242 results about "Acetyl coenzyme" patented technology

The present disclosure provides a recombinant yeast host cell having a plurality of native and / or heterologous enzymes that function in an engineered metabolic pathway to convert fructose-6-phosphate to acetyl-coenzyme A. The recombinant yeast host cell can be used, for example, in a process for making acetyl-coenzyme A, acetone and / or isopropanol. The plurality of native and / or heterologous enzymes is activated, upregulated, or overexpressed. The plurality of native and / or heterologous enzymes comprises a phosphoketolase; and optionally an acetate kinase; and / or a phosphotransacetylase. The recombinant yeast host cell comprises at least one of: at least two copies of a heterologous nucleic acid molecule encoding the phosphoketolase; a native and / or heterologous enzyme that functions in an engineered non-oxidative pentose phosphate pathway, wherein the native and / or heterologous enzyme is activated, upregulated, or overexpressed; and / or a native and / or heterologous protein that functions in an engineered metabolic pathway to convert pantothenate into acetyl-coenzyme A, wherein the native and / or heterologous protein is activated, upregulated or overexpressed and comprises at least one of: FEN2 or CAB1.

The invention relates to the technical field of genetic engineering, and discloses a genetically engineered bacterium for producing L-arginine as well as a construction method and application of the genetically engineered bacterium. Escherichia coli is taken as a starting strain, and the following treatment is performed: overexpressing genes gdhA and gdh of glutamate dehydrogenase, promoting generation of glutamic acid, overexpressing and coding arginine transporter gene argO to enhance extracellular transport of arginine, introducing exogenous argJ to replace a linear synthesis pathway in large intestines, improving the utilization rate of acetyl coenzyme A and accelerating the synthesis efficiency of arginine; a glutamine synthetase gene glnA is over-expressed, and glutamine synthesis is enhanced; the pckA is overexpressed to promote ATP synthesis; meanwhile, speA and adiA genes for coding the arginine decarboxylase are deleted, so that further synthesis of putrescine from L-arginine is inhibited, and the L-arginine accumulation capability of the strain is improved. Through the combined action of the treatment, a genetically engineered bacterium with extremely high L-arginine production yield is finally obtained, and the shake flask fermentation yield can reach 13.7 g / L.

The invention belongs to the field of biosynthesis, and in particular relates to LBD (Levandanenediol Pyrophosphate) synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, the sclareol biosynthesis yield is generally low, LSLPPs mutant containing at least one mutation of D730L, D374L, N674L or G379M is obtained by performing mutation and optimization on LSLPPs pyrophosphate synthase, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 26.11 g / L and is greatly improved compared with the prior art.

The invention discloses a recombinant strain for producing squalene as well as a construction method and application of the recombinant strain. The construction method comprises the following steps: heterologously expressing cDNA (complementary deoxyribonucleic acid) of each gene in a squalene synthesis route from filamentous fungi in yeast; the squalene synthetic pathway genes comprise a mevalonic acid-5-phosphate kinase gene, a squalene synthase gene, an acetoacetyl coenzyme A thiolase gene, a mevalonic acid kinase gene, a hydroxymethyl glutaryl coenzyme a synthetase gene, a mevalonic acid pyrophosphate decarboxylase gene, a farnesyl pyrophosphate synthetase gene and isopentenyl pyrophosphate. A dimethallylpyrophosphate isomerase gene and a truncated hydroxymethyl glutaryl coenzyme a reductase gene; the culture medium is characterized in that the filamentous fungi are Saroocladium oryzae or Stibella sp. 1056, and the culture medium is a culture medium. The excavated squalene synthesis route can complete efficient de novo synthesis of squalene, and the squalene yield is increased.

