30 results about "Phosphoketolase" patented technology

The present invention provides a bacterium which has an ability to produce a useful metabolite derived from acetyl-coenzyme A, such as L-glutamic acid, L-glutamine, L-proline, L-arginine, L-leucine, L-cysteine, succinate, and polyhydroxybutyrate, wherein said bacterium is modified so that activities of D-xylulose-5-phosphate phosphoketolase and / or fructose-6-phosphate phosphoketolase are enhanced. The present invention also provides a method for producing the useful metabolite using the bacterium.

Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.

The invention relates to a recombinant yeast, a construction method thereof and application thereof for producing tyrosol and a derivative. The recombinant yeast is acquired by introducing an exogenous fructose-6-phosphate phosphoketolase into an engineered yeast cell, and the engineered yeast cell is a yeast cell having a metabolic pathway for the synthesis of tyrosol via erythrose-4-phosphate and phosphoenolpyruvate. It is disclosed for the first time that in the process of expressing the fructose-6-phosphate phosphoketolase in the yeast, fructose-6-phosphate is catalyzed into erythrose-4- phosphate and acetyl phosphate while synthesizing 1,6-fructose diphosphate, xylulose-5-phosphate is catalyzed into glyceraldehydes-3-phosphate and acetyl phosphate, which alters the distribution of carbon metabolism in yeast, and enhances the synthesis of erythrose-4-phosphate, an important intermediate of tyrosol biosynthesis. The metabolic pathway of synthetic tyrosol is optimized, and the yieldof derivatives such as tyrosol and hydroxytyrosol is improved.

The invention provides for methods for the production of mevalonate, isoprene, isoprenoid precursor molecules, and / or isoprenoids in cells via the heterologous expression of phosphoketolase enzymes.

The invention provides a chassis system for ATP (Adenosine Triphosphate) regeneration and application. The chassis system for ATP regeneration comprises the following five enzymes: alpha-glucan phosphorylase, glucophosphomutase, phosphoglucose Isomerase, phosphoketolase and acetokinase. In addition, the chassis system can further comprise the following seven enzymes: transaldolase, transketolase,ribose-5-phosphate isomerase, ribulose-3 epimerase, triosephosphate isomerase, fructose-bisphosphate aldolase and fructose-1,6-bisphosphatase. The chassis system can be coupled to an enzymic catalyticreaction needing ATP, utilizes starch or maltodextrin as energy, is not added with any coenzyme, can efficiently regenerate the ATP at low cost through a one-pot reaction and is an economical, highlyefficient, stable and sustainable ATP regeneration system.

Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.

Provided herein are compositions and methods for improved production of acetyl-CoA and acetyl-CoA derived compounds in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding a phosphoketolase (PK), and a functional disruption of an endogenous enzyme that converts acetyl phosphate to acetate. In some embodiments, the host cell further comprises a heterologous nucleotide sequence encoding a phosphotransacetylase (PTA). In some embodiments, the enzyme that converts acetyl phosphate to acetate is a glycerol-1-phosphatase. In some embodiments, the glycerol-1-phosphatase is GPP1 / RHR2. In some embodiments, the glycerol-1-phosphatase is GPP2 / HOR2. The compositions and methods described herein provide an efficient route for the heterologous production of acetyl-CoA-derived compounds, including but not limited to, isoprenoids, polyketides, and fatty acids.

The invention discloses a genetically engineered bacterium capable of increasing the yield of beta-carotene and application thereof, and the genetically engineered bacterium contains a beta-carotene synthetic pathway related gene, a phosphoketolase pathway related gene and a lipid droplet synthetic pathway related gene. A beta-carotene synthetic pathway is covered; a phosphoketolase pathway is increased, the carbon loss is reduced, and more precursor substance acetyl coenzyme A for synthesizing beta-carotene is generated from a carbon source; the supply of a reducing power substance NADPH and an energy substance ATP is increased; and a lipid droplet synthesis pathway is also increased, and the storage of beta-carotene can be increased by increasing the content and volume of intracellular lipid droplets. The highest yield of the beta-carotene of the genetically engineered bacterium reaches 7.8g / L, and the yield of the beta-carotene is remarkably improved.

Substance productivity is improved by introducing a metabolic pathway for synthesis of acetyl-CoA or acetic acid from glucose-6-phosphate into yeast. Acetic acid productivity, acetyl-CoA productivity, and productivity of a substance made from acetyl-CoA-derived are improved by attenuating genes involved in the glycolytic system of yeast and introducing a phosphoketolase gene into the yeast.

Substance productivity is improved by introducing a metabolic pathway for synthesis of acetyl-CoA or acetic acid from glucose-6-phosphate into yeast. Acetic acid productivity, acetyl-CoA productivity, and productivity of a substance made from acetyl-CoA-derived are improved by attenuating genes involved in the glycolytic system of yeast and introducing a phosphoketolase gene into the yeast.

