59 results about "Glutamate synthetase" patented technology

The invention discloses a high-efficiency production method of gamma-polyglutamic acid, and belongs to the technical field of production of polymer materials based on biological fermentation. According to the method, bacillus licheniformis CGMCC (China General Microbiological Culture Collection Center) NO.3336 for high-efficient production of gamma-polyglutamic acid is taken as a starting strain, sodium nitrate is added into a fermentation culture medium, and the activity of polyglutamate synthase is influenced under the nitrate respiration condition by adjusting and controlling the pH value and the ventilation quantity in a fermentation process, so that residual glutamic acid in a fermentation solution is reduced and the conversion rate of glutamic acid is increased; after fermentation, the glutamic acid content of the fermentation solution is 5-15 g / L and the gamma-polyglutamic acid yield is 45-55 g / L. Therefore, the economic and feasible production method is provided for industrialization production of gamma-polyglutamic acid.

The invention discloses a production method of a gamma-polyglutamic acid with high molecular weight, and belongs to the technical field of biological fermentation production of polymer materials. Bacillus licheniformis CGMCC NO.3336 is taken as an original strain, the rotating speed is reduced at a later stage of fermentation, the effect on polyglutamic acid synthesis caused by shearing force is reduced by regulating and controlling of ventilation and rotation speed in the fermentation process, and the temperature is changed, so as to affect the activity of folylpolyglutamate synthetase. Especially, the fermentation pH is controlled and the ion strength in the culture medium is increased by feeding glucose, sodium chloride, CaCl2.6H2O, MnSO4 and MgSO4, so as to obtain the polyglutamic acid with accurate molecular weight. The molecular weight is high, and the molecular weight of the gamma-polyglutamic acid is 1000000-4000000 Dalton by detection after fermentation is finished. The method is simple in technology, and the gamma-polyglutamic acid is high in molecular weight and relatively stable and controllable, and large-scale production can be achieved.

Sensitivity of a patient's cancer to treatment with 10-propargyl-10-deazaaminopterin is assessed and patients are selected for treatment of cancer with 10-propargyl-10-deazaaminopterin, by determining the amount of a selected polypeptide expressed by the cancer and comparing the amount with the amount of the selected polypeptide expressed by a reference cancer. The polypeptide includes a member of a folate pathway polypeptide within a cell and may include at least one of reduced folate carrier-1 enzyme (RFC-1), dihydrofolate reductase (DHFR), folylpoly-gamma-glutamate synthetase (FPGS), thymidylate synthase (TS), γ-glutamyl hydrolase (GGH), and glycinamide ribonucleotide formyltransferase (GARFT).

Expression vectors are described that can efficiently produce virion capsid protein, tumor-associated protein of human papillomavirus on a microbial surface. Bacterial strains harboring such surface display vectors, and the use of the bacterial strains or their extracts or purified products as complex vaccines, are also described. The surface display vectors contain one or more than two genes selected from among pgsB, pgsC and pgsA, encoding a poly-χ-glutamic acid synthetase complex (pgsBCA) of a Bacillus sp. strain, and genes that encode virion capsid proteins, tumor-associated proteins of human papillomavirus. Methodology for preparing the foregoing vectors, vaccines and transformed microorganisms are also described.

