102 results about "Pyruvate decarboxylase" patented technology

Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2. The organisms may be utilized for example in thermophilic SSF and SSCF reactions performed at temperatures that are optimal for cellulase activity to produce near theoretical ethanol yields without expressing pyruvate decarboxylase.

The invention provides a genetically modified Cyanobacteria having a construct comprising DNA fragments encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adh) enzymes obtained from the Zymomonas mobilis plasmid pLOI295. The Cyanobacteria are capable of producing ethanol in recoverable quantities of at least 1.7 mumol ethanol per mg of chlorophyll per hour.

The subject invention concerns the transformation of Gram-positive bacteria with heterologous genes which confer upon these microbes the ability to produce ethanol as a fermentation product. Specifically exemplified is the transformation of bacteria with genes, obtainable from Zymomonas mobilis, which encode pyruvate decarboxylase and alcohol dehydrogenase.

The invention provides isolated nucleic acids molecules which encode pyruvate decarboxylase enzymes having improved decarboxylase activity, substrate affinity, thermostability, and activity at different pH. The nucleic acids of the invention also have a codon usage which allows for high expression in a variety of host cells. Accordingly, the invention provides recombinant expression vectors containing such nucleic acid molecules, recombinant host cells comprising the expression vectors, host cells further comprising other ethanologenic enzymes, and methods for producing useful substances, e.g., acetaldehyde and ethanol, using such host cells.

The invention discloses a creatine jubase detection reagent. The reagent disclosed by the invention consists of a reagent R1 and a reagent R2 according to a volume ratio being 4 to 1, wherein the reagent R1 consists of an imidazole buffer solution, glucose, nano particles, N-acetylcysteine, ethylenediaminetetraacetic acid disodium salt, adenosine diphosphate, coenzyme I, adenine ribonucleotide, pyruvate decarboxylase, 6-phosphogluconic dehydrogenase, hexokinase and sodium dodecyl benzene sulphonate; and the reagent R2 consists of an imidazole buffer solution, phosphocreatine, a preservative and sodium dodecyl benzene sulphonate. Pyruvate decarboxylase and gamma-Fe2O3 nano particles are added into the creatine jubase detection reagent disclosed by the invention, so that interfering substances in a serum sample can be effectively eliminated, metal ions in the serum sample are chelated by matching sodium dodecyl benzene sulphonate and ethylenediaminetetraacetic acid disodium salt, the influence on reaction enzymes is alleviated, and the reagent is a stable, accurate and sensitive detection reagent with high interference resistance.

The invention discloses recombinant escherichia coli for producing tyrosol and a construction method and application thereof and belongs to the technical field of bioengineering. The escherichia coliheterologously expresses a saccharomyces cerevisiae pyruvate decarboxylase gene ARO10* after codon optimization. According to the recombinant escherichia coli, the ARO10* gene is integrated while a lacI locus, a trpE locus, a pabB locus, a pabA locus and a pykF locus of an escherichia coli genome are deleted, and a strain containing multiple copies of the ARO10* gene is obtained. On the basis of the recombinant strain, the ARO10* gene is randomly integrated at the multiple loci, and the ARO10* gene is inserted at a yccX locus so that the strain with high tyrosol yield can be obtained. No inducer nor antibiotic is required for fermentation by using this strain. After 48 hours of fermentation, the yield of tyrosol can reach 28 mM.

The invention provides isolated nucleic acids molecules which encode pyruvate decarboxylase enzymes having improved decarboxylase activity, substrate affinity, thermostability, and activity at different pH. The nucleic acids of the invention also have a codon usage which allows for high expression in a variety of host cells. Accordingly, the invention provides recombinant expression vectors containing such nucleic acid molecules, recombinant host cells comprising the expression vectors, host cells further comprising other ethanologenic enzymes, and methods for producing useful substances, e.g., acetaldehyde and ethanol, using such host cells.

The present invention provides recombinant mircoorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and / or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2. Both organisms produce near theoretical ethanol yields without expressing pyruvate decarboxylase

The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and / or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

The invention discloses a creatine kinase isoenzyme detection reagent. The reagent consists of a reagent R1 and a reagent R2 according to the volume ratio of 4 to 1, wherein the reagent R1 consists of an imidazole buffer liquid, glucose, nano particles, N-acetylcysteine, sodium ethylene diamine tetracetate, adenosine diphosphate, cozymase I, ribonucleotide, pyruvate decarboxylase, glucose 6-phosphate dehydrogenase, hexokinase, a goat anti-human CK-M polyclonal antibody and trehalose; the reagent R2 consists of an imidazole buffer liquid, phosphocreatine, sodium dodecyl benzene sulfonate and a preservative. The reagent disclosed by the invention is a creatine kinase isoenzyme detection reagent which is stable, accurate, and high in anti-interference, and has very high clinical application value.

The present invention relates to a method of identifying a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in (the cytosol of) a yeast cell comprising: a) providing a mutated yeast cell comprising a deletion of at least one gene of the (PDH) by-pass, selected from the genes encoding the enzymes pyruvate decarboxylase (PDC), acetaldehyde dehydrogenase (ALD), and acetyl-CoA synthetase (ACS); b) transforming said mutated yeast cell with an expression vector comprising a heterologous nucleotide sequence encoding a candidate polypeptide having potential enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA; c) testing said recombinant mutated yeast cell for its ability to grow on minimal medium containing glucose as sole carbon source, and d) identifying said candidate polypeptide as a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in (the cytosol of) said yeast cell when growth of said cell is observed. The invention further relates to a method of producing a fermentation production such as butanol.

A metabolically enhanced cyanobacterial cell for the production of ethanol is provided. The metabolically enhanced cyanobacterial cell for the production of ethanol comprises at least one recombinant gene encoding a pyruvate decarboxylase enzyme (Pdc) converting pyruvate to acetaldehyde, and at least one recombinant gene encoding a first Zn2+ dependent alcohol dehydrogenase enzyme (Adh) converting acetaldehyde to ethanol. The invention also provides a method for producing the metabolically enhanced cyanobacterium, a method for producing ethanol with the metabolically enhanced cyanobacterium, and a method for screening of alcohol dehydrogenase enzyme expressing cyanobacterial strains for the presence of NADPH-dependent native alcohol dehydrogenase enzymes.