12 results about "Pyruvic dehydrogenase" patented technology

The application belongs to the field of biological analysis, and provides a method for precise metabolic flow analysis of TCA cycle based on isotope tracing technology combined with high-resolution mass spectrometry. By optimizing mass spectrometry parameters and liquid chromatography method, one injection within 6 min is realized, and high-throughput, high-sensitivity and high-resolution detection is realized on 11 biomarkers related to TCA cycle metabolic pathways. At the same time, the application innovatively uses LC-MS / MS to decipher 13 C the stepwise position-specific transfer from glucose to subsequent metabolites by glycolysis and TCA cycle, thereby calculating 13 C the integrated steady-state analysis of key metabolic rates of position-specific transfer from sequential precursors to their products (pyruvate dehydrogenase, beta-oxidation, pyruvate carboxylase, isocitrate dehydrogenase and pyruvate cycle). This technology has wide applicability and can potentially characterize mitochondrial metabolism of any tissue or cell.

The application discloses a kind of modified bacteria for producing L-alanine, compared with wild type bacteria, the genome is integrated with pyruvate dehydrogenase alaD coding gene of bacillus stearothermophilus origin and the coding gene of mutant alaE (A48S / A149D) of alanine export protein alaE, and the expression of gene ldhA, mgsA, pflB, adhE, ackA and frdB in chassis bacterial genome is interrupted.The application also discloses a kind of method for quickly constructing recombinant escherichia coli for producing L-alanine, the method is suitable for multiple escherichia coli subspecies including MG1655, BL21 and ATCC8739, has universality and popularization and application value.

The invention discloses an artificial synthesis method and application of malonyl coenzyme A. An artificial synthesis route for synthesizing malonyl-CoA by taking beta-alanine (beta-ala) as a precursor is constructed by heterologous expression of aminotransferase and malonyl-CoA reductase: firstly, under the catalysis of transaminase, amino of beta-ala is transferred to alpha-keto acid (such as pyruvic acid, oxaloacetic acid or alpha-ketoglutaric acid and the like), and then the malonyl-CoA is obtained; an intermediate product 3-oxopropionic acid and corresponding amino acid are formed; and the malonyl-CoA is generated from the 3-oxopropionic acid under the action of the malonyl-CoA reductase. According to the method, the defects existing in a natural malonyl-CoA synthesis route, such as low carbon utilization rate, energy substance ATP consumption, greenhouse gas CO2 release, pyruvate dehydrogenase (PDH) of route enzymes and acetyl-CoA carboxylase (ACC) which are strictly regulated and controlled, are overcome, and high yield of products taking malonyl-CoA as a precursor, including light flavomycin, octanoic acid, phloroglucinol, pentadecheptaene, natamycin, spinosad and the like, is realized.

The invention discloses a construction method of escherichia coli for efficiently synthesizing glycollic acid, and belongs to the technical field of biochemical engineering. According to the invention, citric acid synthase gltA and aconitic acid synthase acnA and acnB are overexpressed in escherichia coli cells, pyruvate dehydrogenase transcription inhibition factor pdhR is knocked out, transhydrogenase pntA and pntB are overexpressed, and transhydrogenase stA is knocked out, so that the constructed recombinant escherichia coli can produce glycollic acid by using glucose in a short time. The reaction cost is lower, the yield is higher, the concentration of glycollic acid produced after fermentation in a 5L fermentation tank for 48 hours is 81 g / L, the conversion rate is 0.62 g / g, and the production intensity is 1.69 g / L / h.

