65 results about "Thiolase" patented technology

A non-naturally occurring microbial organism has at least one exogenous nucleic acid encoding a MEK pathway enzyme expressed in a sufficient amount to produce MEK. The MEK pathway includes an enzyme selected from an acetoacetyl-CoA dehydrogenase (bifunctional), an acetoacetyl-CoA aldehyde dehydrogenase, a 3-oxobutyraldehyde reductase, a 3-oxobutanol dehydratase, an MEK oxidoreductase, a 3-oxobutyraldehyde aminotransferase, a 4-aminobutan-2-one deaminase, a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP aminotransferase, a 2,4-dioxopentanoate decarboxylase, an AKP deaminase, an acetylacrylate decarboxylase, an AKP decarboxylase, a glutamate dehydrogenase, a 3-oxobutyraldehyde oxidoreductase (aminating) and an AKP oxidoreductase (aminating). A 2-butanol pathway further includes an MEK reductase. A method for producing MEK or 2-butanol includes culturing these organisms under conditions and for a sufficient period of time to produce MEK or 2-butanol.

The invention discloses recombinant saccharomycetes for high yield of sandalwood oil as well as a construction method and application of the recombinant saccharomycetes. The method comprises the following steps: 1) integrating genes such as acetoacetyl-CoA thiolase gene and the like which are derived from saccharomyces cerevisiae and are driven by a strong promoter and CYP450 reductase gene whichis driven by a weak promoter into a saccharomyces cerevisiae BY4742 genome of an original strain in a homologous recombination manner; 2) replacing a promoter of a squalene synthase gene in the recombinant bacteria; knocking out genes such as galactose modulin 80 genes; and (3) inserting a sandalene synthase gene driven by a strong promoter and a CYP450 monooxygenase gene into the multi-copy freeplasmid to obtain an expression plasmid, and introducing the expression plasmid into the recombinant saccharomyces cerevisiae obtained in the step (2) to obtain the recombinant saccharomyces cerevisiae for high yield of sandalwood oil. According to the method, the defect that sandalwood oil is extracted from sandalwood is overcome, and the yield of each liter of sandalwood oil of the recombinant yeast strain reaches 1g. Industrial production requirements can be met.

This document describes biochemical pathways for producing 1,3-butanediol using a polypetide having Beta-ketothiolase activity to form a 3-oxo-5-hydroxypentanoyl-CoA intermediate that can be enzymatically converted to 1,3-butanediol, as well as recombinant hosts producing 1,3-butanediol.

The invention provides a research and an application of beta-ketothiolase in extreme halophilic archaeon, wherein beta-ketothiolase is related to 3-HV monomer synthesis of a biodegradable material PHBV. The protein provided by the invention is a protein showed by the following description (1) or (2): (1) a protein composed of an amino acid sequence represented by a sequence 1 in the sequence table; (2) a protein formed after the amino acid sequence represented by the sequence 1 is processed after substitutions and / or deletions and / or additions of one or more amino acid residues, wherein the obtained protein is derived from the protein represented by (1), and is PHBV and / or 3-HV related. As a result of experiments, when the coding gene of the poly(3-hydrobutyrate-co-3-hydroxyvalerate) and / or 3-hydroxyalkanoate synthesis related protein is introduced into host bacteria, the molar ratio of generated 3-HV monomer for synthesizing PHBV is greater than 3-HV generated from the host bacteria.

This disclosure generally relates to the use of microorganisms to make various functionalized polyketides through polyketoacyl-CoA thiolase-catalyzed non-decarboxylative condensation reactions instead of decarboxylative reactions catalyzed by polyketide synthases. Native or engineered polyketoacyl-CoA thiolases catalyze the non-decarboxylative Claisen condensation in an iterative manner (i.e. multiple rounds) between two either unsubstituted or functionalized ketoacyl-CoAs (and polyketoacyl-CoAs) serving as the primers and acyl-CoAs serving as the extender unit to generate (and elongate) polyketoacyl-CoAs. Before the next round of polyketoacyl-CoA thiolase reaction, the β-keto group of the polyketide chain of polyketoacyl-CoA can be reduced and modified step-wise by 3-OH-polyketoacyl-CoA dehydrogenase or polyketoenoyl-CoA hydratase or polyketoenoyl-CoA reductase. Dehydrogenase converts the β-keto group to β-hydroxy group. Hydratase converts the β-hydroxy group to α-β-double-bond. Reductase converts the α-β-double-bond to single bond. Spontaneous or thioesterase catalyzed termination reaction terminates the elongation of polyketide chain of polyketoacyl-CoA at any point through CoA removal and spontaneous reactions rearrange the structure, generating the final functional polyketide products.

