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37 results about "Methanol dehydrogenase" patented technology

In enzymology, a methanol dehydrogenase is an enzyme that catalyzes the chemical reaction: methanol ⇌ formaldehyde + 2 electrons + 2H⁺ How the electrons are captured and transported depends upon the kind of methanol dehydrogenase and there are two main types. A common electron acceptor in biological systems is nicotinamide adenine dinucleotide (NAD⁺) and some enzymes use a related molecule called nicotinamide adenine dinucleotide phosphate (NADP⁺).

Multicopy-integration of heterologous genes and expression in methylobacterium

The integration of genes into methylobacterium, such as M. extorquens ATCC 55366 is disclosed, using a transposon system, preferably the mini-Tn7 transposon system, and under the control of a promoter, such as the strong methanol dehydrogenase promoter (PmxaF). Multicopy integration of genes of interest is also disclosed. The unique and specific attachment site for the Tn7 attachment (attTn7) is identified for M. extorquens.
Owner:NAT RES COUNCIL OF CANADA

Method for producing target substance

The present invention discloses a method for producing a target substance using a coryneform bacterium comprising culturing a coryneform bacterium having an ability to produce the target substance in a medium, resulting in accumulation of the target substance in the medium or cells of the bacterium, and collecting the target substance from the medium or the cells of the bacterium. Also disclosed is a coryneform bacterium which is introduced with a methanol dehydrogenase gene and which has enhanced activities of hexulose phosphate synthase and phosphohexuloisomerase, and to which an ability to utilize methanol is imparted or which has enhanced ability to utilize methanol, and the medium contains methanol as a carbon source.
Owner:AJINOMOTO CO INC

Recombinant bacterium with granular methane monooxygenase activity and application thereof

InactiveCN1329506CSolve heterologous expression problemsAvoid defectsBacteriaEnzymesBiotechnologyEpoxy
The present invention discloses one kind of recombinant bacterium with granular methane monooxygenase activity and its application. Fusion gene comprising the promoter sequences of coding gene and desulfurizing gene of granular methane monooxygenase is introduced into Rhodococcus, and after screening, the strain expressing granular methane monooxygenase or the recombinant bacterium with granular methane monooxygenase activity is obtained. The promoter sequence of the desulfurizing gene is located in the upstream of the granular methane monooxygenase coding gene. The recombinant bacterium avoids the effect of methanol dehydrogenase radically, makes methane oxidation stop in methanol reaction step, and is superior to wild methane oxidizing bacteria and easy to culture. The recombinant bacterium can convert alkane in natural gas into liquid alcohol, convert methane into methanol and oxidize propylene into 1, 2-epoxy propane.
Owner:TSINGHUA UNIV

Method for producing target substance

In a method for producing a target substance by using a microorganism comprising culturing a microorganism having an ability to produce the target substance in a medium to produce and accumulate the target substance in the medium or cells of the microorganism and collecting the target substance from the medium or the cells of the microorganism, there are used, as the microorganism, a microorganism to which a methanol dehydrogenase gene is introduced, of which activities of hexulose phosphate synthase and phosphohexuloisomerase are enhanced and which is modified so that an ability to utilize methanol should be imparted or enhanced, and there is used a medium containing methanol as a carbon source.
Owner:AJINOMOTO CO INC

Escherichia coli genetically engineered bacteria for preparing isoprene through catalytic methanol and preparation method and application of escherichia coli genetically engineered bacteria

The invention discloses escherichia coli genetically engineered bacteria for preparing isoprene through catalytic methanol and a preparation method and application of the escherichia coli genetically engineered bacteria, and belongs to the technical field of genetic engineering. The escherichia coli genetically engineered bacteria have the isoprene synthesizing approach, and meanwhile methanol dehydrogenase gene MDH, 3-hexulose-6-P synthase gene RmpA, 3-hexulose-6-P isomerase gene RmpB and methanol dehydrogenase activated protein gene Act are overexpressed. Meanwhile, the invention further provides a method and application method for establishing the escherichia coli genetically engineered bacteria. The escherichia coli genetically engineered bacteria can biologically synthesize isoprene with methanol as the substrate and has the advantages of being high in conversion efficiency, low in production cost and the like; the conversion rate of methanol-isoprene can reach 21.83%.
Owner:QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI +1

