50 results about "Glycerol dehydratase" patented technology

Organisms are provided which express enzymes such as glycerol dehydratase, diol dehydratase, acyl-CoA transferase, acyl-CoA synthetase β-ketothiolase, acetoacetyl-CoA reductase, PHA synthase, glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase, which are useful for the production of PHAs. In some cases one or more of these genes are native to the host organism and the remainder are provided from transgenes. These organisms produce poly (3-hydroxyalkanoate) homopolymers or co-polymers incorporating 3-hydroxypropionate or 3-hydroxyvalerate monomers wherein the 3-hydroxypropionate and 3-hydroxyvalreate units are derived from the enzyme catalysed conversion of diols. Suitable diols that can be used include 1,2-propanediol, 1,3 propanediol and glycerol. Biochemical pathways for obtaining the glycerol from normal cellular metabolites are also described. The PHA polymers are readily recovered and industrially useful as polymers or as starting materials for a range of chemical intermediates including 1,3-propanediol, 3-hydroxypropionaldehyde, acrylics, malonic acid, esters and amines.

Recombinant organisms are provided comprising genes encoding aquacobalamin reductase, cob(II)alamin reductase, cob(I)alamin adenosyltransferase, glycerol dehydratase and 1,3-propanediol oxidoreductase activities useful for the production of 1,3-propanediol from a variety of carbon substrates. More specifically the following nucleotide sequences are provided: btuR, encoding the E. coli cob(I)alamin adenosyltransferase enzyme; cobA, encoding the Salmonella typhimurium cob(I)alamin adenosyltransferase enzyme; cobO, encoding the Pseudomonas denitrificans cob(I)alamin adenosyltransferase enzyme; dhaB1, encoding the α subunit of the Klebsiella glycerol dehydratase enzyme; dhaB2, encoding the β subunit of the Klebsiella glycerol dehydratase enzyme; dhaB3, encoding the γ subunit of the Klebsiella glycerol dehydratase enzyme; dhaT, encoding Klebsiella oxidoreductase enzyme; the yciK gene isolated from E. coli; the glucose isomerase promoter sequence from Streptomyces; and the N-terminal amino acid sequence for cob(II)alamin reductase from Pseudomonas denitrificans.

A method for producing 3-hydroxypropionaldehyde from glycerin in high conversion ratio is provided. The method is characterized by comprising a step of dehydrating glycerin using a microbial cell and/or a treated microbial cell containing diol dehydratase and/or glycerol dehydratase, and optionally diol dehydratase reactivating factor and/or glycerol dehydratase reactivating factor, under conditions so as to give a value (X/Y²) calculated by dividing a catalytic amount [X (U/g glycerin)] of diol dehydratase and/or glycerol dehydratase by square of glycerin concentration [Y (g/100 ml)] within a range of 10 to 8,000, to produce 3-hydroxypropionaldehyde.

The invention provides a method for producing 1,3-propylene glycol through fermentation via recombinant microbes. According to the invention, an endogenous dihydroxypropanone phosphatein phosphatase gene hdpA is over-expressed in Corynebacterium glutamicum to reinforce removal of phosphoric acid from dihydroxypropanone phosphatein so as to produce dihydroxypropanone; exogenous glycerol dehydrogenase is introduced to convert dihydroxypropanone into glycerin; and glycerin finally produces 1,3-propylene glycol under the action of exogenous glycerol dehydratase and an activator thereof and alcohol dehydrogenase. Corynebacterium glutamicum can use different cheap raw materials for fermentation, and cheap corn steep liquor can be used as a nutritional component to replace expensive yeast powder, so cost for raw materials is further reduced, and the problems in biosecurity and tolerance of the bacterial strain to a substrate and a product are overcome; and thalli obtained in the process of fermentation can be used as a product for a feed additive. The method provided by the invention produces few by-products and can further simplify the separating process of 1,3-propylene glycol.

The invention discloses recombinant Escherichia coli and application of the recombinant Escherichia coli in synthesizing 3-hydroxypropionic acid. The recombinant Escherichia coli is formed by glycerol dehydratase genes dhaB123, glycerol dehydratase reactivating factor genes gdrA, alpha-oxoglutarate semialdehyde dehydrogenase mutant coding genes TUkgsadh and glycerol-3-phosphate dehydrogenase coding genes gpdl which are guided into host bacteria. By means of the gene knockout technique, the yield of byproduct 1,3-propylene glycol is reduced, and meanwhile, cofactors NAD+ of aldehyde dehydrogenase are regenerated. Thus, the yield of the 3-hydroxypropionic acid is improved by 1.5 times.

The invention relates to new glycerin dehydrase gene cloning and its expression in relative host. The enzyme gene is cloned form clostridium perfringens which can transform the glycerin into 3-hydroxyl propionic aldehyde at normal biological chemical reacting condition. Based on the invention, it can recombine the 1,3-propylene glycol redox enzyme relative gene into Escherichia coli to gain a new metabolism engineering fungus which can be transformed into 1, 3-propylene glycol by fermenting tapioca hydrolyzate, glucose, or glycerol.