The invention belongs to the field of biosynthesis, and particularly relates to sclareol synthase and engineered yeast for producing sclareol. In order to solve the problem that in the prior art, sclareol biosynthesis yield is generally low, sclareol synthase is mutated and optimized, an SsScs mutant containing at least one mutation of Y491F, G307L or N269F is obtained, and the mutant is used for sclareol synthesis. Meanwhile, in order to solve the problem of genetic instability caused by free plasmid expression of part of saccharomyces cerevisiae strains, a sclareol synthesis pathway is constructed in a yeast genome, and meanwhile, an acetyl coenzyme A synthesis pathway, an MVA pathway and a sclareol synthesis pathway are enhanced; chassis bacteria for synthesizing sclareol are modified in a manner of knocking out part of transcription inhibition factors, and the yield of a 5L fermentation tank reaches 21.13 g / L and is greatly improved compared with the prior art.

The invention discloses a pichia pastoris engineering bacterium for synthesizing bakuchiol from de novo by converting methanol as well as construction and application of the pichia pastoris engineering bacterium. A heterologous p-coumaric acid synthesis pathway is introduced to neutral sites of different chromosomes of pichia pastoris, a high-yield p-coumaric acid strain is obtained through gene knockout, overexpression or heterologous expression of tyrosine and phenylalanine synthesis pathway key genes of a shikimic acid pathway, a heterologous bakuchiol biosynthesis pathway is introduced on the basis, and a high-yield p-coumaric acid strain is obtained. After overexpression of bakuchiol synthetase, endogenous overexpression or heterologous expression of MVA pathway key genes and exogenous acetyl coenzyme A supply pathway genes, methanol concentration optimization and bacterial strain His4 gene supplementation, the yield of bakuchiol is effectively increased to 91.2 mg / L and is increased by 59.8 times compared with that of an initial bacterial strain, and the yield of bakuchiol in a 15L fermentation tank reaches 692.8 mg / L. The method has the characteristics of high conversion efficiency, low production cost, convenience in preparation, wide industrial application prospect and the like.

The invention provides a euphorbia lathyris acyltransferase gene ElBAHD16 and application of a coding protein thereof. The gene is obtained by cloning from euphorbia lathyris for the first time. The invention also provides a recombinant vector and an engineering bacterium containing the gene. Cinnamoyl coenzyme A, benzoyl coenzyme A or acetyl coenzyme A can be used as an acyl donor, and 7-hydroxy euphorbia lathyris diterpenoid alcohol is used as a substrate to generate a plurality of types of euphorbia lathyris diterpenoid esters through biological catalysis. The content of diterpenoid ester substances in plants such as euphorbia lathyris can be increased through a genetic engineering technology, and the method has a good application prospect.

The invention discloses a hybrid central metabolic network chassis cell with both carbon atom economy and energy generation pathway as well as a construction method and application of the hybrid central metabolic network chassis cell, and belongs to the technical field of genetic engineering and microorganisms. The Escherichia coli central metabolic network is reconstructed, so that the Escherichia coli central metabolic network has a carbon sequestration module and an energy generation module at the same time. The carbon sequestration module continues to fix CO2 while avoiding CO2 release caused by pyruvic acid decarboxylation, the highest carbon yield can reach 133%, the carbon yield exceeds that of an NOG pathway reported only from the perspective of no CO2 release, and the carbon yield is two times that of a natural glycolysis pathway. And the direct product of the constructed mixed type central metabolic network is acetyl coenzyme A, and the acetyl coenzyme A contains an energy generation module, so that the chassis cell provided by the invention can meet the synthesis requirements of acetyl coenzyme A derivatives with high or low required reducing power, and has universality.

The invention belongs to the technical field of genetic engineering, and discloses a schizochytrium limacinum genetic engineering strain for enhancing supply of acetyl coenzyme A and efficiently producing astaxanthin, a method and application of the schizochytrium limacinum genetic engineering strain. By heterologous expression of a beta-carotene ketolase CrtO gene and a beta-carotene hydroxylase CrtZ gene from haematococcus pluvialis, synthesis of astaxanthin from beta-carotene in schizochytrium limacinum is promoted, so that the content of astaxanthin in schizochytrium limacinum HX-308 is increased. According to the invention, the endogenous AACT gene and heterologous CrtO, CrtZ, PK and PTA genes of schizochytrium limacinum are integrated into an original strain through homologous recombination, and the obtained strain still keeps genetic stability after multiple subculture, has the capability of efficiently synthesizing astaxanthin, effectively improves the yield of astaxanthin, and lays a foundation for large-scale industrial production.

