35 results about "Acetoacetyl-CoA" patented technology

This invention relates to a recombinant microorganism having improved butanol production capacity and butanol production with the use of such recombinant microorganism with good efficiency. In this invention, the acetoacetyl-CoA synthase gene encoding an enzyme capable of synthesizing acetoacetyl-CoA from malonyl-CoA and acetyl-CoA and a group of genes involved in butanol biosynthesis that enables synthesis of butanol from acetoacetyl-CoA are introduced into a host microorganism.

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 describes a process for preparing acetone starting from acetyl-coenzyme A comprising process steps A. enzymatic conversion of acetyl-CoA into acetoacetyl-CoA B. enzymatic conversion of acetoacetyl-CoA into acetoacetate and CoA and C. decarboxylation of acetoacetate to acetone and CO2, which is characterized in that the coenzyme A is not transferred in process step B to an acceptor molecule. In addition, process step B is surprisingly catalysed by enzymes of the classes of acyl-CoA thioesterase, acyl-CoA synthetase or acyl-CoA thiokinase.A completely novel metabolic pathway is concerned, because the enzymatic hydrolysis of acetoacetyl-CoA without simultaneous transfer of CoA to a receptor molecule has never previously been described for any microbial enzyme.

This invention is intended to produce 2-butanol with excellent productivity via a fermentation process. Recombinant microorganisms into which the acetoacetyl-CoA synthase gene and a group of genes (i.e., genes involved in 2-propanol synthesis) encoding a set of enzymes synthesizing 2-propanol from acetoacetyl-CoA have been introduced are cultured, so that, in addition to 2-propanol, 2-butanol is produced at a high level in a medium.

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.

This invention relates to a recombinant microorganism capable of producing isoprene and isoprene production with the use of such recombinant microorganism with good efficiency. In this invention, the acetoacetyl-CoA synthase gene encoding an enzyme capable of synthesizing acetoacetyl-CoA from malonyl-CoA and acetyl-CoA and one or more genes involved in isoprene biosynthesis that enables synthesis of isoprene from acetoacetyl-CoA are introduced into a host microorganism.

The invention provides a method for constructing a gene engineering strain to enhance the resistance of a 1, 3-propanediol (PDO) production strain, which belongs to the technical field of biochemical engineering and comprises the step of: introducing Beta-ketothiolase PhbA, NADPH-dependent acetoacetyl CoA, PhbB and polyhydroxyalkanoates synthases, PhbC gene to a wild bacteria which can produce PDO to accumulate high concentration of PHB while the bacterial strain produces PDO so as obviously improve the tolerance of the bacterial strain to glycerol and 3-hydroxypropionaldehyde; the method can also be used to co-produce PHB and PDO. The method has the advantages that the constructed gene engineering strain improves the resistance of the bacterial strain to the high concentration of glycedrol and the intermediate product 3-hydroxypropionaldehyde and reduces the abnormal fermentation in industrial production; in addition, high concentration of PHB can be accumulated while PDO is produced, and PHB can be used as a separated product, thereby promoting the added valve of products and the utilization rate of materials and reducing the production cost.

The invention discloses a method for improving ratio of target product (butanol) in the fermentation products of clostridium acetobutylicum. The method inhibits the generation of acetone by effectivelThe invention discloses a method for improving ratio of target product (butanol) in the fermentation products of clostridium acetobutylicum. The method inhibits the generation of acetone by effectively inhibiting the metabolic pathway from acetoacetyl-CoA to acetone in the clostridium acetobutylicum. The synthesis of acetone can be blocked by knocking out an acetoacetate decarboxylase gene so as ty inhibiting the metabolic pathway from acetoacetyl-CoA to acetone in the clostridium acetobutylicum. The synthesis of acetone can be blocked by knocking out an acetoacetate decarboxylase gene so as to cause the acetoacetate decarboxylase to be inactive, thereby effectively improving the ratio of butanol in the fermentation products.o cause the acetoacetate decarboxylase to be inactive, thereby effectively improving the ratio of butanol in the fermentation products.