A recombinant yeast is constructed by introducing an expressed gene of exogenous Fructose-6-phosphate phosphoketolase into a modified yeast cell, and the modified yeast cell is a yeast cell with a metabolic pathway for synthesizing tyrosol via Erythrose-4-phosphate and phosphoenolpyruvate. The present invention discloses for the first time that in the process of expressing Fructose-6-phosphate phosphoketolase in a yeast, Fructose-6-phosphate is synthesized into beta-D-Fructose 1,6-bisphosphate and also catalyzed into Erythrose-4-phosphate and Acetyl-phosphate, and Xylulose-5-phosphate is catalyzed into Glyceraldehydes-3-phosphate and Acetyl-phosphate, which change the metabolic flux distribution of carbon in the yeast, enhance the synthesis of Erythrose-4-phosphate as an important intermediate for the biosynthesis of tyrosol, optimize the metabolic pathway for synthesizing tyrosol, and increase the yields of tyrosol and its derivatives such as hydroxytyrosol.

The invention provides phosphoketolases for improved production of acetyl coenzyme a-derived metabolites, isoprene, isoprenoid precursors, and isoprenoids. This present invention relates to cultured recombinant cells comprising a heterologous phosphoketolase (PKL) polypeptide that are capable of increased production of acetyl coenzyme A-derived metabolites, as well as methods for producing and using the same. In some embodiments, the recombinant cells further comprise one or more mevalonate (MVA) pathway polypeptides for the production of isoprenoid precursors, isoprene and isoprenoids.

The invention discloses a phosphoketolase with improved activity and its use in the production of metabolites. The protein provided by the present invention is a mutant protein, which is a protein obtained by subjecting phosphoketolase to any one or more of the following mutations: the second position is mutated from T to A; the sixth position is mutated from I to T; the 14th position The position is mutated from N to D; (the 20th position is mutated from E to D; the 120th position is mutated from T to A; the 231st position is mutated from E to K; the 260th position is mutated from H to Y; the 342nd position is mutated from E It is K; the 397th position is mutated from K to R; the 676th position is mutated from D to G; the 785th position is mutated from F to L; the 801st position is mutated from W to R. The present invention also protects the mutant protein when preparing metabolites Application in. Compared with the existing phosphoketolase, the phosphoketolase activity of the mutant protein provided by the invention is significantly increased, thereby significantly increasing the yield of target metabolites.

Described are compositions and methods relating to yeast expressing glycerol dehydrogenase and dihydroxyacetone kinase polypeptides in combination with an exogenous phosphoketolase pathway, as well asto bifunctional glycerol dehydrogenase-dihydroxyacetone kinase fusion polypeptides, and their various and combined uses in starch hydrolysis processes for alcohol production.

Provided is a lactic acid bacterium capable of homolactic fermentation using a pentose as a substrate, the lactic acid bacterium utilizing a pentose, and in which a phosphoketolase pathway is blocked and a pentose phosphate pathway is activated. Also provided is a method for producing lactic acid from a pentose using the lactic acid bacterium and a method for preparing the lactic acid bacterium.

The invention discloses a method for increasing the cell phloroglucinol synthesis yield and application, and belongs to the technical field of genetic engineering. According to the method provided by the invention, a recombined cell co-expresses a phosphoketolase fxpk gene, a fructose 1,6-biphosphatase fbp gene and a poly-ketene anhydride synthetase phld gene and also expresses any one or two of a multiple-resistance-factor MarA gene and an acetyl-coenzyme a carboxylase ACCase gene. According to the method provided by the invention, a new phloroglucinol synthesis way is built in the recombined cell, so that the yield of phloroglucinol is greatly increased; as for a contrast group, the phloroglucinol yield is increased by 68.9 percent; meanwhile, by the method, emission of carbon dioxide in a phloroglucinol synthesis process is also reduced; the method has high environmental benefits.

The invention discloses a high-yielding β-ionone genetically engineered bacterium and its construction method and application, belonging to the field of microbial fermentation. The method is to successfully construct high-yield β-ionone genetically engineered bacteria by strengthening the MVA pathway and introducing the phosphoketolase pathway to increase the supply of acetyl-CoA in the cytoplasm and the metabolic flow of the terpenoid synthesis pathway. The genetically engineered bacteria obtained according to the method of the present invention can produce β-ionone under conventional fermentation conditions, the shake flask fermentation output can reach about 358mg / L, and the 3L fermenter output can reach about 0.98g / L, which is close to industrialization Level. The invention can use simple culture medium to ferment and produce spice β-ionone, can realize one-time high-efficiency transformation and integration of multiple genes, can significantly shorten the construction time of engineering bacteria, and the obtained engineering bacteria can use simple carbon sources such as glucose and glycerin to produce spice β-ionone Fermentative production of ionone has good industrial application prospects.