82 results about "Pyruvate kinase" patented technology

The invention belongs to the technical field of chondroitin sulfate biosynthesis, and discloses a recombinant strain for high-yield chondroitin sulfate, application of the recombinant strain and a preparation method of the chondroitin sulfate. The recombinant strain provided by the invention can be used for simultaneously expressing UDP-glucosamine isomerase, UDP-glucose dehydrogenase, chondroitin polymerase, glucosamine synthetase, phosphoglucosamine mutase, UDP-N-acetylglucosamine pyrophosphorylase, glucose-6-phosphate isomerase and glucose phosphate mutase; according to the present invention, glucose-1-phosphate uridyl transferase, ATP sulfonase, sulfotransferase, pyruvate kinase and APS kinase are adopted, and the enzyme molecules are matched with the original carbon metabolism pathway in the recombinant strain cell to jointly form the enzyme-linked pathway having the efficient CSA synthesis ability, such that the high-yield and high-purity preparation of the CSA can be achieved; the method has the advantages of simple operation, high biological safety of the obtained product, easy realization of large-scale production and the like, and has a good application prospect.

The invention provides recombinant escherichia coli for de novo synthesis of anisic acid. At least one of the following improvements is carried out on an escherichia coli starting strain: (1) overexpression of O-methyltransferase PSA-OMT1 from Pleuromyces sapidus, (2) overexpression of O-methyltransferase PSA-OMT1 from Pleuromyces sapidus, and (3) overexpression of O-methyltransferase PSA-OMT1 from Pleuromyces sapidus; the method comprises the following steps: carrying out overexpression on chorismate lyase UbiC derived from E.coli MG1655 or Providencia rustiana, and carrying out the overexpression on the chorismate lyase UbiC derived from E.coli MG1655 or Providencia rustiana; (2) carrying out overexpression on a 3-deoxy-D-arabic-heptanone acid 7-phosphate (DAHP) synthase feedback inhibition mutant AroGD146N derived from E.coli MG1655 on the basis of the step (1); and (3) overexpressing synthase PpsA derived from phosphoenolpyruvate (PEP) on the basis of the step (2), and knocking out a pykF gene for coding pyruvate kinase I. Therefore, in the fermentation process of the recombinant escherichia coli, the accumulation of the anisic acid in the fermentation liquor can be realized, the maximum anisic acid shake flask yield of the recombinant escherichia coli can reach 284.9 mg / L, and a foundation is laid for the subsequent construction of high-yield anisic acid engineering bacteria.

The invention provides a method for increasing the yield of bacterial cellulose by utilizing low-dose ultraviolet radiation, and belongs to the field of microbial fermentation. The method comprises the following steps: inoculating a fermentation culture medium with a Taonella.mepensis seed solution, carrying out fermentation treatment, and carrying out ultraviolet radiation under set conditions in the fermentation process, so as to obtain a fermentation solution containing a bacterial cellulose membrane; and taking out the bacterial cellulose membrane in the fermentation liquor, and purifying the bacterial cellulose membrane to obtain the bacterial cellulose. On the basis of an HS culture medium formula, the ultraviolet irradiation time is regulated and controlled at the same time, so that the activity of cellulose synthesis related enzymes, such as the activity of UDP-glucose pyrophosphorylase and glucose phosphate mutase, is improved, the activity of glucose decomposition pathway related enzymes, such as pyruvate kinase and phosphofructokinase, is inhibited, and the activity of cellulose synthesis related enzymes is improved. The yield of the bacterial cellulose is further improved.