Method for producing methanol by methane bioconversion

The invention discloses a method for producing methanol by methane bioconversion. In the method, methylosinus trichosporium is used as biocatalyst, methanol containing gas and oxygen-containing gas are introduced into solution containing the methylosinus trichosporium and phosphate through a breathable film to react so as to prepare high-concentration methanol solution. The method realizes methanol production by methane bioconversion by using the high-concentration phosphate as methanol dehydrogenase inhibitor to inhibit further oxidation of the methane, realizes high-concentration methanol production by using high cell concentration methylosinus trichosporium as the biocatalyst, avoids unsafe hidden danger possibly generated by mixing of the methane and oxygen in a bubbleless film aeration mode, and reinforces the gas-liquid mass transfer process of the methane and the oxygen. By adopting the method, high concentration methanol solution can be safely and effectively produced, so the invention provides a reliable technology for industrialization of methanol production by methane bioconversion.
Owner:GRADUATE SCHOOL OF THE CHINESE ACAD OF SCI GSCAS +1

Construction method of oil-producing yeast engineering strain by using methyl alcohol and application

The invention discloses a construction method of an oil-producing yeast engineering strain by using methyl alcohol and application. The strain construction method disclosed by the invention relates tosimultaneous overexpression of methanol dehydrogenase, hexose phosphatase and oxoisomerase which are key enzymes used by the methyl alcohol in oil-producing yeast. An obtained gene engineering straincan efficiently make use of the methyl alcohol to produce grease, so as to provide a core technology for conversion of the methyl alcohol into long carbon chain fuel and a functional chemical. The construction method has a wide industrial application prospect.
Owner:DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI

Synthetic methylotrophy to liquid fuels and chemicals

A non-naturally occurring microbe capable of growing in a medium comprising methanol is provided. The methanol contributes to a significant percentage (e.g., at least 40%) of the carbon source for the non-naturally occurring microbe, which expresses heterologous methanol dehydrogenase (MDH) and heterologous ribulose monophosphate (RuMP) pathway enzymes. Methods for producing liquid fuels and chemicals by the non-naturally occurring microbe and methods for preparing the non-naturally occurring microbe are also provided.
Owner:UNIVERSITY OF DELAWARE

Method for reducing NAD analogue by using methanol

The invention discloses a method for reducing an NAD analogue by using methanol and application of the method. In the method, the methanol is taken as a reducing agent, methanol dehydrogenase capableof utilizing the methanol is taken as a catalyst, and the NAD analogue is transformed into a reduction state while the methanol is oxidized by the methanol dehydrogenase. The method can be used for producing a reduction-state NAD analogue or a deuterated reduction-state analogue, and can also provide reduction-state coenzyme for an enzymatic reaction consuming the reduction-state NAD analogue; thereduction-state NAD analogue can be used as a coenzyme to be applied to reduction reactions catalyzed by enzymes such as malic enzyme ME-L310R / Q401C, D-lactic dehydrogenase DLDH-V152R, and saccharomyces cerevisiae alcohol dehydrogenase, and wide application of the NAD analogue is facilitated. According to the NAD analogue reduction method, the reduction-state NAD analogue can be regenerated undera mild condition for preparing malic acid or lactic acid; and the reduction-state NAD analogue can also serve as an oxidation-reduction force to regulate the metabolic intensity of malic acid or lactic acid in microorganisms.
Owner:DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI

Method for improving activity of rate-limiting enzyme in methanol metabolism process of escherichia coli

The invention discloses a method for improving the activity of rate-limiting enzyme in a methanol metabolism process of escherichia coli. After genes Mdh and Hps-Phi are overexpressed by the escherichia coli, the enzyme activity of methanol dehydrogenase is improved by adding activator protein nudF, so that the metabolic rate of methyl alcohol is increased; the genes Mdh and Hps-Phi are from Mdh2 and Hps-Phi of bacillus methanol MGA3; the activator protein nudF is from E.coli MG1655. According to the method, a methanol metabolism path is assembled in the escherichia coli, so that methanol metabolism is realized; furthermore, the enzyme activity of the methanol dehydrogenase Mdh is improved through molecular improvement, thus realizing enhancement of the methanol metabolism.
Owner:NANJING UNIV OF TECH