The invention discloses a rice ACCase mutant protein and a preparation method thereof. The amino acid sequence of the ACCase mutant gene encoding protein has the following mutations: the 2052nd site corresponding to the amino acid sequence of wild rice ACCase is mutated, isoleucine (I) is changed into asparagine (N), or isoleucine (I) is changed into glutamine (Q), or isoleucine (I) is changed into cysteine (C), or isoleucine (I) is changed into cysteine (C), or isoleucine (I) is changed into cysteine (C), or isoleucine (I) is changed into cysteine (C), or isoleucine (I) is changed into cysteine (C). And the plant can have extremely strong resistance (tolerance) to acetyl-coenzyme A carboxylase inhibitor herbicides. After the rice containing the rice ACCase mutant protein disclosed by the invention is applied with an acetyl-coenzyme A carboxylase inhibitor herbicide with 8 times of recommended use concentration, plants still grow normally, and the tolerance of the herbicide is greatly enhanced, so that the rice ACCase mutant protein can be widely applied to breeding of herbicide-resistant plants.

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 present invention discloses the application of ginseng exosomes in the preparation of nanomedicines for treating diabetic ulcer vascular lesions, and relates to the field of biomedicine. The present invention verifies through experiments that ginseng exosomes can reprogram the glycolysis process of vascular endothelial cells under high sugar conditions, and inhibit the expression of acetyl coenzyme A in cells, reduce the production of reactive oxygen species, and then promote the expression of nitric oxide synthase, thereby reducing or reversing cell apoptosis induced by high sugar. Ginseng exosomes are used as nanomedicines to prevent, treat or improve diabetic ulcer microangiopathy. The ginseng exosomes in the present invention have a simple preparation process, low cytotoxicity, high delivery efficiency, and good stability. They can be used as a new type of nanomedicine for the prevention, treatment or improvement of diseases related to diabetic vascular lesions.

The invention belongs to the technical field of bioengineering, and particularly relates to kluyveromyces marxianus engineering bacteria for synthesizing eicosapentaenoic acid and application of the kluyveromyces marxianus engineering bacteria. According to the engineering bacterium, pyruvate decarboxylase PDC1 genes, alcohol dehydrogenase ADH1 genes and alcohol dehydrogenase ADH2 genes are sequentially knocked out from a kluyveromyces marxianus FIM1 delta ura3 strain; meanwhile, a citrate lyase ACL gene, a pyruvate oxidase PO gene and a phosphate transacetylase PTA gene are inserted into the loci respectively, so that synthesis of acetyl coenzyme A is enhanced; then a plurality of genes are sequentially inserted into a Kluyveromyces marxianus genome, and then a vector of a delta 8 desaturase delta 8D gene is transferred into the Kluyveromyces marxianus genome. The invention also provides a method for efficiently preparing the EPA, namely the Kluyveromyces marxianus engineering strain is used as a fermentation strain, glucose, a yeast extract, peptone and other components are used as a culture medium, and the yield of the EPA produced by culturing and fermenting reaches 8.71 mg / g of dry cell weight.

Provided are a brewer's yeast having low ethanol synthesis and high acetyl-CoA synthesis, and a use thereof, belonging to the technical field of biology. In order to reduce the accumulation of ethanol, the main byproduct of brewer's yeast, and increase synthesis of acetyl-CoA, MTH1, MED2 and HXT2 transcription factors are modified, an optimal transcription factor combination for reducing ethanol synthesis is obtained by screening, and then heterologous acetyl-CoA synthesis pathways of different synthesis pathways integrated in the brewer's yeast genome are compared, and a recombinant brewer's yeast strain is provided. The strain can grow in a sterile culture medium using glucose as a carbon source, creating a platform for modifying brewer's yeast by metabolic engineering, to synthesize high-value compounds; in addition, the construction method is simple and easy to use, and has good application prospects.