The invention provides a construction method and application of recombinant yarrowia lipolytica with high yield of tyrosol, and belongs to the field of bioengineering. The recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain, and the recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain. The recombinant yarrowia lipolytica disclosed by the invention expresses an ethanol dehydrogenase encoding gene, and simultaneously expresses any one or a combination of more of a DAHP synthase mutant encoding gene, a chorismate mutase mutant encoding gene, a glucose 6-phosphate dehydrogenase encoding gene, a transketolase encoding gene and a transaldolase encoding gene; meanwhile, a natural promoter of a pyruvate kinase coding gene is truncated. After the recombinant yarrowia lipolytica XJ-TL strain is subjected to fed-batch fermentation, tyrosol can be synthesized at a relatively high level. The construction method of the recombinant yarrowia lipolytica is easy to operate, and efficient synthesis of tyrosol can be achieved.

The invention provides a valine production strain and a construction method and application thereof, a designed acetohydroxy acid synthase mutant is that the 88th basic group of an ilvB gene is changed from a to c, the 382nd basic group is changed from a to g, the 413th basic group is changed from c to t, the gene sequence of a designed artificial operon comprises a promoter, an ilvB (A138V) gene or ilvB (Q30K, S128G, A138V) gene of coding mutated acetohydroxy acid synthase, and an ilvN (G20D, I21D, I21D, I21D, I21D, I21D, I21D, I21D) gene. I22F) gene, a pyk gene for coding pyruvate kinase, and a terminator; by designing a specific acetohydroxyacid synthase mutant and related biological materials and artificial operon, the strain constructed by directional modification of the strain by using a pK18mobsacB system gene editing technology based on allele exchange has the advantages of good genetic stability, high fermentation yield and the like, and valine can be stably produced.

The present invention provides a method utilizing the deletion of the regulatory protein gene tyrR1 and the overexpression of the tyrA-aroF genes to improve vanillin yield through fermentation with glucose as the substrate in Amycolatopsis. Additionally, the invention introduces a recombinant Amycolatopsis strain with a high vanillin yield, which has knocked out the regulatory protein gene tyrR1 and the pyruvate kinase gene pyk. This strain also overexpresses the endogenous chorismate mutase / prephenate dehydrogenase bifunctional enzyme gene tyrA, the 3-deoxy-7-phosphoheptulonate synthase gene aroF, and the phosphoenolpyruvate synthase gene ppsA. This recombinant Amycolatopsis ferments glucose to produce vanillin, achieving a yield of 4.72 g / L, significantly higher than that of existing technologies.

Provided herein are methods for preparing activators of Pyruvate Kinase (PK) or a salt or a hydrate thereof.

Provided are a theanine-producing strain and use thereof in tea fermentation production. A corynebacterium glutamicum is proposed, which includes an alanine decarboxylase CsAlaDC mutant. The theanine-producing strain is obtained by taking the corynebacterium glutamicum as a starting strain, knocking out in sequence an α-ketoglutarate dehydrogenase E1 subunit gene odhA, a glutamate external transporter gene Ncg11221 and a lactate dehydrogenase gene ldh; and / or expressesing a citrate synthase gene gltA, a pyruvate kinase gene pyk and a glutamate dehydrogenase gene gdh; and / or overexpressing an alanine dehydrogenase alaA and integrating a γ-glutamine synthetase GMAS into a cg1960 pseudogene locus of the corynebacterium glutamicum.

The invention relates to a compound of formula (I)which is 6-((1H-pyrazol-4-yl)sulfonyl)-2-((1-cyclopropyl-1H-pyrazol-3-yl)methyl)phthalazin-1(2H)-one; or a salt and / or solvate thereof;and to its use in treating or preventing an inflammatory disease, a disease associated with an undesirable immune response, cancer, obesity, a diabetic disease or a blood disorder.