The invention discloses a method for co-production of 1,3-propanediol and glutamic acid through recombined corynebacterium glutamicum. In order to obtain the recombined corynebacterium glutamicum, the method comprises the following steps that 1, the encoding gene of 1,3-propanediol dehydrogenase is introduced into the corynebacterium glutamicum; 2, 1,2-propanediol dehydratase and the encoding gene of an activity factor of the 1,2-propanediol dehydratase are introduced into the corynebacterium glutamicum, or glycerol dehydratase and the encoding gene of an activity factor of the glycerol dehydratase are introduced into the corynebacterium glutamicum. By the adoption of the method, the corynebacterium glutamicum is used as a production strain, so that the problem of biological safety is solved, and the thallus processing cost is lowered. According to the method, in the production process that the glutamic acid and the 1,3-propanediol are fully coupled, reducing power can be fully utilized. The yield of the glutamic acid is not lowered, and meanwhile extremely-high mass conversion rate of the 1,3-propanediol to glycerinum is obtained. Moreover, the number of reaction by-products is small, and the separating process is simple and convenient.

The invention provides a glycerol dehydratase gene for coding glycerol dehydratase, a recombinant vector containing the gene, a recombinant gene engineering bacteria obtained by the transformation ofthe recombinant vector and an application thereof for preparing the recombinant glycerol dehydratase. The glycerol dehydratase gene can obtain an intracellular expression recombinant plasmid or a secretory expression recombinant plasmid by connection and construction with an expression vector, then the intracellular expression recombinant plasmid or the secretory expression recombinant plasmid isrespectively and correspondingly transformed to an escherichia coli strain and obtains the recombinant escherichia coli which contains recombinant glycerol dehydratase, and can recombine the escherichia coli to carry out biocatalysis and transformation for an enzyme source. As an enzyme for transformation, the recombinant glycerol dehydratase can respectively adopt glycerol, 1, 2-propylene glycolor alcohol as primer and carry out transformation reaction so as to prepare corresponding 3-hydroxypropionaldehyde, propionaldehyde or aldehyde and the like.

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.

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 discloses a genetically engineered bacterium used for producing 3-hydroxypropionic acid (3-HP) by fermentation, a constructing method and application of the genetically engineered bacterium. An acetaldehyde dehydrogenase gene in Saccharomyces cerevisiae and a glycerol dehydratase gene in Klebsiella pneumoniae are connected onto a prokaryotic expression vector pET30a(+) so as to construct a Pet30a-ALDH-DHAB double-gene prokaryotic expression vector, and then the constructed Pet30a-ALDH-DHAB double-gene prokaryotic expression vector is transformed in to an escherichia coli BL21(DE3) competent cell to construct the genetically engineered bacterium used for effectively producing the 3-HP by fermentation. The problems of high energy consumption and sever pollution in chemical industrial synthesis of the 3-HP and the problem of low yield rate in microbial fermentation production of the 3-HP are solved. By utilizing the genetically engineered bacterium to produce the 3-HP by fermentation, a yield of the 3-HP can reach to 8.443g/L, and a production process is simple and convenient for industrialized application.

The invention relates to a gene regulation continuous fermentation hybrid power system. The system solves a glycerin microorganism fermentation control problem by utilizing an optimal scientific theory. The glycerin microorganism fermentation control problem comprises control factors; and the control factors comprise biomass, extracellular glycerin, extracellular 1,3-PD, acetic acid, ethanol, intracellular glycerin, intracellular 3-HPA, intracellular 1,3-PD, mR, free regulatory proteins R, MGDHt, glycerol dehydratase GDHt, MPDOR and 1,3-PD oxidordeuctase PDOR. According to the gene regulation continuous fermentation hybrid power system, balance is found between mathematical modeling and numerical value accuracy, a possible metabolism path is proposed, a 14-dimension nonlinear hybrid power system with discrete path parameters and continuous system parameters for continuous fermentation is established, and an optimal control problem is structured by using a constraint condition.

The invention relates to the clone of a novel glycerol dehydratase activating factor gene and the expression thereof in related host. The activating factor gene, cloned from clostridium perfringens, can activate an inactive glycerol dehydratase under normal biochemical reaction conditions and recover the enzyme activity of the glycerol dehydratase, and can also enable the glycerol dehydratase to be difficult to be inactivated when the glycerol dehydratase is transformed to glycerin in the reaction. On the basis of the invention, related enzyme genes, such as the glycerol dehydratase, 1, 3-propanediol oxidoreductase, can be restructured to colon bacillus so as to get novel metabolic engineering bacteria. The metabolic engineering bacteria transform starch, glycerin or glucose to 1, 3-propanediol through fermentation.

The invention discloses a method for producing 1,3-propanediol by biologically catalyzing glycerol. According to the method, firstly, the glycerol is converted to an intermediate 3-hydroxypropionaldehyde through glycerol dehydratase, and under coupling catalysis of nicotinamide adenine dinucleotide (NAD) analog dependent 1,3-propanediol dehydrogenase and NAD analogue dependent oxidoreductase of regenerative NAD analogs, the intermediate produces the 1,3-propanediol. The catalytic system can be used for catalyzing the glycerol as a biodiesel by-product in an extracellular cell-free system or amicroorganism to produce the 1,3-propanediol, so that the production cost of the 1,3-propanediol is obviously reduced, and the product yield is increased.

The invention discloses a process for preparing 3-hydroxy-propionaldehyde, which adopts a two-stage process of cultivating microbial thalli firstly and then utilizing the thalli to transform glycerol, and the process comprises the following steps of: (1) preparing an MRS culture medium containing glucose or fructose for cultivating microorganisms that contain glycerol anhydrase and can secrete the 3-hydroxy-propionaldehyde to the outside of cells, and acquiring mass thalli; and (2) utilizing the thalli to serve as a biocatalyst for transforming the glycerol so as to produce the 3-hydroxy-propionaldehyde. The process has advantages of simple separation process, only needing the thalli separation in the transformation process, having no byproduct, saving separation and purification costs, reusable thalli after filtration, washing and activation, reducing the transformation time and improving the production efficiency.