Synthetic methylotrophy to liquid fuels and chemicals

A non-naturally occurring microbe capable of growing in a medium comprising methanol is provided. The methanol contributes to a significant percentage (e.g., at least 40%) of the carbon source for the non-naturally occurring microbe, which expresses heterologous methanol dehydrogenase (MDH) and heterologous ribulose monophosphate (RuMP) pathway enzymes. Methods for producing liquid fuels and chemicals by the non-naturally occurring microbe and methods for preparing the non-naturally occurring microbe are also provided.
Owner:UNIVERSITY OF DELAWARE

Method for producing 1,3-propanediol by using methanol/formaldehyde and glucose as common substrates

The present invention belongs to the technical field of microbial fermentation and particularly relates to a method for producing 1,3-propanediol by using methanol / formaldehyde and glucose as common substrates. Four enzymes are over-expressed in recombinant Escherichia coli: methanol dehydrogenase (MDH), aldolase, 2-oxo decarboxylase and 1,3-propanediol dehydrogenase. A novel methanol and formaldehyde C1 compound converted 1,3-PDO synthetic pathway is established. The method is used to solve problems that utilization of methanol, formaldehyde and other C1 compounds is too dependent on ribulosemonophosphate pathway (RuMP) and regeneration of Ru5P receptors is inefficient, avoids an addition of a cofactor vitamin B12 or S-adenosylmethionine (SAM), also shortens the 1,3-PDO synthetic pathwayand increases yield of 1,3-PDO.
Owner:HUAANTANG BIOTECH GRP CO LTD

Recombinant pseudomonas putida and application

The invention relates to recombinant pseudomonas putida and application thereof to production of 5-methyl pyrazine-2-carboxylic acid, and belongs to the field of fermenting engineering. The recombinant pseudomonas putida JN-18 is characterized in that the collection number is CGMCC No. 15734; the collection unit is Common Microorganism Center of China Committee for Culture Collection of Microorganisms. The recombinant pseudomonas putida has the advantages that the over-expression of xylene monooxygenase, benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase from self endogenous plasmid in P.putida ATCC 33015 is successfully realized; P.putida JN-18 is used for catalyzing 2,5-dimethylpyrazine to produce 5-methyl pyrazine-2-carboxylic acid in a whole-cell way.
Owner:DISHA PHARMA GRP +2

Recombinant escherichia coli for producing 5-methylpyrazine-2-carboxylic acid

The invention relates to recombinant escherichia coli for producing 5-methylpyrazine-2-carboxylic acid, in particular to a method for increasing 5-methylpyrazine-2-carboxylic acid by adjusting the proportion of dimethylbenzene monooxygenase double subunits. According to the adjustment of the proportion of the dimethylbenzene monooxygenase double subunits, escherichia coli (E.coli BL21 (DE3) is used as an original strain, a 5-methylpyrazine-2-carboxylic acid synthetic route is constructed by utilizing CRISPR / cas9 homologous recombination to integrate xylene monooxygenase, benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase genes. According to the strain disclosed by the invention, the yield of the 5-methylpyrazine-2-carboxylic acid in recombinant escherichia coli can be increased to15.6 g / L, and a foundation is laid for industrial production of the 5-methylpyrazine-2-carboxylic acid.
Owner:迪嘉药业集团股份有限公司

Cell-based systems for production of methyl formate

Disclosed is a process in which a recombinant organism, such as a yeast, expressing a heterologous S-adenosylmethionme (SAM)-dependent methyl halide transferase (MHT) protein is combined with a halide and a carbon source in a cultivation medium under conditions in which methyl formate is produced. The cell may genetically modified to express methyl formate synthase, methanol dehydrogenase and / or hydrolytic dehalogenase at levels higher than a cell of the same species that is not genetically modified. The methyl formate may be collected and used in a variety of applications. The halide may be chlorine, bromine or iodine.
Owner:RGT UNIV OF CALIFORNIA