The invention relates to a special plant expression vector pH2-35S-PrbcS-GS1 which comprises an arabidopsis thaliana cytoplasm glutamine synthetase gene GS1 and can improve the utilization rate of a plant nitrogen element. A method of RT-PCR is used for cloning the GS1 gene from the arabidopsis thaliana of a model plant, a photoinduction type promotor (the promotor of a a small subunit Rubisco) is used for controlling the excessive expression of the GS1 gene in a plant leaf and a leaf disc conversion method is used for transferring the GS1 gene into a pPZP221-PrbcS-Dof1 type transgene tobacco. An experiment result shows that the GS1 gene can be normally transferred in the transgene tobacco; under the nutrition condition of low nitrogen and the growing conditions of indoor irradiation for 24 hours of 2000LUX and 25 DEG C, the growing situation of the plant transferred with the single gene of Dof1 is (the expression of the gene is controlled by the photoinduction type promotor Prbcs) is a little better than that of a contrast tobacco (a wild type without transgene); after being transferred under the natural growing condition of a green house, the growing situation of the tobacco which is simultaneously transferred with the GS1 gene and the Dof1 gene shows remarkable growing advantages than that of the contrast plant; and therefore, simultaneously and excessively expressing the GS1 gene and the Dof1 gene, can improve the efficiency of the GS / GOGAT (glutamine synthetase / glutamic acid synthetase) approaches in the leaf more extensively, thereby improving the utilization rate of the plant nitrogen element. The vector can be broadly applied to the molecule breeding of crops, improving the utilization rate of the plant nitrogen element thereof and the durability to the nutrition condition of low nitrogen and being capable of obtaining a higher yield under the conditions of applying less fertilizers and even not applying the fertilizers.

The invention discloses application of gamma-aminobutyric acid in improving salt stress resistant capacity of corn, relates to application of gamma-aminobutyric acid in growth and development of the corn, and aims at providing new application of gamma-aminobutyric acid. Gamma-aminobutyric acid has a regulating effect on oxidative damage and enzymatic activity of corn leaves under salt stress, the content of malonaldehyde in corn seedling leaves under the salt stress is decreased, the content of superoxide anions in the leaves is decreased, the content of superoxide dismutase in the leaves is decreased, improvement of peroxidase activity is promoted, the activity of an antioxidase system under NaCl stress is improved, the regulating effect on corn seedling nitrogen metabolism under the salt stress is achieved, the content of soluble protein of the corn leaves and root systems under the salt stress is increased, and pyruvic acid content, glutamine synthetase activity, glutamate synthase activity, glutamate dehydrogenase activity and glutamate decarboxylase activity in the corn leaves and the system roots under the salt stress are decreased. The application of gamma-aminobutyric acid in improving the salt stress resistant capacity of the corn is used in the field of corn cultivation.

The present invention relates to a surface expression vector of SARS coronavirus antigen containing a gene encoding an antigen of SARS inducing coronavirus and any one or two or more of genes pgsB, pgsC and pgsA encoding poly-gamma-glutamic acid synthase complex, a microorganism transformed by the surface expression vector, and a SARS vaccine comprising the microorganism. According to the present invention, it is possible to economically produce a vaccine for prevention and treatment of SARS using a recombinant strain expressing an SARS coronavirus antigen on their surface.

Expression vectors are described that can efficiently produce virion capsid protein, tumor-associated protein of human papillomavirus on a microbial surface. Bacterial strains harboring such surface display vectors, and the use of the bacterial strains or their extracts or purified products as complex vaccines, are also described. The surface display vectors contain one or more than two genes selected from among pgsB, pgsC and pgsA, encoding a poly-χ-glutamic acid synthetase complex (pgsBCA) of a Bacillus sp. strain, and genes that encode virion capsid proteins, tumor-associated proteins of human papillomavirus, Methodology for preparing the foregoing vectors, vaccines and transformed microorganisms are also described.

The present invention relates to a surface expression vector having pgsBCA, a gene coding poly-gamma-glutamate synthetase and a method for expression of target protein at the surface of microorganism using the vector. The vector, in which foreign genes are inserted, transforms microorganisms and makes foreign proteins expressed stably on the surface of microorganisms.

The invention discloses a special plant expression vector pK2-35S-Prbcs-*T-GS2 which contains an arabidopsis chloroplast type glutamine synthetase gene (GS2) and can improve the metabolic ability of plant nitrogen. A RT-PCR method is utilized to clone a GS2 gene from a model plant Arabidopsis, a promoter of a Rubisco small subunit with the light-inducible promoter is used to control the overexpression of the GS2 gene in plant leaves, and the GS2 gene is transformed into wild tobacco through a leaf disk transformation method. The experimental result shows that the GS2 gene can perform normal transcription in transgenic tobacco and the growth condition of a strain with the GS2 gene is better than that of a check plant (un-transformed wild type) under low nitrogen nutrition condition, which indicates that the overexpression of the GS2 gene can improve the efficiency of a GS / GOGAT (glutamine synthetase / glutamate synthetase) way to a great extent, thereby improving the capability of the assimilation of the plant nitrogen. The special vector has the advantages that the special vector can be used for molecular breeding of crops, improves the utilization rate of nitrogen and the tolerance to low nitrogen stress, and obtains higher yield under the condition of small nitrogenous fertilizer application or even no nitrogenous fertilizer application.