The invention relates to the fields of genetic engineering and microbial fermentation, and discloses a recombinant strain for highly producing terpenoids as well as a construction method and application of the recombinant strain. The recombinant strain is obtained by genetic engineering modification of an original strain, compared with the original strain, the recombinant strain enhances the synthetic route of terpenoids, and the activity of pyruvate kinase, pyruvate dehydrogenase complex, glucose-6-phosphate dehydrogenase and key enzymes for synthesis of terpenoids is enhanced by using a CRISPR activation system; wherein the sgRNA expression promoters of the pyruvate kinase and the pyruvate dehydrogenase complex are respectively oleic acid induction type promoters, and the sgRNA expression promoters of the glucose-6-phosphate dehydrogenase and the terpene compound synthesis key enzyme are respectively copper ion induction type promoters. The recombinant strain can dynamically regulate and control an editing system of the terpenoids, precisely regulate and control the synthetic pathway flux of the terpenoids, and effectively improve the yield of the terpenoids (such as sclareol and bisabolol).

The invention discloses a fluorescent probe as well as a preparation method and application thereof. The fluorescent probe has a structural formula as shown in a formula I: # imgabs0 #, wherein R is selected from # imgabs1 #, and X is selected from F, Cl, Br and I. The invention provides the fluorescent probe which is novel in structure and excellent in water solubility, and the fluorescent probe can specifically recognize pyruvate kinase M2 and has the characteristic of more excellent tumor cell imaging.

The present disclosure provides a pharmaceutical composition comprising (S)-1-(5-[2H,3H-[1,4]dioxyno[2,3-b]pyridin-7-sulfonyl]-1H,2H,3H,4H,5H,6H-pyrrolo[3,4-c]pyrrole-2-yl)-3-hydroxy-2-phenylpropan-1-one (“Compound 1”) in a crystalline solid form, a spray-drying dispersion, and a solid oral administration form, and a method for preparing the same.

Methods for preparing pyruvate kinase (PK) activators or salts or hydrates thereof are provided herein. Synthetic methods for preparing pyruvate kinase (PK) activators, or salts or hydrates thereof, are provided herein. Such activators may be prepared using compounds having formula (A-1) and / or formula (B-1). Specific PK activators and various intermediates prepared by and / or used by the synthetic methods described herein are also disclosed.

The invention relates to a pyruvate kinase-targeted compound shown as a formula I, a preparation method and application thereof, and application of the pyruvate kinase-targeted compound serving as a plant antibacterial agent, and part of the pyruvate kinase-targeted compound shows excellent bactericidal activity.

Described herein are compounds that activate pyruvate kinase, pharmaceutical compositions, and methods of using the same. The compounds are represented by Formula (I), wherein R 1 , R 2 , R a , R b , R j , R k , and Q are as defined herein.

The invention relates to Corynebacterium glutamicum with high indigo yield as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The recombinant corynebacterium glutamicum can be used for overexpression of the following genes: a D-glucose-6-phosphate dehydrogenase gene zwf, a II type deoxyarabinose type heptulose phosphate synthase gene aroG and a flavin-containing monooxygenase gene CcFMO. The recombinant corynebacterium glutamicum can weaken the following genes: a pyruvate kinase gene pyk and a tryptophan synthase gene trpB. The corynebacterium glutamicum is used for overexpressing a zwf gene, an aroG gene and a CcFMO gene, weakening a pyk gene and a trpB gene and enhancing the expression quantity of an indole synthetic pathway related enzyme gene, so that the yield of the corynebacterium glutamicum at a shake flask stage reaches 619mg / L in a fermentation production process of indigo under the condition of not additionally adding tryptophan. The invention provides the recombinant corynebacterium glutamicum with high indigo yield, which is low in production cost, good in stability and high in synthesis efficiency, and is suitable for practical popularization and application.

Provided herein is the use of particular dose levels or amounts of certain pyruvate kinase activators or pharmaceutically acceptable salts or compositions thereof, for treating anemia associated with low risk MDS, lower risk MDS and / or intermediate risk MDS (collectively, LRMDS) and other conditions.

The invention relates to compounds of formula (Ia) and to their use in treating or preventing an inflammatory disease, a disease associated with an undesirable immune response, cancer, obesity, a diabetic disease or a blood disorder:wherein RA, RB, RC and RD, X, Y1, Y2, Y3, Z1, Z2 and m are as defined herein.

