72 results about "Phosphoenolpyruvate carboxykinase" patented technology

The invention relates to isolated nucleotide sequences from Coryneform bacteria which code for the pck gene encoding the enzyme phosphoenol pyruvate carboxykinase (PEP carboxykinase). The invention also relates a process for the fermentative preparation of L-amino acids, in particular L-lysine, L-threonine, and L-glutamate by attenuation of the pck gene.

The present invention relates a method for transforming a rice plant to provide it with a C4 photosynthetic pathway by way of the introduction of some genes participating in a C4 photosynthetic pathway. To this end, the method of the invention comprises introducing a phosphoenolpyruvate carboxylase (PEPC) and a gene coding for a phosphoenolpyruvate carboxykinase (PCK) which has been connected with a DNA fragment coding for a transit peptide into a rice plant.

The invention provides a gene engineering bacterial strain for producing L-malic acid as well as a construction method and application thereof. In the invention, key phosphoenolpyruvate carboxykinase(PEPCK) related to the L-malic acid, malate dehydrogenase (MDH) and fumarase (FumC) are transformed to obtain the gene engineering bacterial strain; a deactivation mutation is carried out on the fumCto cause the interruption of a metabolic flux at the converting position from the malic acid to the fumaric acid so as to accumulate the target product L-malic acid; before the mdh, the promoter of aninner source pepck gene is added to generate the metabolic flux biased towards the malic acid; in addition, through the inducement of monofluorine sodium acetate, the activity of the PEPCK is improved, and the product feedback inhibition is greatly reduced. The bacterial strain applied to the fermentative production of the L-malic acid so as to obviously improve the yield of L-malic acid. The invention has advantages of simple technology, obvious effect and low cost and can satisfy the demand of markets.

The invention relates to isolated nucleotide sequences from Coryneform bacteria which code for the pck gene encoding the enzyme phosphoenol pyruvate carboxykinase (PEP carboxykinase). The invention also relates a process for the fermentative preparation of L-amino acids, in particular L-lysine, L-threonine, and L-glutamate by attenuation of the pck gene.

This invention comprises an extract of Artemisia dracunculus that can be used for the treatment and prevention of diabetes, diabetic complications, metabolic syndrome and other comorbidities that share the underlying commonality of insulin resistance. The invention includes the identity of six compounds from the extract that contribute to the activity of the extract by inhibiting protein tyrosine phosphatase-1B (PTP-1B) activity, phosphoenolpyruvate carboxykinase (PEPCK) gene expression or aldose reductase activity (ALR2). The compounds include 4,5-Di-O-caffeoylquinic acid, davidigenin, 6-demethoxycapillarisin, 2′,4-dihydroxy-4′-methoxydihydrochalcone, 2′,4′-dihydroxy-4-methoxdihydrochalcone and sakuranetin.

The invention discloses recombinant bacteria for increasing the yield of succinic acid and a construction method thereof. The recombinant bacteria disclosed by the invention are obtained so as to increase the enzyme activities of phosphoenolpyruvate carboxylase (PPC) and phosphoenolpyruvate carboxykinase (PCK) in escherichia coli or mutant strains of the escherichia coli. Experiments prove that the expressions of ppc and pck genes of the E.coli are respectively regulated and controlled by using constitutive controlling elements with different expression intensity, and the law between the enzyme activities of the PPC and the PCK and the succinic acid production is explored; and on the basis, the PPC and PCK catalyzing enzymes are simultaneously used for giving play to respective catalyzing advantages through coordinated regulation of the ppc and pck genes of the E.coli, so that the yield and the conversion rate of the E.coli succinic acid are obviously increased.

The invention discloses construction and applications of a corynebacterium glutamicum mutant strain for producing L-homoserine, and belongs to the technical field of fermentation engineering. Corynebacterium glutamicum ATCC 13032 is taken as a starting strain to knock out regulatory protein McbR, homoserine kinase, transport protein MetD, phosphoenolpyruvate carboxykinase; the expression of isocitrate dehydrogenase is down regulated; transport protein BrnFE, aspartic semialdehyde dehydrogenase and homoserine dehydrogenase are overexpressed; and the expression of aspartate kinase, pyruvate carboxylase and the aspartate kinase I derived from escherichia coli are enhanced. Shake flask culture can be performed on the mutant strain for 48 h, and the yield of L-homoserine can reach 8.8 g / L.