The invention discloses yarrowia lipolytica with high yield of beta-carotene as well as a construction method and application of the yarrowia lipolytica. According to the yarrowia lipolytica engineering bacterium, geranyl diphosphate synthase gene xdGPS, phytoene dehydrogenase gene CarB, phytoene cyclization / phytoene synthesis bifunctional enzyme mutant gene GarRPY27R, acetyl-coenzyme A synthetase mutant gene ACSL641P and pyruvate ferredoxin oxidoreductase gene nifJ are integrated and expressed on a chromosome, and the yarrowia lipolytica engineering bacterium is obtained. Meanwhile, a 3-hydroxy-3-methylglutaryl CoA reductase gene HMGR (3-hydroxy-3-methylglutaryl CoA reductase) derived from the yarrowia lipolytica strain is subjected to overexpression; the yield of the beta-carotene is increased to 13.186 g / L, and the production efficiency is 0.14 g / L / h.

A method for preparing glycine by using threonine relates to a fermentation process in which threonine is decomposed into glycine and acetaldehyde by aldolase. Glycine and acetyl coenzyme A can be produced in a fermentation process, in which acetaldehyde is reduced into acetyl coenzyme A or an acetyl coenzyme A derivative by acetylating acetaldehyde dehydrogenase; or threonine is dehydrogenated by threonine dehydrogenase to obtain 2-amino-3-ketobutyric acid, which is then ligated by 2-amino-3-ketobutyrate CoAligase to obtain acetyl coenzyme A. Coenzyme A can be converted into an acetyl coenzyme A derivative under different fermentation conditions.

The invention discloses a method for biosynthesizing dicarbon organic acid by using CO2, and belongs to the technical field of bioengineering. A path for synthesizing glyoxylic acid and glycollic acid by taking CO2 as a substrate is designed, and a first reaction module for converting CO2 and acetyl coenzyme A into pyruvic acid and a second reaction module for converting pyruvic acid into malic acid are obtained through thermodynamic evaluation and screening of pathase. The third reaction module is used for converting malic acid into acetyl coenzyme A and glyoxylic acid, the fourth reaction module is used for converting glyoxylic acid into glycollic acid, and PSP circulation is carried out on the four reaction modules. Through an experimental method of a one-pot two-step method, 2.78 g / L of glyoxylic acid can be realized after 11 hours of catalytic reaction in total; and 1.78 g / L of glycollic acid. The invention successfully realizes a cell-free biosynthesis method for synthesizing glyoxylic acid and glycollic acid by taking CO2 as a substrate.

The present disclosure relates to multi-enzyme pathways for the in vitro cell-free biosynthesis of the cofactor, coenzyme A ("CoA") from lower-cost substrates, such as D-pantothenate, L-cysteine, and ATP, and the use of these pathways in CoA-dependent cell-free systems for the biobased production of chemicals, such as acetyl-CoA, hexanoyl-CoA, malonyl-CoA, and cannabigerolic acid (CBGA).

The application provides a recombinant Yarrowia lipolytica capable of producing palmitoleic acid at a high yield, a construction method and application thereof, and belongs to the technical field of bioengineering. Yarrowia lipolytica The recombinant Yarrowia lipolytica is obtained by knocking out peroxisome biogenesis factor 10 and triacylglycerol lipase 4 in the genome of Yarrowia lipolytica and inserting an acetyl-CoA carboxylase, a Δ9 desaturase, an acetyl-CoA diacylglycerol acyltransferase, a glycerol-3-phosphate acyltransferase, a lysophosphatidylcholine acyltransferase and a phosphatidylcholine:diacylglycerol acyltransferase gene expression cassette. Experiment proves that the recombinant Yarrowia lipolytica can efficiently ferment and produce palmitoleic acid, and realizes efficient synthesis of the natural product palmitoleic acid from plants in Yarrowia lipolytica.