A transformant capable of producing isopropanol which is constructed by transferring the following genes (a) to (d) into an aerobic bacterium or a facultative anaerobic bacterium:(a) a foreign gene which encodes an enzyme having acetyl-CoA acetyltransferase activity;(b) a foreign gene which encodes an enzyme having acetoacetyl CoA:acetate CoA-transferase activity;(c) a foreign gene which encodes an enzyme having acetoacetate decarboxylase activity; and(d) a foreign gene which encodes an enzyme having isopropanol dehydrogenase activity.

The invention discloses a technique for biotransforming CO2 into isopropyl alcohol by utilizing blue algae, and provides the following six methods for preparing recombined blue algae: (1) leading a CoA transferase gene, an acetoacetate decarboxylase gene and an alcohol dehydrogenase gene into the blue algae; (2) leading the CoA transferase gene and the acetoacetate decarboxylase gene into the blue algae; (3) leading a coded gene of the alcohol dehydrogenase into the recombined blue algae in (2); (4) leading an acetyl-CoA acetyltransferase gene, an acetoacetyl-CoA transferase gene, the acetoacetate decarboxylase gene and the alcohol dehydrogenase gene into the blue algae; (5) leading the acetyl CoA acetyltransferase gene, the acetoacetyl-CoA transferase gene and the acetoacetate decarboxylase gene into the blue algae; and (6) leading the alcohol dehydrogenase gene into the recombined algae generating acetone in (5). The technique provided by the invention is one of ideal paths for realizing the continuous and healthy development of renewable and clean energy by utilizing the recombined algae to produce acetone and the isopropyl alcohol.

A taxaceae 3-hydroxy-3-methyl-diacid coenzyme A synthetic protein coding sequence is disclosed. Separated DNA molecule includes: coding polypeptide ribonucleotide sequence with taxaceae tm-Hmgs protein activity, having at least 70% homology for the ribonucleotide sequence and SEQ ID NO.3 of ribonucleotide sequence from No.98-1528 of ribonucleotide, the ribonucleotide sequence hybridizing with SEQ ID NO.3 of ribonucleotide sequence from No.98-1528 of ribonucleotide under condition of 40-55oC. It can be used to improve content of taxol or its precursor and protect human health.

The expression vector includes: the nucleic acids coding for the polypeptides forming a polypeptide complex having an enzyme activity allowing acetoacetyl-CoA to be converted to acetoacetate; optionally, at least one nucleic acid coding for a polypeptide having an enzyme activity allowing acetoacetate to be converted to acetone; and at least one nucleic acid coding for a polypeptide having an enzyme activity allowing acetone to be converted to isopropanol; the expression of the nucleic acids being controlled by a single constitutive promoter located upstream of the abovementioned nucleic acids.

The invention describes a process for the preparation of acetone starting from acetyl-coenzyme A comprising the process steps A. enzymatic conversion of acetyl-CoA to give acetoacetyl-CoA B. enzymatic conversion of acetoacetyl-CoA into acetoacetate and CoA, and C. decarboxylation of acetoacetate to give acetone and CO2, which process is characterized in that, in process step B, the coenzyme A is not transferred to an acceptor molecule. Moreover, surprisingly, process step B is catalyzed by enzymes from the classes acyl-CoA thioesterase, acyl-CoA synthetase or acyl-CoA thiokinase. This is an entirely novel metabolic pathway since the enzymatic hydrolysis of acetoacetyl-CoA without the simultaneous transfer of CoA to a receptor molecule has never been described before for any microbial enzyme.

Disclosed is a method for producing a polyester copolymer comprising 3HB and LA with high efficiency by the microbial fermentation using a sugar as a raw material. Specifically disclosed is a method for producing a polyester copolymer comprising hydroxybutyric acid and lactic acid, which comprises the following steps 1) and 2): 1) culturing a genetically modified microorganism carrying a protein capable of catalyzing the reaction for transferring CoA into propionic acid and / or lactic acid, a protein capable of catalyzing the reaction for producing acetoacetyl-CoA from two acetyl-CoA molecules, a protein capable of catalyzing the reduction reaction of acetoacetyl-CoA, and a protein capable of catalyzing the synthetic reaction of a polyhydroxyalkanoic acid and comprising an amino acid sequence selected from the following amino acid sequences a) and b) in a culture medium containing a carbon source: a) an amino acid sequence having the substitution of at least one of the amino acid residues located at position-130, position-325, position-477 and position-481 by another amino acid residue in the amino acid sequence depicted in SEQ ID NO:2; and b) an amino acid sequence additionally having the deletion or substitution of at least one amino acid residue other than the amino acid residues located at position-130, position-325, position-477 and position-481 or the insertion of at least one amino acid residue in the protein defined in item a); and 2) collecting the polyester copolymer from the culture obtained in step 1). The method enables the efficient production of polyester copolymer composed of 3HB and LA by using an inexpensive carbon source as a raw material, and can achieve the reduction in the cost for production of a biodegradable plastic.