The invention discloses a corynebacterium glutamicum recombinant strain for producing 5-aminolevulinic acid. An establishing method comprises steps as follows: (1) a lactic dehydrogenase encoding gene ldhA and acetic acid producing genes pta-ackA, pqo and cat are knocked out from corynebacterium glutamicum ATCC 1303, and a strain CG4 is obtained; an sod promoter is inserted in front of a phosphoenolpyruvate carboxylase encoding gene in the CG4, and a strain CG5 is obtained; a phosphoenolpyruvate carboxykinase encoding gene pck is knocked out from the CG5, and a strain CG6 is obtained; (2) plasmids of an overexpressed 5-aminolevulinic acid synthase gene are transferred into the CG6, and a recombinant strain L is obtained; (3) 5-aminolevulinic acid transport protein plasmids are transferred into the L. The recombinant strain can promote 5-aminolevulinic acid to be transported outside corynebacterium glutamicum cells, and the yield of 5-aminolevulinic acid is 112.3% higher than that of a contrast strain.

The invention relates to a corn C4 type phosphoenolpyruvate carboxykinase gene and an application thereof in wheat production. The gene is as indicated in SEQ ID No:1 of a sequence table, a gene clone vector is pMD19-pepc, an eucaryon expression vector is p3301-pepc, host bacteria is E.coli competent cell DH5 alpha, carboxylase is has an amino acid sequence as indicated by SEQ ID No:2. In the invention, the corn C4 type phosphoenolpyruvate carboxykinase is led into the wheat, functional analysis on transgenic plants indicates that net photosynthetic rate of the transgenic plants is greatly improved compared with the net photosynthetic rate of the plants which are not transgenic, enjoying an increase of 1.39 times. The research is so far the research in which the net photosynthetic rate of C3 crop is raised to the largest degree. The result of the functional analysis of the transgenic plants indicates that the related corn C4 type high light efficiency pepc gene can greatly improve the net photosynthetic rate of C3 crops such as wheat and the like and provides important technological and material support for high light efficiency transgenic breeding of crops such as wheat and the like.

The invention discloses a DNA (Deoxyribonucleic Acid) molecule related to photosynthesis and the application thereof. The DNA molecule is formed by sequentially connecting the following DNA segments: a DNA segment 1, a DNA segment 2, a DNA segment 3, a DNA segment 4 and a DNA segment 5. The DNA segment 1 is formed by sequentially connecting a promoter for driving PEPC (phosphoenolpyruvate carboxykinase) protein expression, an encoding gene of PEPC protein and a terminator; and the DNA segment 2 is formed by sequentially connecting a promoter for driving PPDK (Pyruvate Orthophosphate Dikinase)protein expression, an encoding gene of PPDK protein and a terminator. An experiment proves that a transgenic plant obtained by guiding a plurality of C4 pathway key enzyme bases into the C3 plant has higher photosynthetic efficiency than a wild plant (a C3 plant). The invention has a certain reference for practical operation for increasing crop photosynthetic efficiency by using a multigene transformation technology from now on.

The present invention relates to a recombinant adenovirus with increased in-vivo safety, tissue specificity, and anticancer activities, and a use thereof. Specifically, the recombinant adenovirus comprising: a promoter of the liver tissue-specific phosphoenolpyruvate carboxykinase (PEPCK) gene; a trans-splicing ribozyme which is operably linked to the promoter and acts on a cancer-specific gene; a therapeutic gene or a reporter gene which is linked to the 3′ exon of the ribozyme; and a serotype 35 fiber knob and a serotype 5 shaft, in which the orf4 gene is deleted from adenovirus E1, E3 and E4 orf1, shows remarkable in-vivo safety, high specificity for a target tissue, and remarkable anticancer effects, and thus can be useful for an anticancer drug or a cancer diagnostic agent as a gene delivery vector.