Described is a recombinant organism or microorganism which is capable of enzymatically converting acetyl-CoA into isobutene, (A) wherein in said organism or microorganism: (i) acetyl-CoA is enzymatically converted into acetoacetyl-CoA, (ii) acetoacetyl-CoA is enzymatically converted into 3-hydroxy-3-methylglutaryl-CoA, (iii) 3-hydroxy-3-methylglutaryl-CoA is enzymatically converted into 3-methylglutaconyl-CoA, (iv) 3-methylglutaconyl-CoA is enzymatically converted into 3-methylcrotonyl-CoA, and (v) wherein said 3-methylcrotonyl-CoA is converted into isobutene by: (a) enzymatically converting 3-methylcrotonyl-CoA into 3-methylcrotonic acid which is then further enzymatically converted into said isobutene; or (b) enzymatically converting 3-methylcrotonyl-CoA into 3-hydroxy-3-methylbutyryl-CoA which is then further enzymatically converted into 3-hydroxy-3-methylbutyric acid which is then further enzymatically converted into 3-phosphonoxy-3-methylbutyric acid which is then further enzymatically converted into said isobutene; (B) wherein said recombinant organism or microorganism has an increased pool of coenzyme A (CoA) over the organism or microorganism from which it is derived due to: (i) an increased uptake of pantothenate; and / or (ii) an increased conversion of pantothenate into CoA. Moreover, described is the use of such a recombinant organism or microorganism for the production of isobutene. Further, described is a method for the production of isobutene by culturing such a recombinant organism or microorganism in a suitable culture medium under suitable conditions.

The invention discloses an engineering strain for producing glycine. The engineering strain is obtained by modifying the following steps: (i) respectively introducing an exogenous acetyl coenzyme A synthetase gene acs, an isocitrate lyase gene aceA and an isocitrate dehydrogenase kinase gene aceK; (ii) introducing a glutamic acid glyoxylate transaminase gene cgaT from quinoa; and (iii) respectively knocking out a DNA (Deoxyribose Nucleic Acid) binding transcription inhibition factor gene iclR, a glycine dehydrogenase gene gcvP, an alanine transaminase gene alaA, a malate dehydrogenase gene maeA and a malate synthetase G gene glcB. According to the invention, an engineering strain capable of efficiently synthesizing glycine is constructed by transforming a glycine biosynthesis metabolic pathway, and the strain can produce glycine by fermentation with glucose as a carbon source and has a relatively high conversion rate.

The present invention provides a mutant acetyl-CoA carboxylase (ACC) protein, a nucleic acid and an application thereof, and in particular, relates to a mutant acetyl-CoA carboxylase (ACC) protein, a nucleic acid and an application thereof in plant breeding. In particular, the present invention provides a mutant acetyl-CoA carboxylase (ACC) protein, and the mutant acetyl-CoA carboxylase (ACC) protein is compared with the parent acetyl-CoA carboxylase (ACC) protein in the region corresponding to SEQ ID Mutations occur at any one or several of the following amino acid positions in the amino acid sequence shown in No. 1: 2125, 2097, 2139, 2194, 2186, 2273, 2168, 1975, 1954, 1864, 2211, 2187, 2123, and 2126; plants with the acetyl-CoA carboxylase (ACC) mutation have high herbicide resistance and have very broad application prospects in breeding herbicide-resistant plants.

The present disclosure relates to the use of a switch for the production of heterologous non-catabolic compounds in microbial host cells. In one aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured under aerobic conditions followed by microaerobic conditions, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions. In another aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured in the presence of maltose followed by the reduction or absence of maltose, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions.