The invention discloses a method for preparing hydroxyalkanoate homopolymer and special bacteria thereof. Recombinant pseudomonas putida is prepared by the method comprising the following steps of: depriving coding functions of 3-hydroxyfatty acyl-coenzyme A dehydrogenase coding gene and 3-ketoacyl coenzyme A thiolase coding gene in pseudomonas putida to obtain recombinant pseudomonas putida, andrecording the recombinant pseudomonas putida as recombinant pseudomonas putida I. Experiments prove that the hydroxyalkanoate homopolymer, particularly medium and long-chain hydroxyalkanoate homopolymer (C4-C14), can be obtained by using the recombinant bacteria organism to synthesize polyhydroxyalkanoate. The obtained hydroxyalkanoate homopolymer has high purity and high yield. The method of theinvention can synthesize the medium and long-chain hydroxyalkanoate homopolymer with random length, and overcomes the defect that only one 3-hydroxybutyrate (HB) homopolymer can be obtained by biosynthesis in the prior art. Therefore, the method of the invention has broad application prospect in the biosynthesis field of the hydroxyalkanoate homopolymer.

An isopropyl alcohol-producing Escherichia coli equipped with an isopropyl alcohol production system, having at least one enhanced enzyme activity selected from the group consisting of an enhanced malate dehydrogenase activity, an enhanced NAD(P)+ transhydrogenase (AB-specific) activity, and an enhanced thiolase activity, and an isopropyl alcohol producing method including producing isopropyl alcohol from a plant-derived raw material using the isopropyl alcohol-producing Escherichia coli.

The invention discloses a method for producing adipic acid by utilizing saccharomyces cerevisiae, and belongs to the field of bioengineering. According to the invention, a saccharomyces cerevisiae BY4741 wild type strain or a saccharomyces cerevisiae BY4741 strain with LSC1 gene knocked out is used as a host; the beta-ketothiolase gene, the 3-hydroxyacyl-coenzyme A dehydrogenase gene, the 3-hydroxyadipoyl dehydrogenase gene, the 5-carboxyl-2-pentenyl coenzyme A reductase gene and the adipoyl coenzyme A in a T.fusca adipic acid reverse degradation pathway are overexpressed by modules, wherein the yield of adipic acid can reach 0.033-0.113 g / L. Due to the excellent stress resistance, acid resistance, hereditary stability and food safety of the saccharomyces cerevisiae, the invention providesa basis for industrial fermentation production of adipic acid by saccharomyces cerevisiae hosts.

The invention discloses a poly-3-hydroxy propionic acid copolymer and a production method thereof and belongs to the technical field of genetic engineering. According to the poly-3-hydroxy propionic acid copolymer and the production method thereof, a glycerol dehydratase gene and a glycerol dehydratase re-activating enzyme gene are integrated with a host strain genome by a gene integration technology, a polyhydroxy fatty acid synthase gene, a propionaldehyde dehydrogenase gene, a beta-ketoacyl coenzyme A thiolase gene, an acetoacetyl coenzyme A reductase gene and a propionyl coenzyme A synthetase gene are introduced, and a recombinant gene engineering strain has the ability of biologically synthesizing poly-3-hydracrylic acid-co-3-hydroxyvaleric acid. According to the poly-3-hydroxy propionic acid copolymer and the production method thereof, the poly-3-hydracrylic acid-co-3-hydroxyvaleric acid is obtained in a biosynthesis manner for the first time; compared with poly-3-hydracrylic acid, the obtained poly 3-hydracrylic acid-co-3-hydroxyvaleric acid has a higher melting point and lower crystallinity, has good degradability and can serve as a packaging material, a medical implant material, a drug sustained-release material and an electrochemical material.

The present invention relates to a modified thiolase protein with an improved activity, a polynucleotide encoding the modified protein, an expression vector including the polynucleotide, and a transformant, to a composition for producing a biobutanol including the thiolase with an improved activity or a cell expressing the thiolase, and to a method of producing the biobutanol.

The invention discloses a difunctional thiolase gene MvaE. The invention further discloses difunctional thiolase, an expression cassette, a recombinant vector, recombinant bacteria or cells and application thereof. The invention further discloses a method for producing fatty acid. The difunctional thiolase gene MvaE disclosed by the invention can utilize rich culture media containing glucose, glycerol or other carbon sources as raw materials under facultative anaerobic conditions, so that the fatty acid content of C<14:0>, C<16:0> and C<18:0> in host bacteria can be remarkably increased, and the increasing amplitude of the difunctional thiolase gene MvaE is different from 27% to 213% according to different host strains.