The invention relates to the field of peanut cultivation techniques and in particular relates to a special growth regulator for peanuts in a fruiting period. The special growth regulator comprises 20-30g of paclobutrazol wettable powder, 2.5-3.0g of chitosan oligosaccharide, 50-100g of seaweed extract and 115-180g of saccharose. The content of chlorophyll of leaves, the leaf net photosynthetic rate and the root activity of the peanuts treated by the regulator in the fruiting period are obviously higher than corresponding contrasts, the leaf SOD (superoxide dismutase) activity and the POD (peroxidase) activity are both obviously higher than corresponding contrasts, the content of malondialdehyde is obviously lower than the contrast, and therefore, the regulator can be used for well eliminate oxygen free radicals and delayin g senescence. By utilizing the special growth regulator, the leaf nitrate reductase activity, the glutamine synthetase and glutamate synthase activities are obviously improved, the leaf sucrose synthetase and sucrose phosphate synthase activities are improved, and the accumulation of pod dry matters and compounds of nucleus protein and fat are facilitated. The peanut pod yield is obviously increased, the production increase reason is that the weight of single fruits is obviously increased, the full fruit rate is increased, and the double-kernel fruit rate and the kernel yield rate are greatly increased simultaneously.

The present invention relates to a surface expression vector having pgsBCA, a gene coding poly-gamma-glutamate synthetase and a method for expression of target protein at the surface of microorganism using the vector. The vector, in which foreign genes are inserted, transforms microorganisms and makes foreign proteins expressed stably on the surface of microorganisms.

The present invention relates to a method for assessing the sensitivity of a patient's cancer to treatment with 10-propargyl-10-deazaaminopterin and a method for selecting a patient for treatment of cancer with 10-propargyl-10-deazaaminopterin, by determining the amount of a selected polypeptide expressed by the cancer and comparing the amount with the amount of the selected polypeptide expressed by a reference cancer, wherein the polypeptide includes a member of folate pathways within cells and may include at least one of reduced folate carrier-1 enzyme (RFC-1), dihydrofolate reductase (DHFR), folylpoly-gamma-glutamate synthetase (FPGS), thymidylate synthase (TS), γ-glutamyl hydrolase (GGH), and glycinamide ribonucleotide formyltransferase (GARFT). The present invention also relates to the use of 10-propargyl-10-deazaaminopterin in the treatment of multiple myeloma.

The present invention relates to a method for expressing each of peptide antibiotics P5 3 and Ana13 35 having amphiphilicity and showing antibacterial, antifungal and anticancer activities 61, 63, 65, 67, 69, 71, on the microbial surface, using a vector containing outer membrane protein genes (pgsBCA) that are derived from Bacillus sp. strains and involved in the synthesis of poly-gamma-glutamate. Moreover, the present invention relates to lactic acid-forming bacteria having each of the peptide antibiotics P5 15 and Ana13 43 expressed on their surface, and the use thereof.According to the present invention, the peptide antibiotics can be expressed on the surface of various microorganisms transformed with the surface expression vectors. The inventive method for the surface expression of the peptide antibiotics allows the peptide antibiotics to be mass-produced without a purification process. Thus, the inventive method has very high industrial applicability. Further, the present invention can be applied to other peptide antibiotics besides P5 3 and Ana13 35.