To provide a technique for reducing lactate production in cell culture.SOLUTION: The present invention provides a method for reducing or eliminating lactogenic activity in a cell, the method comprising administering to the cell a genetic engineering system, wherein the genetic engineering system knocks down or knocks out expression of a pyruvate kinase muscle (PKM) polypeptide isoform.SELECTED DRAWING: None

The present invention relates to a compound of formula (I) which is 6-((1H-pyrazol-4-yl)sulfonyl)-2-((1-cyclopropyl-1H-pyrazol-3-yl)methyl)phthalazin-1(2H)-one [Formula 1] or a salt and / or solvate thereof; and its use in the treatment or prevention of an inflammatory disease, a disease associated with an unwanted immune response, cancer, obesity, a diabetic disease, or a blood disorder.

The present invention belongs to the technical field of chondroitin sulfate biosynthesis, and discloses a recombinant strain of high-yield chondroitin sulfate and its application and a method for preparing chondroitin sulfate. The recombinant strain provided by the present invention can simultaneously express UDP-glucosamine isomerase, UDP-glucose dehydrogenase, chondroitin polymerase, glucosamine synthetase, phosphoglucosamine mutase, UDP-N-acetylglucosamine pyrophosphorylase, glucose-6-phosphate isomerase, glucose phosphomutase, glucose-1-phosphate uridyl transferase, ATP sulfonase, sulfotransferase, pyruvate kinase and APS kinase, and the above enzyme molecules are combined with the original carbon metabolism pathway in the recombinant strain cell to form an enzyme cascade path with efficient CSA synthesis ability, which can achieve high yield and high purity preparation of CSA, and has the advantages of simple operation, high biological safety of the obtained product and easy large-scale production, and good application prospects.

Provided herein are methods for preparing activators of Pyruvate Kinase (PK) or a salt or a hydrate thereof, as well as intermediates useful in the preparation of such compounds or salts or hydrates thereof. Also provided are a number of crystalline forms of select compounds or salts or hydrates disclosed herein.

The invention discloses a method for detecting the concentration of acetic acid in an anaerobic fermentation system by using a bioelectrochemical sensor, acetic acid is detected by using the bioelectrochemical sensor, the bioelectrochemical sensor comprises a screen-printed electrode, and the working surface of the screen-printed electrode is modified with reduced graphene oxide and polymethylene blue. The working surface of the screen printing electrode is connected with acetokinase, pyruvate kinase and lactic dehydrogenase through a cross-linking agent. According to the method, the adopted sensor has better conductivity, higher response speed and higher current intensity, when the sensor is used for detecting the concentration of acetic acid in a fermentation system, the sensor has the advantages of being easy to operate, low in sample consumption, low in price, high in accurate measurement precision, high in detection speed and the like, and rapid detection of the concentration of acetic acid can be achieved; the acetic acid concentration in the anaerobic fermentation process in different periods can be timely and accurately obtained, effective data can be provided for anaerobic fermentation analysis, and the method has important significance for monitoring the process stability of anaerobic fermentation.

The invention provides an indirect ELISA (Enzyme-Linked Immunosorbent Assay) antibody detection method and an indirect ELISA antibody detection kit based on pyruvate Kinase (PK), and relates to the technical field of parasite detection. The kit comprises a coating antigen Italiella mannici PK protein, a primary antibody, a labeled secondary antibody and an initiator solution, wherein the primary antibody is a mouse or cat anti-Italiella mannici PK protein antibody to be detected; the labeled secondary antibody can be combined with the primary antibody combined with the PK protein of the andrometrium mansoni; and the labeled second antibody is provided with a detection marker. The method comprises the following steps: cloning a PK protein gene into a prokaryotic expression vector PQE80L (+), constructing a recombinant plasmid PQE80L-PK, inducing PK recombinant protein expression by utilizing IPTG (isopropyl-beta-d-thiogalactoside), purifying the recombinant protein to coat an elisa plate, and establishing an indirect ELISA (enzyme-linked immuno sorbent assay) detection method of the Mangnocorynth PK protein antibody by optimizing reaction conditions. The detection method has the advantages of high sensitivity and strong specificity, and provides support for the development of the detection technology of the sitonia mansoni and the prevention and control of the schizocyst.