The invention provides a method for improving the quality of a loquat fruit by agricultural measures, which combines the reflection film spreading in an orchard and the suspension of magnets. The invention can accurately and effectively adjust and control key enzymes for forming and degrading the main organic acid (malic acid) of the loquat fruit. At the early growth stage of the fruit, the invention improves the activity of phosphoenolpyruvate carboxykinase and NAD-malic dehydrogenase in the pulp; and at the most critical stage for the accumulation of organic acid of the fruit, the invention can promote the activity decline of the PEPC in the pulp, obviously inhibit the activity of the NAD-MDH and enables the activity of NADP-ME in the pulp, reduce the synthesis of malic acid and enhance the degradation of malic acid, thereby obviously reducing the organic acid content of the fruit of the high-acid content loquat, and simultaneously increasing the TSS content of the mature pulp. The invention can obviously reduce the titratable acid (TA) in the fruit and increase the soluble solid (TTS) content of the mature pulp and the weight of a single mature fruit, thereby obviously improving the quality of the mature loquat fruit.

The invention discloses a barnyard grass phosphoenolpyruvate carboxylase gene and coded protein and appliance, which is one ribotide sequence in the following sequences: 1) sequent graph SEQ ID No.:1;2) DNA sequence hybrid ribotide sequence limited by SEQ ID No.:1. the coded gene protein is one of SEQ ID No.:2 in the 1) sequence graph; 2) SEQ ID No.:2 in the 2) sequence graph with two amino residue sequence replaced, lost or added form one to ten to improve plant photosynthesis efficiency.

The invention provides an effective method for improving N-acetylglucosamine (GlcNAc) production by engineered B. subtilis Deletion of phosphoenolpyruvate carboxykinase encoding gene pckA and encoding pyruvate kinase gene pyK in recombinant GlcNAc-producing strain BSGNK-PxylA-glmS-P43-GNA1 (BSGNK) is first performed to enhance GlcNAc production, followed by overexpression of pyruvate carboxylase encoding gene pycA for facilitating cell growth. Finally, the GlcNAc production of the recombinant strain BPTS3 reached to 11.3 g / L, which was 1.84-fold of BSGNK. This method can be used for improve cellular property of engineered B. subtilis for GlcNAc production, which can be further applied to industrial production of GlcNAc.

Variant phosphoenolpyruvate carboxylase (PEPC) genes are described. The encoded PEPC variants contain amino acid substitutions and have altered kinetic and / or regulatory properties with respect to wild-type PEPC. A variant PEPC gene may be expressed in a plant to improve one or more traits such as CO2 assimilation rate, water use efficiency, and yield.

The invention discloses cloning and application of a maize phosphoenolpyruvate carboxykinase gene promoter. A method comprises the following steps of: cloning a phosphoenolpyruvate carboxykinase gene promoter from maize, fusing the cloned phosphoenolpyruvate carboxykinase gene promoter with a target gene coding area or a ribonucleic acid interference (RNAi) structure thereof to construct a plant expression structure, and introducing a recombinant gene into a plant cell by transgenic technology to generate a transgenic plant; and selecting the target character of the transgenic plant to obtain the transgenic plant of which the stress resistance or economic character is improved obviously or descendants thereof, so that new germplasm and species which have application prospects in plant breeding are created.

The invention discloses recombinant escherichia coli for efficiently producing succinic acid and a construction method thereof, and belongs to the technical field of bioengineering. The method comprises the following steps: knocking out a by-product encoding gene of a succinic acid producing strain E.coli FMME-N-2 to obtain a strain E.coli FMME-N-5(delta foc A-pflB-delta ldhA-delta pta-ackA); performing overexpression from actinobacillus succinogenes phosphoenolpyruvate carboxykinase pck and bacillus stutzeri phosphite dehydrogenase ptxD; introducing the constructed plasmid pTrcHisA-pck-ptxD into an expression host E.coli FMME-N-5(delta foc A-pflB-delta ldhA-delta pta-ackA), and screening through an ampicillin resistance flat plate to obtain an engineering bacterium E.coli FMME-N-5(delta foc A-pflB-delta ldhA-delta pta-ackA)-pck-ptx-D for efficiently producing succinic acid. The engineering strain adopts a two-stage fermentation strategy on a 7.5 L fermentation tank, the fermentation time is 96h, the succinic acid yield reaches 137g / L, the succinic acid yield reaches 1g / g glucose, the production strength is 1.43 g / L / h, byproducts lactic acid and formic acid are not accumulated, acetic acid is 1-2g / L, and the engineering strain has certain industrial production potential.

