65 results about "Pentose phosphate pathway" patented technology

The invention relates to the methods of manufacturing five-carbon sugars and sugar alcohols as well as other compounds derived from pentose-phosphate pathway from readily available substrates such a hexoses using metabolically engineered microbial hosts.

The invention belongs to the field of microorganisms and particularly relates to Corynebacterium glutamicum and its construction method and application. Serine at the 361th site of a gnd gene for coding 6-phosphogluconate dehydrogenase in the cell of Corynebacterium glutamicum is mutated into phenylalanine. Corynebacterium glutamicum is used for point mutation of the gnd gene in the pentose phosphate pathway so that serine at the 361th site of 6-phosphogluconate dehydrogenase is replaced by phenylalanine and thus allosteric regulation of 6-phosphogluconate dehydrogenase is reduced, the ability of NADPH regeneration is improved, enough NADPH is supplied for proline synthesis and a proline yield is improved. An experiment result shows that the Corynebacterium glutamicum is an L-proline high-yield strain, effectively accumulates L-proline, improves an L-proline yield and lays the foundation for the industrial production of L-proline.

The present invention provides a method of preparing cyclic adenosine monophosphate, which includes the use of Arthrobacter sp. as an engineering bacterium, and the addition of precursors, inhibitors of glycolytic pathway and/or activators of tricarboxylic acid cycle into a fermentation medium, so as to acquire the cyclic adenosine monophosphate by fermentation. Further provided is a use of precursors, inhibitors of glycolytic pathway and/or activators of tricarboxylic acid cycle for preparing cyclic adenosine monophosphate by Arthrobacter sp.. The present invention inhibits the excessive glycolysis, increases the flux of pentose phosphate pathway, and enhances the tricarboxylic acid cycle by the addition of precursors, inhibitors of glycolytic pathway and/or activators of tricarboxylic acid cycle, so as to redistribute rationally the metabolic flux of central metabolic pathways, make the carbon flux more efficiently direct to the target product - cyclic adenosine monophosphate, and synthesize cyclic adenosine monophosphate by using carbon source efficiently.

The invention discloses tetrahydropyridine high-yield Corynebacterium glutamicum and application thereof and belongs to the field of bioengineering. Feedback inhibition is relieved through site-directed mutagenesis of Corynebacterium glutamicum aspartokinase gene lysC, and expression of mutational LysC is intensified through promoter replacement. Further, pentose phosphate pathway of host bacteriais further intensified through promoter replacement to meet needs on reducing power NADPH in efficient synthesis of tetrahydropyridine. Finally, tetrahydropyridine high-yield Corynebacterium glutamicum is obtained by transferring tetrahydropyridine synthetic gene cluster ectABC of P. stutzeri into recombinant bacteria. By recombinant Corynebacterium glutamicum, tetrahydropyridine can be efficiently synthesized by utilizing cheap raw materials like glucose and maize plasm; compared with recombinant Escherichia coli produced strains, tetrahydropyridine high-yield Corynebacterium glutamicum hasbetter bio-safety and is of great significance to industrial production and large-scale application of tetrahydropyridine.

Coordinately regulated over-expression of the genes encoding glucose 6-phosphate dehydrogenase [“G6PDH”] and 6-phospho-gluconolactonase [“6PGL”] in transgenic strains of the oleaginous yeast, Yarrowia lipolytica, comprising a functional polyunsaturated fatty acid [“PUFA”] biosynthetic pathway, resulted in increased production of PUFAs and increased total lipid content in the Yarrowia cells. This is achieved by increased cellular availability of the reduced form of nicotinamide adenine dinucleotide phosphate [“NADPH”], an important reducing equivalent for reductive biosynthetic reactions, within the transgenic microorganism.

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.

An object of the present invention is to provide a method for producing ethanol efficiently even in the presence of a fermentation inhibitor in a saccharified biomass. The present invention provides a method for producing ethanol from biomass, comprising: culturing a transformed xylose-utilizing yeast to overexpress the gene for at least one pentose phosphate pathway metabolic enzyme, with a saccharified biomass.

The present invention relates to a strain producing L-tryptophan and use thereof in fermentation. Specifically, the L-tryptophan producing capability of the strain is greatly improved through attenuating or blocking an oxidation stage of a pentose phosphate pathway of engineering bacteria (such as Escherichia coli or Corynebacterium glutamicum) with a genetic engineering method.

The invention relates to the field of producing succinate by E. coli fermentation. Specifically, the invention provides an engineered recombinant E. coli for producing succinate, wherein said E. coli contains one or more of the following modifications: a) enhanced activity of the protein(s) encoded by the gene(s) involved in pentose phosphate pathway (PPP), b) enhanced activity of the protein encoded by sthA gene, and optionally c) mutant lpdA gene. The invention also relates to use of the engineered recombinant E. coli for producing succinate, and a method of using the engineered recombinant E. coli for producing succinate.

