and application thereof, and belongs to the technical field of biology, the pichia kudriavzevii is named as YM-023 and classified as Pichia kudriavzevii, the preservation number is CGMCC No. 35487, the preservation time is July 31, 2025, and the preservation number is CGMCC No. 35487. The preservation unit is China General Microbiological Culture Collection Center (CGMCC). The method comprises the following steps: starting from an acid-resistant yeast Kudriavzevii pichia pastoris strain YL-000 (the preservation number is CGMCC No.35680), carrying out metabolic engineering transformation to obtain a transformed strain, and ensuring that the transformed strain can realize efficient production of L-malic acid in a fermentation mode of adding a small amount of calcium carbonate neutralizer.

The invention belongs to the field of metabolic engineering, and discloses a genetically engineered bacterium for producing myo-inositol as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes escherichia coli as an original strain, and is obtained by performing the following gene editing on a genome of the escherichia coli: non-expressed lactose operon repressor protein, glucose phosphate dehydrogenase, acetokinase, glucose phosphate isomerase and glycerol repressor protein; and overexpressing inositol-1-phosphate synthase, inositol monophosphate, glucokinase, glucose permease and glycerol kinase. Compared with the prior art, the genetically engineered bacterium disclosed by the invention has remarkable advantages in the aspects of key enzyme expression rate, metabolism specificity, fermentation period and the like, and an efficient, stable, economical and feasible solution is provided for industrial production of myo-inositol.

The invention belongs to the technical field of molecular biology, and particularly relates to a DNA binding site of a fungus transcription factor AniJ and application. According to the invention, an echinocandin B gene cluster regulatory factor AniJ is taken as a research object, a ptH-SpRY-ACBE base editor and an sgRNA library covering an aniA promoter are utilized, 5-FOA screening is combined, and a binding motif RNGCTGAS of the AniJ in a core promoter region is excavated. The construction of an engineering strain containing a modified promoter (increasing the number of motifs) proves that the motifs can significantly enhance the expression of aniA and the yield of echinocandin B. And a novel molecular tool is provided for metabolic engineering of natural products of fungi.

The present disclosure provides a recombinant Escherichia coli strain modified by metabolic engineering means and a method for producing L-homoserine by using the same. The strain, designated as Escherichia coli having a strain number of 13-XA, is deposited in China General Microbiological Culture Collection Center (CGMCC) with an accession number of CGMCC No. 25099, dated Jun. 16, 2022. With respect to the chromosome DNA thereof, one or more genes associated with fatty acid metabolism are knocked out or attenuated, and / or a promoter is replaced for enhancement; one or more genes associated with the L-homoserine metabolic pathway are knocked out or attenuated, and / or one or more genes associated with the L-homoserine metabolic pathway are overexpressed or enhanced, and / or one or more genes associated with the L-homoserine metabolic pathway are mutated.

This invention relates to a combined module for increasing the yield of polyketide natural products from actinomycetes, and the construction and application of recombinant bacteria containing this module, belonging to the field of genetic engineering technology. To improve the yield of natural products such as polyketides and nonribosomal peptides, this invention utilizes a metabolic engineering strategy to combine and modify the CoASH synthetic pathway and the PPTase post-modification pathway, providing a combined module that can increase the yield of natural products from actinomycetes. This module consists of the CoASH cofactor synthesis module element SCoaA. R106A Composed of SCoaD and PPTase post-modification module element HPC3, recombinant vectors and recombinant bacteria containing this module were constructed. It was found that overexpression of this module can effectively increase the yield of natural products such as actinomycete polyketides, polyethers, and non-ribosomal peptides.

The invention belongs to the technical field of microbial fermentation, and relates to bacillus cereus capable of biologically converting limonene, the bacillus cereus is preserved in the China Center for Culture Collection, the classification name is Bacillus cereus GJ-02, the preservation number is CCTCC M 20251438, and the preservation date is June 23, 2025; according to the invention, the bacillus cereus strain capable of biologically converting limonene and having efficient conversion capability is separated and screened, a new strain resource is provided for microbial conversion of D-limonene, and a theoretical basis is provided for biosynthesis and metabolic engineering modification of terpenoids; therefore, the method has a wide application prospect in the field of microbial fermentation.

The invention provides a high-yield O-succinyl-L-homoserine engineering bacterium as well as a construction method and application of the high-yield O-succinyl-L-homoserine engineering bacterium. According to the invention, O-succinyl-L-homoserine genetically engineered bacteria with high yield of O-succinyl-L-homoserine are constructed by introducing O-succinyl-homoserine transferase from Vibrio cholera into a chassis strain in an exogenous manner and obtaining a gene mutant metAVc (T242A) of the O-succinyl-L-homoserine transferase, and further increasing the expression of the O-succinyl-L-homoserine transferase by adopting a Trc promoter. The engineering strain obtained through a systematic metabolic engineering modification strategy can realize effective accumulation of OSH, the shake flask yield of OSH reaches 18.8 g / L, the fed-batch fermentation yield of a 5L fermentation tank reaches 105.4 g / L, the sugar-acid conversion rate reaches 52.6%, and a foundation is laid for subsequent construction of high-yield OSH engineering bacteria.