The present application provides a Pichia pastoris engineering bacterium and a kit and application thereof, a method for obtaining the Pichia pastoris engineering bacterium, a method for improving glucose oxidase yield of the Pichia pastoris, and a method for producing glucose oxidase, wherein the Pichia pastoris engineering bacterium carries a glucose oxidase gene and a pyruvate kinase gene. The Pichia pastoris engineering bacterium of the present application can realize high glucose oxidase yield and high enzyme activity, and lays a good foundation for large-scale production of glucose oxidase.

The application relates to the technical field of engineering bacteria, and particularly discloses recombinant Escherichia coli for synthesizing fucosyllactose as well as a construction method and application thereof. In the genome of the recombinant Escherichia coli, phosphofructokinase genes or phosphofructokinase genes and pyruvate kinase genes are knocked out, so that glucose carbon flow is reduced to flow to a tricarboxylic acid cycle required for cell growth, and the intake of glucose carbon flow is forced to flow more to the synthesis of products (GDP-L-fucose in the synthesis of FL), and the synthesis efficiency of the products is improved. Meanwhile, the glucose carbon flow flowing to the cell growth is greatly reduced, and the glucose effect can be removed, so that the tolerance of the fermentation process to excessive glucose residues is improved, and the difficulty of downstream production amplification is reduced. The recombinant Escherichia coli uses glycerol and glucose compound carbon sources as raw materials, uses lactose as a substrate, and has a 3-fucosyllactose yield of 73.88 g / L in a 5L tank fermentation, which is the highest level reported at present.

Provided herein are methods of treating sickle cell disease, methods of increasing hemoglobin levels in a subject diagnosed with sickle cell disease, and methods of reducing hemolysis-associated complications in a subject diagnosed with sickle cell disease, together with kits for such treatments, in a subject in need thereof, comprising administering a Gardos channel inhibitor such as 2,2-bis(4-fluorophenyl)-2-phenylacetamide and a pyruvate kinase activator such as N-(4-((4-(Cyclopropylmethyl)-1-piperazinyl)carbonyl)phenyl)-8- quinolinesulfonamide to the subject. Subjects eligible for treatment include subjects as having hemolysis dominant (HD) sickle cell disease on the basis of laboratory markers, clinical history, and / or sequelae.

The present invention relates to a use of pyruvate kinase. Specifically, the pyruvate kinase of the present invention can be used to prepare a composition for (a) preventing and / or treating skin pigmentation; (b) whitening skin tone; and / or (c) delaying melanoma progression. The pyruvate kinase of the present invention can significantly inhibit melanin synthesis, including melanoma cell synthesis, and has excellent skin whitening effects and reduces persistent pigment deposition in melanoma cells.

The invention discloses recombinant escherichia coli, a construction method thereof and application of the recombinant escherichia coli in production of 3-dehydroshikimic acid, and belongs to the technical field of bioengineering. According to the recombinant escherichia coli, escherichia coli FMME-DHS03 is used as an original strain, pyruvate kinase gene pykA is knocked out, excessive 3-dehydroquinic acid synthase gene aroB, transketolase gene tktA and galactose permease gene galP are added, the finally obtained recombinant strain is fermented for 40 h, the yield of 3-dehydroshikimic acid reaches 130.9 g / L, and the yield of glucose reaches 0.55 g / g. The process for producing the 3-dehydroshikimic acid through fermentation is simple and easy to operate, free of additional antibiotics, short in period, high in yield, low in culture medium cost and suitable for industrial production.