The present invention relates to tomato plants having improved insect resistance, more particularly whitefly resistance or mite resistance, wherein the plants comprise an ASAT3 gene encoding an acetyl-CoA-dependent acyltransferase and an AP2e gene encoding an APETALA2 ethylene response transcription factor. The invention also relates to a method for providing tomato plants with improved insect resistance, and the use of the ASAT3 gene in combination with the AP2e gene for providing insect resistant tomato plants.

This invention relates to a recombinant Halomonas strain that produces polyhydroxyalkanoates (PHA) by overexpressing endogenous enzymes encoding ADP-dependent acetyl-CoA synthase (ADP-ACS). acd This gene not only enhances the recombinant bacterium's tolerance to acetic acid but also enables it to efficiently convert acetic acid or acetate substrates into acetyl-CoA, thereby participating in the tricarboxylic acid cycle (TCA cycle) and the synthesis of PHA. The present invention also relates to a method for producing PHA using the aforementioned recombinant Halomonas bacterium employing a pH-stat strategy.

The present disclosure describes a recombinant yeast host cell having a first engineered metabolic pathway to convert fructose-6-phosphate (F-6-P) into acetyl-coenzyme A. The first engineered metabolic pathway comprises: a genetic modification for expressing a heterologous phosphoketolase capable of converting fructose-6-phosphate into erythrose-4-phosphate, and at least one genetic modification for decreasing native 6- phosphofructo-2-kinase activity classified under Enzyme Commission No. 2.7.1.105. Optionally, the recombinant yeast host cell comprises a second engineered metabolic pathway for convert acetyl-coA into a fermentation product. The present disclosure further describes a process for making a fermentation product. The process comprises contacting the recombinant yeast host cell described herein with a carbohydrate source under a condition allowing the conversion of at least a part of the carbohydrate into a fermentation product.

The invention relates to the technical field of substance detection, and particularly discloses an enzyme activity determination method of acetyl-CoA carboxylase, a kit and application. The invention provides an enzyme activity determination method of acetyl coenzyme A carboxylase. The detection reaction process is as follows: # imgabs0 #. The detection method provided by the invention can realize rapid and accurate detection of ACC enzyme activity, can perform efficient ACC determination in research and application, and has a wide application prospect.

The invention provides non-naturally occurring microbial organisms containing enzymatic pathways and / or metabolic modifications for enhancing carbon flux through acetyl-CoA. In some embodiments, the microbial organisms of the invention having such pathways also include pathways for generating reducing equivalents, formaldehyde fixation and / or formate assimilation. The enhanced carbon flux through acetyl-CoA, in combination with pathways for generating reducing equivalents, formaldehyde fixation and / or formate assimilation can, in some embodiments, be used for production of a bioderived compound. Accordingly, in some embodiments, the microbial organisms of the invention can include a pathway capable of producing a bioderived compound of the invention. The invention still further provides a bioderived compound produced by a microbial organism of the invention, culture medium having the bioderived compound of the invention, compositions having the bioderived compound of the invention, a biobased product comprising the bioderived compound of the invention, and a process for producing a bioderived compound of the invention.

The invention discloses a genetically engineered bacterium for synthesizing N-acetyl blue as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium is prepared by knocking out a lactic dehydrogenase gene ldh and inserting a phosphoketolase gene xfpK on a knockout site, knocking out a pyruvate oxidase gene poxB and inserting a phosphate transacetylase gene pta on a knockout site of a microorganism for producing N-acetyl blue, and knocking out an aconitase inhibition factor gene acnR, inserting a glutamate dehydrogenase gene gdhA on a knockout site, knocking out a glutamine synthetase adenylation transferase gene glnE, and inserting an alpha-ketoglutarate dehydrogenase inhibition factor gene odhI on a knockout site to obtain the strain. According to the method, the supply of the precursor acetyl coenzyme A is enhanced by introducing a non-oxidative glycolysis pathway, the pH dynamic regulation system balances the metabolic flux, and the double breakthrough of the yield and the carbon recovery rate of the N-acetyl blue is realized by combining a two-stage fermentation process.