The invention discloses a construction method and application of an engineering bacterium for increasing the yield of alpha-ketoglutaric acid. Wild E.coli.W3110 serves as a chassis strain, lactic dehydrogenase ldhA, pyruvate dehydrogenase poxB and formic acid C-acetyltransferase pflB are knocked out firstly, and the flow direction of pyruvic acid to lactic acid, acetic acid and formic acid is blocked; meanwhile, the promoter PBBaJ23109 is used for replacing a natural promoter of the sucA gene, so that alpha-ketoglutaric acid dehydrogenase is weakened, and the decomposition way of alpha-ketoglutaric acid is weakened; the phosphoenolpyruvate carboxylase ppc is overexpressed, and the flow direction of phosphoenolpyruvate to oxaloacetic acid is enhanced; isocitrate dehydrogenase icd from escherichia coli is overexpressed, so that synthesis of a product is promoted; besides, pyruvate dehydrogenase derived from saccharomyces cerevisiae is introduced and coded by genes PDB1, PDA1 and LAT1, so that an important precursor, namely pyruvic acid, is promoted to flow to acetyl coenzyme A; the engineering strain with high yield of alpha-ketoglutaric acid is finally obtained, the yield of 75.1 g / L can be achieved in a 5L fermentation tank through a fed-batch fermentation method for 40 hours, and the conversion rate is 39.6%.

The present invention relates to recombinant Modified Vaccina Virus Ankara (MVA) encoding Epstein-Barr virus (EBV) antigens, wherein surface glycoprotein 350 (EBV gp350) and glycoprotein gH (EBV gH) are fused to subunits of self-assembling multimeric protein particle PdhC (acetyltransferase of pyruvate dehydrogenase (PDH) complex) or DPS (DNA binding protein from starved cells).

This application relates to a microorganism in which the activity of pyruvate dehydrogenase complex subunit E1 is increased, and to a method for producing O-acetylhomoserine or its derivatives using the same.

This application relates to a microorganism in which the activity of pyruvate dehydrogenase complex dihydrolipoyllysine residue acetyltransferase is improved, and to a method for producing L-amino acids using the same.

This invention relates to the field of agricultural biotechnology, specifically to a method and strain for constructing an engineered strain of *Hookworm Copper-Gropping Bacteria* capable of producing methylmalic acid using both organic and inorganic carbon sources. This application involves knocking out the restriction endonuclease encoding gene of the type I restriction modification system in the genome of wild-type *Hookworm Copper-Gropping Bacteria* H16. 、 Promoters adapted for methylmalate synthase expression in *Hypoxonometra* were screened, and methylmalate synthases with high catalytic activity in *Hypoxonometra* were obtained, capable of effectively catalyzing methylmalate synthesis in *Hypoxonometra*. A methylmalate transporter protein that promotes methylmalate excretion was also screened, as well as a gene encoding the *Hypoxonometra* endogenous pyruvate dehydrogenase complex subunit that promotes methylmalate synthesis when sodium gluconate is used as a carbon source.

The application discloses a fermentation L-histidine production strain and application thereof, and belongs to the technical field of bioengineering.The application takes E.coli CICC 10243 as a starting strain, integrates histidine synthesis operon genes hisDCB-cg2302-cg2301-HA-impA-FI, hisN and hisEG from corynebacterium glutamicum on the genome, introduces a pyruvate dehydrogenase AceE mutant coding gene, and inactivates the E.coli transcription factor SlyA coding gene slyA.The application prepares a new strain E.coli His08 for fermentation production of L-histidine, and the histidine yield reaches 58.39 g / L in a 5L fermenter for 60 h, and the sugar acid conversion rate is 0.15 g / g, so the application has important application value.

The invention relates to the field of biological pharmacy, in particular to product development and application of a targeted gender differential expression gene PDHB in fat reduction and obesity treatment. The invention provides a novel drug target, namely PDHB (pyruvate dehydrogenase E1 component beta subunit), which is closely related to obesity and sex difference, and is used for preparing or screening drugs capable of specifically reducing fat and treating obesity for visceral fat obesity (especially male) by improving mitochondrial energy metabolism.

Provided herein is a method of treating a patient having pyruvate dehydrogenase complex deficiency (PDCD), including administering to the patient an amount of triheptanoin effective to treat PDCD in the patient.