The invention relates to the field of synthetic biology, metabolic engineering and fermentation engineering, and discloses a genetically engineered bacterium and application thereof in production of alpha-ketoglutaric acid.The genetically engineered bacterium comprises the steps that the genetically engineered strain with escherichia coli BW25113 or corynebacterium glutamicum ATCC13032 as a host is constructed; overexpression type isocitrate dehydrogenase genes are integrated through chromosomes, alpha-ketoglutarate dehydrogenase complex genes are knocked out, NADPH invertase is overexpressed through plasmids, and exogenous AKG transporter protein is introduced, so that metabolic flux directional regulation and control and product efflux enhancement are realized; meanwhile, a staged fermentation process is adopted and comprises dynamic gradient adjustment of inorganic phosphorus concentration and two-stage pH control. According to the method, the AKG yield of 78.2 g / L can be obtained under the 5L tank scale, the sugar acid conversion rate reaches 0.68, and the method has the advantages of being high in yield, low in byproduct and controllable in cost. The obtained AKG is suitable for a plurality of fields such as medical intermediates, functional foods and cell culture additives, and has wide industrial application prospects.

The invention belongs to the technical field of microbial fermentation, and relates to a nitrate reducing pseudomonas capable of biologically converting limonene, which is preserved in the China Center for Culture Collection, the classification name of which is Pseudomonas nitritiducens GJ-01, the preservation number of which is CCTCC M 20251439, and the preservation date of which is June 23, 2025, and the preservation number of which is CCTCC M 20251439. According to the invention, the nitrate reducing pseudomonas strain with efficient conversion capability and capable of biologically converting limonene is separated and screened, so that a new strain resource is provided for microbial conversion of D-limonene, and a theoretical basis is provided for biosynthesis and metabolic engineering modification of terpenoids; therefore, the method has a wide application prospect in the field of microbial fermentation.

The invention discloses radix tetrastigme flavone synthase and application thereof. The radix tetrastigme flavone synthase comprises an amino acid sequence as shown in SEQ ID No. 2. The key enzyme-flavone synthase gene (ThFNS) related to synthesis of apigenin in radix tetrastigme is cloned for the first time, in-vitro recombinant protein expression and enzymatic reaction system verification show that flavone synthase can catalyze naringenin to generate apigenin, and technical support is provided for follow-up metabolic engineering modification for an apigenin biosynthetic pathway.

The present invention belongs to the field of microbial metabolic engineering and synthetic biology technology applications, and particularly relates to a Pichia pastoris engineered strain that produces high sclareol production, as well as its construction method and application. A sclareol biosynthesis pathway is constructed in a host strain, and the intracellular mevalonate metabolic pathway and the central metabolic pathway are optimized, thereby obtaining an engineered bacterial strain A; the host strain is Pichia pastoris; or, metabolic regulatory factors are overexpressed or knocked out in the above-obtained engineered bacterial strain A, thereby obtaining an engineered bacterial strain B; or, in the above-obtained engineered bacterial strain B, a cell compartmentalization strategy is used to target the synthesis pathway to the peroxisome and optimize it, thereby obtaining an engineered bacterial strain C. The present invention provides a Pichia pastoris chassis cell for synthesizing the diterpenoid compound sclareol and an engineered strain for synthesizing sclareol; by introducing and optimizing the expression of the sclareol synthesis pathway, the sclareol yields in shake flask batch fermentation and bioreactor batch fed-batch fermentation reach 631.6 mg / L and 10.5 g / L, respectively.

The invention provides recombinant escherichia coli for synthesizing glucuronic acid by using glucose and a method for producing glucuronic acid by using the recombinant escherichia coli, and belongs to the field of microbial metabolic engineering. The engineering bacterium is obtained by taking escherichia coli as an original strain and knocking out a glucuronic acid isomerase coding gene uxaC, a 6-phosphofructokinase I coding gene pfkA, a 6-phosphofructokinase II coding gene pfkB and a glucose-6-phosphate dehydrogenase coding gene zwf. The engineering bacterium disclosed by the invention has a good application prospect in industrial production of glucuronic acid.

The invention belongs to the technical field of bioengineering, and discloses a genetic engineering strain for enhancing schizochytrium limacinum DHA oil production based on carbon flux guidance and fatty acid transport, a method and application thereof. The gene engineering strain is obtained by introducing an optimized coding gene ACLY and / or Fat1p coding gene related to guiding carbon flux and fatty acid transport into a schizochytrium limacinum body; wherein the gene sequence of the gene ACLY is SEQ ID No. 9, and the gene sequence of the gene Fat1p is SEQ ID No. 10. According to the invention, a schizochytrium limacinum engineering bacterium for enhancing DHA oil production based on carbon flux guidance and fatty acid transfer is constructed by means of metabolic engineering. The schizochytrium limacinum engineering bacterium is combined with fatty acid transfer based on carbon flux guiding, can be used for grease synthesis by guiding carbon flux, and can prevent grease from being degraded through fatty acid transfer.