Nucleic acids and vectors for use with methanotrophic bacteria

ActiveUS20170121718A1VectorsOxidoreductasesMethanotrophic bacteriumGenetics manipulation
The present disclosure provides nucleic acids and vectors for use with methanotrophic bacteria. Related host cells and methods for using such nucleic acids and vectors for expressing polypeptides or other genetic manipulation of methanotrophic bacteria are also provided. In one aspect, the present disclosure is directed to a non-naturally occurring nucleic acid molecule, comprising (1) a promoter that is functional in a methanotrophic bacterium, and (2) a native or altered methanol dehydrogenase (MDH) ribosomal binding sequence, provided that when the promoter is an MDH gene promoter, the nucleic acid comprises an altered MDH ribosomal binding sequence.
Owner:CALYSTA

Metal organic framework multi-enzyme composite catalytic material, preparation method, and application and use method of metal organic framework multi-enzyme composite catalytic material

The invention discloses a metal organic framework multi-enzyme composite catalytic material, a preparation method, and application and use method of the metal organic framework multi-enzyme composite catalytic material. The composite catalytic material comprises formate dehydrogenase, formaldehyde dehydrogenase, methanol dehydrogenase, UDP-glucose dehydrogenase and an MOF material, and is provided with a hollow capsule cavity; the formate dehydrogenase, the formaldehyde dehydrogenase and the methanol dehydrogenase are wrapped in the capsule cavity, and the UDP-glucose dehydrogenase is immobilized in the MOF material. According to the composite catalytic material of the invention, CO2 can be effectively recycled to provide a potential method for solving the greenhouse effect, repeated utilization can be implemented through immobilization and cyclic regeneration of cofactors while relatively high catalytic activity is kept, and a high-value industrial product is synthesized by a low-cost and environment-friendly method.
Owner:NANJING NORMAL UNIVERSITY

Synthetic methylotrophs

The present invention provides a non-naturally occurring microbe capable of growing in a medium comprising methanol, comprising a heterologous polynucleotide encoding a heterologous methanol dehydrogenase (MDH) derived from a Corynebacterium organism (Cor), wherein the MDH is expressed in the microbe, and wherein the MDH exhibits a Km of no more than 3 mM for methanol. Also provided are uses of the non-naturally occurring microbe for oxidizing methanol and producing a metabolite as well as the preparation of the non-naturally occurring microbe.
Owner:UNIVERSITY OF DELAWARE

Aldolase mutant and its application in production of 1, 3-propylene glycol

The invention relates to the technical field of synthetic biology and microbial fermentation, in particular to modification of deoxyribose-5-phosphate aldolase through semi-rational design and overexpression of four enzymes in recombinant escherichia coli, namely methanol dehydrogenase, ethanol dehydrogenase, deoxyribose-5-phosphate aldolase, 1, 3-propylene glycol oxidoreductase or 1, 3-propylene glycol oxidoreductase isozyme, to construct unnatural 1, 3-propylene glycol from methanol, a formaldehyde-carbon compound, ethanol and acetaldehyde. The activity of the DERATMAS233D mutant screened by the invention is improved by 51% compared with that of a wild type enzyme; When the conversion from formaldehyde and ethanol to PDO by using escherichia coli, the yield of PDO is increased by 21.8%; when the formaldehyde and the acetaldehyde are converted into the PDO, the yield of the PDO is increased by 9.2%, the synthesis route of the PDO is greatly shortened, and the yield of the PDO is increased.
Owner:BEIJING UNIV OF CHEM TECH

Methanol dehydrogenase fusion proteins

Described herein are fusion proteins including methanol dehydrogenase (MeDH) and at least one other polypeptide such as 3-hexulose-6-phosphate dehydrogenase (HPS) or 6-phospho-3-hexuloisomerase (PHI),such as DHAS synthase or fructose-6-Phosphate aldolase or such as DHA synthase or DHA kinase. In a localized manner, the fusion protein can promote the conversion of methanol to formaldehyde and thento a ketose phosphate such as hexulose 6-phosphate or then to DHA and G3P. When expressed in cells, the fusion proteins can promote methanol uptake and rapid conversion to the ketose phosphate or tothe DHA and D3P, which in turn can be used in a pathway for the production of a desired bioproduct. Beneficially, the rapid conversion to the ketose phosphate or to the DHA and G3P can avoid the undesirable accumulation of formaldehyde in the cell. Also described are engineered cells expressing the fusion protein, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.
Owner:GENOMATICA INC

