54 results about "Malic enzyme" patented technology

A nucleotide sequence encoding a malic enzyme and a method for preparing succinic acid using the same, more particularly, a maeB nucleotide sequence encoding a malic enzyme B having the activity of converting pyruvic acid or pyruvate to malic acid or malate, or vice versa, a recombinant vector containing the gene, a microorganism transformed with the recombinant vector, and a method for preparing succinic acid using the transformed microorganism.

A method for constructing an ALA production bacterial strain, the method enhances the activity of related enzymes promoting the synthesis of oxaloacetate and in the 5-aminolevulinic acid (ALA) production bacterial strain, or introducing exogenous related enzymes promoting the synthesis of oxaloacetate, such as phosphoenolpyruvate carboxylase or pyruvate carboxylase, and/or reducing the activity of related enzymes in the downstream metabolic pathway of succinyl coenzyme A in the bacterial strain, such as succinyl coenzyme A synthetase or succinate dehydrogenase, and/or reducing the activity of phosphoenolpyruvate carboxylated kinase and/or malic enzyme. An ALA high-yield bacterial strain constructed by utilizing the method, and method for utilizing the bacterial strain to prepare ALA.

The invention provides a method for improving the microbial production of 1,3-propanediol by constructing a gene engineering bacterium. The method comprises the following steps: constructing an expression vector with the inserted malic enzyme gene; delivering the expression vector in host bacteria generating 1,3-propanediol; adding an inducer to induce the overexpression of the malic enzyme gene in the fermentation and culture process; and adopting the aerobic fermentation means and performing the fed batch of substrate glycerol to produce 1,3-propanediol. The method is characterized in that the constructed gene engineering bacterium can express more malic enzyme than the original strain in the fermentation process, thus the convertion from pyruvic acid to malic acid can be promoted, the circulation of tricarboxylic acid can be promoted, the bacterium can generate more nicotinamide adenine dinueleotide (NADH) and energy (ATP), the activity of 1,3-propanediol oxidation-reduction enzymein the bacterium and the glycerol conversion rate can be increased. The invention has the following advantages: the substrate glycerol utilization rate of the producing bacterium can be increased, the concentration and production strength of the fermented 1,3-propanediol can be obviously increased, the yield of 1,3-propanediol can be increased and the production cost can be reduced.

The invention relates to a rhodotorula glutinis oil genetic engineering strain and a construction method and an application thereof. The construction method of the genetic engineering strain is mainly as follows: utilizing rDNA (recombinant deoxyribonucleic acid) of rhodotorula glutinis as a target sequence for homologous integration, using strong promoter genes PGK1 of saccharomyces cerevisiae and malate dehydrogenase genes ME of chaetomium cochloides to construct an expression vector to be introduced into rhodotorula glutinis, and enabling ME genes to obtain high-efficient expression in a rhodotorula glutinis body, wherein the content of lipid in a transformant is improved by 2.5 times in comparison with a wild strain. According to the construction method disclosed by the invention, key enzyme genes and a strong promoter for anabolism of the lipid are introduced on the basis that the anabolism of microbial oil is known, so that the lipid metabolism is regulated and controlled, and the yield of oil is improved. The genetic engineering strain can be applied to production of the microbial oil and development of functional oil related products, such as medicaments, health care products and the like.

A method for producing gamma-linolenic acid oil by microbial fermentation comprises the following process steps: culturing high-level expression strains of malic enzyme and gamma-linolenic acid synthetase; performing activation and amplification culture on original strains; then performing primary fermentation, secondary fermentation, third-stage fermentation and a fourth-stage fermentation; dehydrating, drying and extracting the oil. The method has the advantages of good strains, high yield, high dry bacteria yield up to 10-30%, good oil yield up to 35-55%, satisfactory gamma-linolenic acid yield up to 18-29% and low production cost. The method can help reduce the cost of raw materials for the production process by 50-70%. The semi-continuous fermentation or continuous fermentation process is advanced, scientific and mature, can be used for large scale industrialized production, and achieve 50-500t fourth-stage fermentation level.

A bacterium belonging to the genus Escherichia which has an ability to produce L-lysine or L-threonine and which is modified so that a malic enzyme does not function normally in a cell, and a method for producing L-lysine or L-threonine, comprising culturing the bacterium in a medium to produce and cause accumulation of L-lysine or L-threonine, and collecting the L-lysine or L-threonine from the medium.