Method for preventing or treating overweight or obesity, the method comprising administering to a patient in need thereof an antagonist that inhibits gene expression or activity of an enzyme that is involved or related to the pentose phosphate pathway. Preferably, the enzyme is a transketolase, and the antagonist is an antibody, an antisense molecule, an siRNA molecule, a molecule for forming a triplex nucleic acid molecule with the enzyme-encoding polynucleotide. Also disclosed are pharmaceutical compositions comprising the same, and method for screening a substance that inhibits gene expression of a PPP enzyme or its function.

The present invention relates to a method of producing methionine in Coryneform bacteria in which enzymes of the pentose phosphate pathway are over-expressed. The present invention also relates to Coryneform bacteria for producing methionine in which at least two enzymes of the pentose phosphate pathway are over-expressed.

The invention discloses corynebacterium glutamicum and corynebacterium crenatum, as well as a method for improving the yield of arginine by utilizing corynebacterium glutamicum and corynebacterium crenatum. According to the method, dtsR1 genes of corynebacterium glutamicum and corynebacterium crenatum are knocked out; then the transcriptional level of alpha-ketoglutarate dehydrogenase complex (oxoglutarate dehydrogenase complex, ODHC) regulated by the dtsR1 genes is greatly lowered, and alpha-ketoglutaric acid flows to arginine; meanwhile, Tween-80, oleic acid or a mixture of Tween-80 and oleic acid is added to a fermentation medium, so as to make up for nutrition loss caused by loss of the dtsR1 genes, and at the early logarithmic growth stage during the fermentation culturing, Tween-40 is added to induce the transcription level of the genes in the pentose phosphate pathway to be greatly improved. Thus, sufficient NADPH is provided for the synthesis of arginine, and other complicated molecular breeding methods for improving supply of NADPH and other accumulation methods for improving content of alpha-ketoglutaric acid in the tricarboxylic acid cycle pathway are eliminated.

The present invention describes a recombinant yeast cell functionally expressing one or more heterologous nucleic acid sequences encoding for ribulose-1,5-phosphate carboxylase/oxygenase (EC4.1.1.39; Rubisco), and optionally one or more molecular chaperones for Rubisco, and one or more phosphoribulokinase (EC2.7.1.19; PRK), wherein one or more genes of the non-oxidative branch of the pentose phosphate pathway are overexpressed and/or wherein said yeast cell comprises a deletion or disruption of a glycerol-3-phosphate dehydrogenase (GPD) gene.

The invention discloses a method for quantitative determination of regeneration capability of 1,5-ribulose diphosphate. The method comprises the following steps: selecting the second, the third and the fourth completely spread leaves, performing non-destructive determination on apparent photosynthesis rate, taking a mean value of three leaves to represent the apparent photosynthesis rate PN of the plant; then taking the leave which has measured apparent photosynthesis rate, shearing the leave and fully mixing the leave, taking 0.1 g of the material for enzyme liquid extraction; respectively determining the phosphofructokinase vitality and glucose-6-phosphate dehydrogenase enzyme vitality expressed by the change value at unit volume and unit time in the extracted enzyme liquid, calculating phosphopentose pathway proportion in the glycometabolism according to a formula, and then multiplying by the apparent photosynthesis rate of the plant leave to obtain the regeneration capability of 1,5-ribulose diphosphate. The method can realize rapid quantitative determination of the regeneration capability of 1,5-ribulose diphosphate, and has the advantages of less step, simple calculation, and comparability and reliability of the determination result.

The invention belongs to the field of gene engineering, and discloses a construction method of a synthetic path for generating glutamic acid by utilizing xylose in corynebacterium glutamicum. The construction method comprises the following steps of (1) expressing heterogenous pentose transport protein in the corynebacterium glutamicum; (2) expressing heterogenous xylose isomerase and xylulokinasein the corynebacterium glutamicum; (3) generating alpha-ketoglutarate from generated 5-xylulosephosphate through a phosphopentose pathway and a glycolysis pathway, and carrying out weakened expressionon alpha-ketoglutarate dehydrogenase, so that more alpha-ketoglutarate flows to the synthesis of glutamic acid; and (4) modifying glutamic acid secretory protein to ensure that the corynebacterium glutamicum does not respond to external biotin pressure any more, and continuously secreting the glutamic acid out of cells. According to the constructed synthetic path from the xylose to the glutamic acid, the corynebacterium glutamicum can utilize the xylose to produce the glutamic acid in a conventional synthetic culture medium, and the synthesis from the xylose to the glutamic acid can also be conducted in a high-biotin environment such as lignocellulose.

To provide an anti-cancer agent sensitivity determination marker, which marker can determine whether or not the patient has a therapeutic response to the anti-cancer agent, and novel cancer therapeutic means employing the marker.