The invention relates to a kit for diagnosing / measuring lithium by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay, wherein, the activity of the kit can be inhibited by lithium. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of lithium, and belongs to the technical field of medical / industrial / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, magnesium chloride, inositol-1-phosphate, an oxaloacetic acid, a pyruvic acid, inositol-1-phosphatase, phosphoenolpyruvate carboxykinase, malic dehydrogenase and a stabilizer. Through mixing a check sample and a lithium sample respectively with the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, the degree / velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, and the difference in the degree / velocity of the decrease between the check sample and lithium sample is compared, thereby measuring the concentration of lithium.

The invention relates to a kit for diagnosing / measuring carbon dioxide by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the carbon dioxide, and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, a phosphoenolpyruvic acid, phosphoenolpyruvate carboxykinase, malic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the degree of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of the carbon dioxide.

The invention belongs to the field of biotechnology, and discloses a human replication-deficient recombinant adenovirus vector for efficiently expressing leishmania phosphoenolpyruvate carboxykinase (PEPCK) aiming at the current situation of leishmania. Specifically, carrying out homologous recombination on a recombinant vector pShuttle-CMV-Leish-PEPCK constructed by a modified PEPCK gene and a pShuttle-CMV vector and a pAdEasy-1 vector, carrying out PacI cutting on the obtained plasmid vector pAd5-L.m-PEPCK, and transfecting into a human embryo kidney cell 293, so as to obtain the replication-deficient recombinant adenovirus vector Ad5-L.m-PEPCK. Through Coomassie brilliant blue R250 dyeing and Western blot analysis of a specific anti-His tag antibody, it is shown that the recombinant adenovirus vector Ad5-L.m-PEPCK can efficiently express the PEPCK protein. As the leishmania phosphoenolpyruvate carboxykinase (PEPCK) is the most immunodominant leishmania disease antigen which is found recently. Therefore, the recombinant adenovirus vector Ad5-L.m-PEPCK disclosed by the invention can be used for a vaccine for the leishmaniasis, so that the prevention and control of the leishmaniasis are facilitated.

The invention relates to an inorganic phosphorus (phosphate radical) diagnosis / determination kit using the technology of an enzyme colorimetric method and an ELISA method, a method for determining inorganic phosphorus (phosphate radical) concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food, and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, adenosine diphosphate, oxaloacetic acid, ammonium chloride, potassium chloride, phosphoenolpyruvate carboxykinase, pyruvate kinase, pyruvate dehydrogenase and a stabilizer. The method comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV / visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the inorganic phosphorus (phosphate radical) concentration.

The invention discloses a Yinhong vitis vinifera phosphoenolpyruvate carboxykinase PEPCK gene, a primer for amplifying the Yinhong vitis vinifera phosphoenolpyruvate carboxykinase PEPCK gene, a primerfor fluorescent quantitative RT-qPCR of the Yinhong vitis vinifera phosphoenolpyruvate carboxykinase PEPCK gene and an application of the Yinhong vitis vinifera phosphoenolpyruvate carboxykinase PEPCK gene to regulation and control of sugar acid content of Yinhong vitis vinifera fruits. SPSS software analysis shows that the expression quantity of the PEPCK gene is in extremely remarkable positivecorrelation with accumulation of glucose and fructose in the color changing period, besides, the expression quantity of the PEPCK gene is in remarkable negative correlation with the content change offruit organic acid, and it can be proved that the PEPCK gene plays a role in regulating and controlling synthesis of saccharic acid in the Yinhong vitis vinifera fruit ripening process. The inventionlays a foundation for subsequent researches on exact functions of the Yinhong vitis vinifera PEPCK gene, sugar acid metabolism regulation ways in which the Yinhong vitis vinifera PEPCK gene participates and the like.

The invention provides an effective method for improving N-acetylglucosamine (GlcNAc) production by engineered B. subtilis Deletion of phosphoenolpyruvate carboxykinase encoding gene pckA and encoding pyruvate kinase gene pyK in recombinant GlcNAc-producing strain BSGNK-PxylA-glmS-P43-GNA1 (BSGNK) is first performed to enhance GlcNAc production, followed by overexpression of pyruvate carboxylase encoding gene pycA for facilitating cell growth. Finally, the GlcNAc production of the recombinant strain BPTS3 reached to 11.3 g / L, which was 1.84-fold of BSGNK. This method can be used for improve cellular property of engineered B. subtilis for GlcNAc production, which can be further applied to industrial production of GlcNAc.