The invention relates to a method for promoting synthesis of DHA in schizochytrium limacinum grease, and particularly relates to a method for prompting synthesis of docosahexaenoic acid (DHA) in schizochytrium limacinum grease. The method is characterized in that the activity of malic enzyme in a tricarboxylic acid transport system is inhibited by adding an exogenous control factor, so that the content of the NADPH in the schizochytrium limacinum can be greatly reduced, the synthesis of a great amount of DHA can be promoted, and the exogenous control factor is sesamol. According to the method, a fermentable carbon source also can be selected to reduce metabolic flux of a pentose phosphate pathway in the bacteria, so that the content of NADPH in the schizochytrium limacinum can be greatly reduced, the synthesis of a great amount of DHA can be promoted, and the largest content of the DHA in the grease of final bacteria can reach 55.3 percent. The method is simple, easy, obvious in effect, capable of greatly increasing the content of DHA in the bacteria grease and is likely to realize the industrialized application.

The invention discloses a nucleotide analogue and a synthesis and a application thereof. The constitutional formula of the nucleotide analogue is that: Tetra-O-Acetyl-D-Ribose is adopted as a raw material and generates the nucleotide analogue containing a triazole ring formwork through four steps of reaction, wherein R is C2-C15 saturated or unsaturated alkyl, or C2-C10 saturated or unsaturated alkyl containing hetero atoms; or R is R1 which is hydroxyl, alkoxy, amino or substituent amido; or R is R2 which is aryl or saturated or unsaturated alkyl; or R is R3 which is halogen, amono, methoxyl, methyl or other substituent. The nucleotide analogue can restrain the growth of microorganism such as escherichia coli, and can be used as an inhibitor of dehydrogenase such as malate dehydrogenase.

The invention discloses a reduction method of a NAD analog and an application of the reduction method. According to the method, a reducing agent is a formic acid compound, a catalyst is hydrogenlyasewhich can use the formic acid compound, and when the hydrogenlyase is used for oxidating the formic acid compound, the NAD analog is converted into a reducing state. The method can be used for producing the reducing state NAD analog or a deuterated reducing state analog, and can also be used for providing reducing power for an enzyme reaction consuming the reducing state NAD analog, and the reducing state NAD analog can also be used as coenzyme to be used for a reduction reaction of enzyme catalysis of malic enzyme ME-L310R/Q401C, D-lactate dehydrogenase DLDH-V152R, saccharomyces cerevisiae alcohol dehydrogenase and the like. Wide application of the NAD analog is facilitated. According to the reducing method of the NAD analog, under the mild condition, the regenerated reducing state NAD analog can be used for preparing malic acid or lactic acid, and can also be used as oxidation-reduction power for adjusting and controlling the metabolism intensity of the malic acid or the lactic acidin microorganisms.

The invention discloses a method for reducing NAD analogues by utilizing methanol and applications. In the method, a reducing agent is the methanol, a catalyst is the methanol dehydrogenase capable ofutilizing the methanol, and NAD analogues can be converted into the reduced states of the NAD analogues while the methanol dehydrogenase oxidizes the methanol; the method can be used for producing the reduced state NAD analogues or the reduced state analogues of deuteration and can provide reduced state coenzymes for the enzymatic reaction consuming the reduced state NAD analogues; the reduced state NAD analogues can be used as coenzymes to be applied to the reduction reaction catalyzed by enzymes such as malic enzyme ME-L310R/Q401C, D-lactic dehydrogenase DLDH-V152R and saccharomyces cerevisiae alcohol dehydrogenase, so that the wide application of the NAD analogues can be realized; the reduction method of the NAD analogues can be performed under a mild condition; and the regenerated reduced state NAD analogues can be used for preparing malic acid or lactic acid and can be used as redox power to control and regulate the metabolic intensity of the malic acid or lactic acid in microorganisms.

The invention discloses a gene SgME1 which codes malic enzyme as well as an application of the gene SgME1. The nucleotide sequence of the malic enzyme gene SgME1 is shown as SEQIDNO:1, and the amino acid sequence of coded proteins is shown as SEQIDNO:2. The SgME1 gene has the functions of regulating and controlling synthesis of malic acid of stylosanthes guianensis, and over-expressing the SgME1 to accelerate synthesis of malic acid of yeasts, hairy roots of beans and Arabidopsis and increasing the aluminum tolerance of the SgME1.

The present disclosure provides a method of determining treatment for cancer comprising identifying the absence of malic enzyme 1 (ME1) and treating with an inducer of ferroptosis.

The present invention relates to methods, compositions, and diagnostic tests for treating and diagnosing cancer and other related diseases that result in dysregulation of malic enzyme 2. In particular, the methods and compositions include combination therapy, such as with a combination of two or more ME2 inhibitors or a combination of an ME2 inhibitor and an anticancer agent.