The object of the present invention is to provide a method for efficiently producing a polyester copolymer consisting of 3HB and LA via microbial fermentation with the use of a sugar as a starting material.The following is provided: a method for producing a polyester copolymer consisting of 3-hydroxybutyrate and lactate, which comprises the steps of:(1) culturing a recombinant microorganism having a protein capable of catalyzing a reaction of transferring CoA to propionic acid and / or lactate, a protein capable of catalyzing a reaction of forming acetoacetyl-CoA from two acetyl-CoA molecules, a protein capable of catalyzing a reaction of acetoacetyl-CoA reduction, and a protein capable of catalyzing a reaction of polyhydroxyalkanoate synthesis consisting of the following amino acid sequence (a) or (b) in a medium containing a carbon source:(a) an amino acid sequence derived from the amino acid sequence shown in SEQ ID NO: 2 by substitution of at least one of the amino acids at positions 130, 325, 477, and 481 with different amino acid(s); or(b) an amino acid sequence derived from the protein specified in (a) by additional deletion or substitution of at least one amino acid other than the amino acids at positions 130, 325, 477, and 481 or by insertion of at least one amino acid residue; and(2) collecting the polyester copolymer from the culture product obtained in step (1). According to the production method of the present invention, a polyester copolymer consisting of 3HB and LA can be efficiently produced using an inexpensive carbon source as a starting material, and thus the production cost of a biodegradable plastic can be reduced.

A transformant capable of producing isopropanol which is constructed by transferring the following genes (a) to (d) into a coryneform bacterium:(a) an exogenous gene which encodes an enzyme having acetyl-CoA acetyltransferase activity;(b) an exogenous gene which encodes an enzyme having acetoacetyl CoA: acetate CoA-transferase activity;(c) an exogenous gene which encodes an enzyme having acetoacetate decarboxylase activity; and(d) an exogenous gene which encodes an enzyme having isopropanol dehydrogenase activity.

A method of manufacturing a butanediol includes a step of culturing a microbe that contains a gene that codes acetoacetyl-CoA synthase that catalyzes a reaction that irreversibly produces acetoacetyl-CoA from acetyl-CoA and malonyl-CoA, acetoacetyl-CoA reductase that catalyzes a reaction that produces 3-hydroxybutyryl-CoA from acetoacetyl-CoA, and an enzyme that catalyzes a reaction for producing a butanediol from 3-hydroxybutyryl-CoA.

The invention provides microorganisms and methods for the production of polyhydroxybutyrate (PHB) from gaseous substrates. In particular, the invention provides a non-naturally occurring Wood-Ljungdahl microorganism comprising (a) an enzyme that converts acetyl-CoA to acetoacetyl-CoA, (b) an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and (c) an enzyme that converts 3-hydroxybutyryl-CoA to polyhydroxybutyrate, and methods related thereto.

The invention relates to 3-hydroxyl-3-mathylglutaryl-CoA reductase albumen encoding sequence of barbadosnut, which belongs to the technical field of molecular biology and gene engineering. The DNA molecule separation comprises the following steps: the nucleotide sequence of polypeptide with barbadosnutJcHMGRs albumen activity is encoded, and the nucleotide sequence has a homology of at least 70 percent with the nucleotide sequence with nucleotide ranging from bit 81-1832 in SEQ ID NO.1; the nucleotide sequence can be hybridized with the nucleotide sequence with nucleotide ranging from bit 81-1832 in SEQ ID NO.2 at 40-55 DEG C. The invention is an enzyme, namely, 3-hydroxyl-3-mathylglutaryl-CoA generated by catalyzing acetyl-CoA and acetoacetyl coenzyme A. The enzyme is favorable for reducing the content of phorbol ester or the prosoma thereof in barbadosnut and greatly helpful for cultivating quality variety of aseptic barbadosnut.