The invention relates to the nucleic acid molecule and method for improving C3 plant photosynthesis by utilizing transgenic technology, wherein the nucleic acid molecule is the nucleic acid molecule which encodes sugar cane phosphoenolpyruvate pyruvate carboxylase (PEPC), and the method consists of leading the enzyme-sugar cane phosphoenolpyruvate pyruvate carboxylase (PEPC) participating C4 photosynthesis channels into plants having C3 type photosynthesis channels such as rice, so as to provided them with C4 photosynthesis channels, and make them have higher efficiency of photosynthetic activity. The invention also relates to the expression vectors for converting the C3 plants, and the use of the method.

The invention discloses a transgene carrier for changing the mouse movement ability and the metabolic function by the specific expression of PEPCK-C (phosphoenolpyruvate carboxykinase-cytosolic) in the muscular tissue. The transgene carrier comprises the promoter of the gene of the 3.3kb alpha-skeletal actin of a mouse, the cDNA (deoxyribonucleic acid) of the PEPCK of the mouse and the 3'-end no-translation area of a bovine growth hormone gene (bGH); the transgene carrier is provided with a resistance gene-puromycin gene used for screening in mammalian cells, and a resistance gene-ampicillin gene used for screening in prokaryotic cells. The transgene carrier disclosed by the invention can be used for cloning a somatic cell so as to construct a PEPCK-Cmus transgene mouse.

The invention discloses a genetically modified microorganism capable of producing 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and / or hexanedioic acid at a high yield, and a production method for 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and / or hexanedioic acid using the genetically modified microorganism. The genetically modified microorganism has the ability to produce 3-hydroxyhexanedioic acid, (E)-hex-2-enedioic acid and / or hexanedioic acid, and disrupts the function of pyruvate kinase and enhances the activity of phosphoenolpyruvate carboxykinase and the activity of enzymes that catalyze a reaction for generating 3-hydroxyadipyl-CoA by reducing 3-oxoadipyl-CoA.

Provided herein are methods, compounds, and compositions for reducing expression of phosphoenolpyruvate carboxykinase-mitochondrial (PEPCK-M) mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions for preventing or decreasing diabetes, obesity, metabolic syndrome, diabetic dyslipidemia, and / or hypertriglyceridemia in an animal. Such methods, compounds, and compositions are useful to treat, prevent, delay, or ameliorate any one or more of diabetes, obesity, metabolic syndrome, diabetic dyslipidemia, and / or hypertriglyceridemia, or a symptom thereof.

The present invention relates to a recombinant adenovirus with increased in-vivo safety, tissue specificity, and anticancer activities, and a use thereof. Specifically, the recombinant adenovirus comprising: a promoter of the liver tissue-specific phosphoenolpyruvate carboxykinase (PEPCK) gene; a trans-splicing ribozyme which is operably linked to the promoter and acts on a cancer-specific gene; a therapeutic gene or a reporter gene which is linked to the 3′ exon of the ribozyme; and a serotype 35 fiber knob and a serotype 5 shaft, in which the orf4 gene is deleted from adenovirus E1, E3 and E4 orf1, shows remarkable in-vivo safety, high specificity for a target tissue, and remarkable anticancer effects, and thus can be useful for an anticancer drug or a cancer diagnostic agent as a gene delivery vector.

The invention discloses a detection method and diagnosis reagent box of carbon dioxide (bicarbonate) in the medical detection domain, which comprises the following parts: buffer liquid, phosphoenolpyruvate, muscle nucleoside, phosphoenolpyruvate carboxylase, nucleoside phosphatase, xanthine oxidase, alcohol, hydrogen peroxidase, oxidized coenzyme, aldehydo dehydrogenase and stabilizer. The detection method comprises the following steps: mixing the sample and reagent at certain bulk proportion to do enzyme coupling reaction; setting the final reactant under the biochemical analyzer; detecting main wavelength absorbance variation condition (speed); counting the content of carbon dioxide (bicarbonate).