169 results about "Dehydratase" patented technology

A non-naturally occurring microbial organism has cyclohexanone pathways that include at least one exogenous nucleic acid encoding a cyclohexanone pathway enzyme. A pathway includes a 2-ketocyclohexane-1-carboxyl-CoA hydrolase (acting on C—C bond), a 2-ketocyclohexane-1-carboxylate decarboxylase and an enzyme selected from a 2-ketocyclohexane-1-carboxyl-CoA hydrolase (acting on thioester), a 2-ketocyclohexane-1-carboxyl-CoA transferase, and a 2-ketocyclohexane-1-carboxyl-CoA synthetase. A pathway includes an enzyme selected from a 6-ketocyclohex-1-ene-1-carboxyl-CoA hydrolase (acting on C—C bond), a 6-ketocyclohex-1-ene-1-carboxyl-CoA synthetase, a 6-ketocyclohex-1-ene-1-carboxyl-CoA hydrolase (acting on thioester), a 6-ketocyclohex-1-ene-1-carboxyl-CoA transferase, a 6-ketocyclohex-1-ene-1-carboxyl-CoA reductase, a 6-ketocyclohex-1-ene-1-carboxylate decarboxylase, a 6-ketocyclohex-1-ene-1-carboxylate reductase, a 2-ketocyclohexane-1-carboxyl-CoA synthetase, a 2-ketocyclohexane-1-carboxyl-CoA transferase, a 2-ketocyclohexane-1-carboxyl-CoA hydrolase (acting on thioester), a 2-ketocyclohexane-1-carboxylate decarboxylase, and a cyclohexanone dehydrogenase. A pathway includes an adipate semialdehyde dehydratase, a cyclohexane-1,2-diol dehydrogenase, and a cyclohexane-1,2-diol dehydratase. A pathway includes a 3-oxopimelate decarboxylase, a 4-acetylbutyrate dehydratase, a 3-hydroxycyclohexanone dehydrogenase, a 2-cyclohexenone hydratase, a cyclohexanone dehydrogenase and an enzyme selected from a 3-oxopimeloyl-CoA synthetase, a 3-oxopimeloyl-CoA hydrolase (acting on thioester), and a 3-oxopimeloyl-coA transferase. Each these pathways can include a PEP carboxykinase. A method for producing cyclohexanone includes culturing these non-naturally occurring microbial organisms.

Methods for the fermentive production of four carbon alcohols are provided. Specifically, butanol, preferably 2-butanol is produced by the fermentive growth of a recombinant bacteria expressing a 2-butanol biosynthetic pathway. The recombinant microorganisms and methods of the invention can also be adapted to produce 2-butanone, an intermediate in the 2-butanol biosynthetic pathways disclosed herein. Specifically disclosed herein are the use of coenzyme B12-independent butanediol dehydratases that catalyzes the substrate to product conversion of 2,3-butanediol to 2-butanone in the process of producing 2-butanol and 2-butanone.

The invention discloses a method for extracting soybean grease from a soybean emulsion, which comprises the following steps of (1) carrying out the demulsification preprocessing of the soybean emulsion by utilizing ultrasonic processing parameters, centrifuging and collecting the supernatant; (2) adding ethanol to the supernatant for demulsification processing, centrifuging and collecting the supernatant to obtain the soybean grease. The method for assisting ethanol demulsification by utilizing ultrasonic can demulsificate the emulsion formed in the extraction course of a soybean dehydratase method so as to obtain high-quality soybean grease. The method has the advantages of simple required equipment, safe operation, no solvent residual of soybean oil, high-quality and high-nutritive-value grease obtainment. The recovery rate after demulsification under the demulsification technological conditions can reach about 98 to 100% through verification and comparison tests.

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.

Novel enzymes and novel enzymatic pathways for the pyruvate-based synthesis of shikimate or at least one intermediate thereto or derivative thereof, nucleic acids encoding the enzymes, cells transformed therewith, and kits containing said enzymes, cells, or nucleic acid. A KDPGal aldolase is used to perform condensation of pyruvate with D-erythrose 4-phosphate to form 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP); a 3-dehydroquinate synthase is used to convert the DAHP to 3-dehydroquinate (DHQ); DHQ dehydratase can then convert DHQ to the key shikimate intermediate, 3-dehydroshikimate.

Belonging to the technical field of industrial microbiology, the invention discloses a saccharomyces cerevisiae and application thereof in food fermentation. The saccharomyces cerevisiae strain 11-1 is preserved in China Center for Type Culture Collection on September 11, 2017 at Wuhan University, Wuhan, China, and the preservation number is CCTCC NO:M 2017488. Sense mutation occurs to the key gene pha2 in the path of synthesizing beta-phenylethyl alcohol from the strain, and the catalytic ability of mutated prephenate dehydratase PHA2P is strengthened by 22%. The invention also finds that thesaccharomyces cerevisiae pha2p suffers from the feedback inhibition of L-phenylalanine, when the strain is used for fermenting yellow rice wine, the alcoholic strength can reach 13 degrees, and the yield of beta-phenylethyl alcohol can reach 120mg / L.

Provided is a method of producing 2′-fucosyllactose including culturing in a medium supplemented with lactose a recombinant Corynebacterium glutamicum transformed to express α-1,2-fucosyltransferase, transformed to express GDP-D-mannose-4,6-dehydratase, transformed to express GDP-L-fucose synthase, and transformed to express lactose permease, wherein the recombinant Corynebacterium glutamicum has phosphomannomutase and GTP-mannose-1-phosphate guanylyltransferase.