Lipogenic yeasts bioengineered to overexpress genes for lipid production, and methods of use thereof. The yeasts are modified to express, constitutively express, or overexpress an acetyl-CoA carboxylase, an alpha-amylase, an ATP citrate lyase, a diacylglycerol acyltransferase, a fatty acid synthase, a glycerol kinase, a 6-phosphogluconate dehydrogenase, a glycerol-3-phosphate dehydrogenase, a malic enzyme, a fatty acyl-CoA reductase, a delta-9 acyl-CoA desaturase, a glycerol-3-phosphate acyltransferase, a lysophosphatidate acyltransferase, a glucose-6-phosphate dehydrogenase, a beta-glucosidase, a hexose transporter, a glycerol transporter, a glycoside hydrolase enzyme, an auxiliary activity family 9 enzyme, or combinations thereof. The yeasts in some cases are also modified to reduce or ablate activity of certain proteins. The methods include cultivating the yeast to convert low value soluble organic stillage byproducts into lipids suitable for biodiesel production and other higher value uses.

The invention relates to a recombinant bacterium for coordinately expressing glucose-6-phosphate dehydrogenase (G6PD)2 and malic enzyme (ME)2 in mortierella alpine, and further relates to a constructing method of the strain and an application of the strain. According to the recombinant bacterium, the constructing method and the application, the high-arachidonic-acid-yield mortierella alpine gene recombinant strain is constructed with mortierella alpine uracil auxotroph as a material; the important effects of the G6PD2 and the ME2 in the fatty acid synthesis process and the fatty acid desaturation process are verified; due to coordinate and ectopic expression of the two genes, the effects of increasing the yield and the desaturation degree of fatty acid are effectively achieved at the same time, the proportion of ARA in the total fatty acid is remarkably increased, fermentation time is effectively shortened, and the recombinant bacterium is of great significance in lipid synthesis basic principle researching and product development of the oil-producing fungus in mortierella alpine ATCC 32222.

The invention discloses a malic enzyme (NADP-ME) gene and application thereof. The novel NADP-ME gene provided by the invention can be stably expressed and inherited in wheat, thus obviously improving photosynthetic efficiency compared with an acceptor material. Therefore, the NADP-ME gene provides a significant gene source and parent material support for improving photosynthetic efficiencies of C3 crops through a C4 high-photosynthetic efficiency gene and for selecting a high-photosynthetic efficiency transgenic wheat variety which substantially improves yield level.

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.

Transgenic Yarrowia species are disclosed herein that comprise a polynucleotide encoding a cytosolic malic enzyme, a lipid content that is at least about 35% by weight of the dry cell weight of the Yarrowia species, and an engineered polyunsaturated fatty acid (PUFA) biosynthetic pathway, wherein overexpression of the cytosolic malic enzyme increases lipid content.

The invention relates to a genetic biochemical site marker for cerebral ischemia inbred line meriones unguiculatus and application of the biochemical site marker. The genetic biochemical site marker for the cerebral ischemia inbred line meriones is selected from at least one of glucose-6-phosphate dehydrogenase-1, esterase-3, carbonic anhydrase-2, alkaline phosphatase-1, isocitrate dehydrogenase-1, malic enzyme-1, glucose phosphate translocase-1, renal catalase-2, peptidase-3, glucose phosphate isomerase-1, hemoglobin-beta chain, transferring, esterase-1, esterase-10, glycerol phosphate dehydrogenase-1, beta-glucuronidase-1, esterase-2, lactic dehydrogenase regulator-1, serum protein-1, amylase-1, esterase-6, esterase-8, esterase-9, esterase-4, catalase-1 and esterase-12.

The invention discloses a recombinant vector over-expressing a malic enzyme gene, and its application. The recombinant vector comprises a screening labeled gene expression cassette and a target gene expression cassette; the screening labeled gene expression cassette comprises a resistance screening gene, the upstream of the resistance screening gene is fused with an Escherichia coli P1 promoter, and the downstream of the resistance screening gene is fused with an Escherichia coli ccdB terminator; and the target gene expression cassette comprises a Phaeodactylum tricornutum fcpC gene promoter, the malic enzyme gene, a Phaeodactylum tricornutum fcpA gene terminator and an Omega leader sequence, and the Omega leader sequence is in the downstream of the Phaeodactylum tricornutum fcpC gene promoter. The recombinant vector is small, is suitable for converting, is easy to operate, is a homologous sequence of the Phaeodactylum tricornutum, and benefits for expression. The over-expression of the malic enzyme gene can substantially improve the content of lipids in micro-algal cells, and lays the foundation for the researches of biological energy.