The anti-cancer agent sensitivity determination marker, the anti-cancer agent including oxaliplatin or a salt thereof and fluorouracil or a salt thereof, contains one or more substances selected from among an amino-acid-metabolism-related substance, a nucleic-acid-metabolism-related substance, a substance in the pentose phosphate pathway, a substance in the glycolytic pathway, a substance in the TCA cycle, a polyamine-metabolism-related substance, 7,8-dihydrobiopterin, 6-phosphogluconic acid, butyric acid, triethanolamine, 1-methylnicotinamide, NADH, NAD⁺, and a substance involved in the metabolism of any of these substances.

The invention relates to the field of gene engineering, in particular to a method for improving cellulase expression capacity of trichoderma reesei by regulating cell metabolism. The 6-phosphogluconate dehydrogenase gene 72685 of a pentose phosphate pathway of trichoderma reesei is subjected to overexpression, overexpression plasmids of the gene are constructed, and the overexpression plasmids are used for converting the trichoderma reesei, so that the protein expression quantity and the cellulase activity of the trichoderma reesei are improved.

Disclosed herein is a newly discovered problem and solution for engineering S. cerevisiae to ferment xylose to make ethanol utilizing xylose isomerase to convert xylose to xylulose for entry, via xylulokinase, into the pentose phosphate pathway. When grown on a media containing xylose xylitol tends to accumulate in the cell despite the absence of xylose reductase activity in S. cerevisiae. Xylitol inhibits the activity of xylose isomerases. One solution described is to simultaneously express an exogenous xylitol dehydrogenase along with the exogenous xylose isomerase while optionally also overexpressing xylulokinase in the absence of expression of a xylose reductase. Another solution is a xylose isomerase from Bacteroides fragilis which is less inhibited by xylitol than other xylose isomerases, exemplified by E. coli xylose isomerase. Expression of the Bacteroides fragilis xylose isomerase may be used alone, or in combination with expression of axylitol dehydrogenase and optionally over expression of xylulokinase to improve ethanol production from xylose.

The present invention provides a method for overexpressing the zwf2 gene to increase the yield of daptomycin and its application, and clones a key enzyme 6-phosphate glucose dehydrogenase gene (zwf2) in the primary metabolic pentose phosphate pathway, using Escherichia coli-Streptomyces Shuttle plasmid expression vector construction of recombinant plasmids. The recombinant plasmid was introduced into Escherichia coli ET12567, ET12567 was co-cultured with Streptomyces roseospora, and Streptomyces roseospora was transformed by the combined transfer method, and then the transformants were screened by apramycin resistance, and the genetically engineered bacteria were further verified by PCR , thereby constructing the genetically engineered strain HP-2. The Streptomyces roseospora genetically engineered bacterium HP-2 constructed by the present invention has an 18% higher output than the starting strain, and the output of daptomycin is as high as 700mg/L, which can be directly used in the industrialized production of daptomycin, which can improve Yield reduces cost.

The invention is directed to a process for the generation of male sterility in plants comprising the steps of (a) transforming a plant cell with DNA sequences that selectively inhibit the expression of essential metabolic compounds and (b) regenerating plants from said plant cells. Cells impaired in the biosynthesis of basic metabolic compounds undergo starvation and eventually die. Such pathways include amino acid biosynthesis, nucleic acid biosynthesis and other biosynthetic pathways such as citric acid cycle, pentose phosphate pathway, fatty acid metabolism, vitamin biosynthesis that will render the cell inactive due to nutrient depletion, if one or more enzymes or proteins involved in this pathway would become inactive by using inhibitory DNA constructs.

The invention discloses a mixture of rare ginsenoside and application of the mixture. The mixture of rare ginsenoside comprises rare ginsenoside and a pentose metabolic pathway inhibitor. A rare ginsenoside monomer promotes ubiquitination and degradation of HIF-1alpha (hypoxia-inducible factor-1alpha) through proteasome; the content of the HIF-1alpha in a tumor cell is lowered, theoretically, tumor glycometabolism pathways including pentose-phosphate pathways can be inhibited, and thus the anti-tumor effect is achieved. According to the mixture of rare ginsenoside and the application of the mixture, it is innovatively achieved that the rare ginsenoside monomer and a pentose-phosphate pathway inhibitor 6-amidogen nicotinamide (6-AN) are jointly applied in vitro to act on a hepatoma cell line (HepG2), and the rare ginsenoside monomer and the pentose-phosphate pathway inhibitor 6-amidogen nicotinamide (6-AN) have an obvious synergistic anticancer effect. Therefore, rare ginsenoside and atumor glycometabolism pathway inhibitor which have the effect of regulating glycometabolism pathways are jointly applied to strengthening the anti-tumor effect of the mixture, and the mixture is usedfor therapy of malignant tumors and has a wide application prospect.