The invention discloses a 2 '-fucosyllactose high-yield strain as well as a preparation method and application thereof. The preservation number of the strain is CGMCC No.19557. Escherichia coli E.coliBL21 (DE3) is used as a production host, and phosphomannose mutase (manB), mannose-1-phosphate phosphate guanylyltransferase (manC), GDP-mannose-4, 6-dehydratase (gmd), GDP-L-fucose synthase (flc) and alpha-1,2- fucosyltransferase (futC) are overexpressed so as to construct a synthesis path of the 2'-FL; substrate supply is improved by overexpressing sucrose transporter and lactose transporter onthe basis, the production capacity of 2 '-FL is further improved, efficient expression of 2'-FL in escherichia coli is realized, the 2 '-FL high-yield strain is improved.

The present invention provides an improved method for the biological production of 1,3-propanediol from a fermentable carbon source in a single microorganism. In one aspect of the present invention, an improved process for the conversion of glucose to 1,3-propanediol is achieved by the use of an E. coli transformed with the Klebsiella pneumoniae dha regulon genes dhaR, orfY, dhaT, orfX, orfW, dhaB1, dhaB2, dhaB3, and orfZ, all these genes arranged in the same genetic organization as found in wild type Klebsiella pneumoniae. In another aspect of the present invention, an improved process for the production of 1,3-propanediol from glucose using a recombinant E. coli containing genes encoding a G3PDH, a G3P phosphatase, a dehydratase, and a dehydratase reactivation factor compared to an identical process using a recombinant E. coli containing genes encoding a G3PDH, a G3P phosphatase, a dehydratase, a dehydratase reactivation factor and a 1,3-propanediol oxidoreductase (dhaT). The dramatically improved process relies on the presence in E. coli of a gene encoding a non-specific catalytic activity sufficient to convert 3-hydroxypropionaldehyde to 1,3-propanediol.

Provided is a method for producing L-threonine using a microorganism. In this method, the threonine dehydratase (tdc) gene present in the genomic DNA of the microorganism is partially inactivated by recombinant technology. TDC gene specificity involved in one of four threonine metabolic pathways for a microorganism with enhanced activity of a threonine operon-containing enzyme and phosphoenolpyruvate carboxylase (ppc) gene The ground is inactivated, thereby significantly increasing the production of L-threonine.

The invention relates to a biological improvement synthesis method of glutaric acid. According to the present invention, the method is achieved by expressing 2-hydroxyglutaric acid dehydrogenase, glutaryl-coenzyme A transferase, 2-hydroxyglutaryl coenzyme A dehydratase, trans-CoA reductase and thioesterase in a recombinant host Escherichia coli, the recombinant Escherichia coli expressing the genes is subjected to fermentation production, and the generation of the target product glutaric acid and the by-products such as glutaconic acid and 2-hydroxyglutaric acid is successfully detected in the fermentation broth; and the constructed biological synthesis approach provides the new method for the utilization of the renewable resources to synthesize the glutaric acid.

Sequences of B12-dependent dehydratases with improved reaction kinetics are presented. Use of these B12-dependent dehydratases reduce the rate of the enzyme's suicide inactivation in the presence of glycerol and 1,3-propanediol. The enzymes were created using error-prone PCR and oligonucleotide-directed mutagenesis to target the DhaB1 gene, which encodes the α-subunit of glycerol dehydratase. Mutants with improved reaction kinetics were rapidly identified using high throughput assays.

The invention provides a non-naturally occurring microbial organism having a muconate pathway having at least one exogenous nucleic acid encoding a muconate pathway enzyme expressed in a sufficient amount to produce muconate. The muconate pathway including an enzyme selected from the group consisting of a beta-ketothiolase, a beta-ketoadipyl-CoA hydrolase, a beta-ketoadipyl-CoA transferase, a beta-ketoadipyl-CoA ligase, a 2-fumarylacetate reductase, a 2-fumarylacetate dehydrogenase, a trans-3-hydroxy-4-hexendioate dehydratase, a 2-fumarylacetate aminotransferase, a 2-fumarylacetate aminating oxidoreductase, a trans-3-amino-4-hexenoate deaminase, a beta-ketoadipate enol-lactone hydrolase, a muconolactone isomerase, a muconate cycloisomerase, a beta-ketoadipyl-CoA dehydrogenase, a 3-hydroxyadipyl-CoA dehydratase, a 2,3-dehydroadipyl-CoA transferase, a 2,3-dehydroadipyl-CoA hydrolase, a 2,3-dehydroadipyl-CoA ligase, a muconate reductase, a 2-maleylacetate reductase, a 2-maleylacetate dehydrogenase, a cis-3-hydroxy-4-hexendioate dehydratase, a 2-maleylacetate aminoatransferase, a 2-maleylacetate aminating oxidoreductase, a cis-3-amino-4-hexendioate deaminase, and a muconate cis / trans isomerase. Other muconate pathway enzymes also are provided. Additionally provided are methods of producing muconate.

The invention belongs to the field of genetic engineering and relates to a method for producing 2'-fucosyllactose, in particular to a method for synthesizing 2'-fucosyllactose by co-catalysis of a genetically engineered strain and enzyme. The method of the invention comprises the steps of using a recombinant strain of lactococcus lactis which efficiently expresses hexokinase, phosphomannomutase and mannose-1-phosphate guanyl transferase as a production strain, and using mannose as a substrate to synthesize GDP-mannose, and then using GDP-mannose 4,6-dehydratase, GDP-4-keto-6-deoxymannose 3,5-mutarotase / 4-reductase, alpha1,2-fucosyltransferase to catalyze GDP-mannose in vitro to synthesize the 2'-fucosyllactose, thus providing a new method for the industrial production of the 2'-fucosyllactose.