Construction method and application of homoserine and threonine biosynthetic pathway

PendingCN114107408ABacteriaTransferasesSerine KinaseAcyl CoA dehydrogenase
The invention discloses construction of a new non-natural biosynthesis way of homoserine and threonine, and belongs to the technical field of industrial biology. The non-natural biosynthetic pathway comprises methanol dehydrogenase, aldolase, transaminase, homoserine kinase and threonine synthetase, and the pathway can be used for synthesizing homoserine or threonine by taking methanol and pyruvic acid as substrates. According to the present invention, the homoserine or the threonine can be synthesized by using methanol and pyruvic acid as the carbon source by constructing the in vitro multi-enzyme catalysis system and assembling in the microbial cell body through the pathway, and the pyruvic acid can be synthesized through glycolysis by using glucose as the substrate in the microbial cell body; therefore, homoserine or threonine is synthesized by taking methanol and glucose as carbon sources. According to the invention, the methanol-carbon compound with high energy density is introduced into a homoserine and threonine synthesis route, 200% of sugar-acid molar conversion rate can be theoretically realized, and the method has a great application prospect.
Owner:TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI

Methanol utilization

Described herein are enzymes, such as for example, methanol dehydrogenase (MDH), 3-hexulose-6-phosphate isomerase (PHI), 3-hexulose-6-phosphate synthase (HPS), ribose-5-phosphate isomerase (RPI), ribulose 5-phosphate 3-epimerase (RPE), transketolase (TKT), transaldolase (TAL) enzymes, phosphofructokinase (PFK), Sedoheptulose 1,7-Bisphosphatase (GLPX), fructose-bisphosphate aldolase (FBA), 6-phosphogluconate dehydrogenase (GND), and glucose-6-phosphate dehydrogenase (ZWF); recombinant host cells expressing the enzymes; methods of producing methylotrophic cells; and methods of producing amino acids (e.g., lysine).
Owner:GINKGO BIOWORKS INC

A method for producing 1,3-propanediol with methanol/formaldehyde and glucose as co-substrate

The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for producing 1,3-propanediol by using methanol / formaldehyde and glucose as co-substrates. The present invention constructs novel methanol and formaldehyde one-carbon by overexpressing four enzymes in recombinant Escherichia coli: methanol dehydrogenase (MDH), aldolase, 2-oxo decarboxylase and 1,3-propanediol dehydrogenase The compound is transformed into a 1,3‑PDO synthetic pathway. It is used to solve the problem that the utilization of one-carbon compounds such as methanol and formaldehyde is too dependent on the ribulose monophosphate pathway (RuMP) and the low-efficiency Ru5P receptor regeneration problem; avoiding the production pathway with additional cofactors vitamin B12 or S-adenosine A The problem of thionine (SAM), and shorten the synthesis pathway of 1,3-PDO, increase the yield of 1,3-PDO.
Owner:HUAANTANG BIOTECH GRP CO LTD

Recombinant microorganism capable of efficiently utilizing methanol and application thereof

The invention relates to the technical field of genetic engineering and biological fermentation, and particularly discloses a recombinant microorganism capable of efficiently utilizing methanol and application of the recombinant microorganism. The invention provides a recombinant microorganism which has increased expression and / or enzymatic activity of methanol dehydrogenase and dihydroxyacetone synthetase and reduced expression quantity of a formaldehyde dissimilatory gene frmAB and expression and / or enzymatic activity of 6-phosphofructokinase compared with an original strain, and the recombinant microorganism has the advantages that the expression quantity of the methanol dehydrogenase and the dihydroxyacetone synthetase is increased, and the expression quantity of the formaldehyde dissimilatory gene frmAB and the expression quantity of 6-phosphofructokinase are reduced; the starting strain is escherichia coli. Specifically, after a formaldehyde dissimilatory gene frmAB, a 6-phosphofructokinase gene pfkAB and an alpha-ketoglutarate dehydrogenase gene sucA are knocked out at the same time, and a 3-phosphoglyceraldehyde dehydrogenase gene gapA is inhibited, the utilization rate of methanol can be further improved, and accumulation of intermediate formaldehyde is reduced.
Owner:TSINGHUA UNIV
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