The present invention relates to a surface expression vector for HPV vaccine preparation including a gene that encodes an antigen protein related to the occurrence of human papilloma virus tumors, connected with: a composite gene with repE mutation gene having the amino sequence of sequence No. 1, a promoter, a poly-gamma-glutamate synthetase complex, and a poly-gamma-glutamate synthetase complex gene. The recombinant lactobacillus of the present invention that has been transformed into a surface expression vector of the antigen protein of the human papilloma virus, and the composition using the same, may be utilized as a treatment vaccine for cervical cancer and directly applied to the cervical or vaginal area, so that it has very economical effects.

The invention is about a method of measuring the content of blood ammonia, and it also concerns the reagent box of blood ammonia diagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, glutamate, 2ú¡ketoglutarate, reduced coenzyme, adenosine triphosphate, glutamine synthetase, glutamate synthetase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get the content of blood ammonia. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

The present invention relates to a surface expression vector of SARS coronavirus antigen containing a gene encoding an antigen of SARS inducing coronavirus and any one or two or more of genes pgsB, pgsC and pgsA encoding poly-gamma-glutamic acid synthase complex, a microorganism transformed by the surface expression vector, and a SARS vaccine comprising the microorganism. According to the present invention, it is possible to economically produce a vaccine for prevention and treatment of SARS using a recombinant strain expressing an SARS coronavirus antigen on their surface.

The invention provides a growth regulator for the fruiting period of peanuts. The growth regulator comprises the following components in percentage: 30-35% of growth promoter and 65-70% of growth nutrient solution. The regulator can favorably remove oxygen free radicals and delay senility; and the regulator obviously improves the activity of leaf nitrate reductase and the activities of glutamine synthetase and glutamate synthetase, and improves the activities of leaf sucrose synthetase and sucrose phosphate synthetase, thereby being beneficial to the accumulation of dry matters in pods and the synthesis of protein and fat in seed kernels. The growth regulator obviously increases the yield of peanut pods; and the yield is increased because the weight of a single pod is obviously increased, the full-pod rate is improved, and the dual-kernel pod rate and the kernel rate are greatly improved.

The present invention relates to a surface expression vector expressing a parvovirus antigen on the surface of microorganisms, the vector containing not only a gene encoding the capsid antigen protein of parvovirus causing canine parvovirus (CPV) infection and feline panleukopenia (FLP) but also at least one of pgsB, pgsC and pgsA genes encoding a poly-gamma-glutamate synthetase complex. Also, the present invention relates to microorganisms transformed with the surface expression vector, and parvovirus vaccines containing the transformed microorganisms. According to the present invention, the use of the recombinant bacterial strains expressing the parvovirus antigen on their surface allows the economical production of vaccines for the treatment and prevention of canine parvovirus infection and feline panleukopenia.

The invention discloses a production method of a gamma-polyglutamic acid with high molecular weight, and belongs to the technical field of biological fermentation production of polymer materials. Bacillus licheniformis CGMCC NO.3336 is taken as an original strain, the rotating speed is reduced at a later stage of fermentation, the effect on polyglutamic acid synthesis caused by shearing force is reduced by regulating and controlling of ventilation and rotation speed in the fermentation process, and the temperature is changed, so as to affect the activity of folylpolyglutamate synthetase. Especially, the fermentation pH is controlled and the ion strength in the culture medium is increased by feeding glucose, sodium chloride, CaCl2.6H2O, MnSO4 and MgSO4, so as to obtain the polyglutamic acid with accurate molecular weight. The molecular weight is high, and the molecular weight of the gamma-polyglutamic acid is 1000000-4000000 Dalton by detection after fermentation is finished. The method is simple in technology, and the gamma-polyglutamic acid is high in molecular weight and relatively stable and controllable, and large-scale production can be achieved.

The invention discloses a high-efficiency production method of gamma-polyglutamic acid, and belongs to the technical field of production of polymer materials based on biological fermentation. According to the method, bacillus licheniformis CGMCC (China General Microbiological Culture Collection Center) NO.3336 for high-efficient production of gamma-polyglutamic acid is taken as a starting strain, sodium nitrate is added into a fermentation culture medium, and the activity of polyglutamate synthase is influenced under the nitrate respiration condition by adjusting and controlling the pH value and the ventilation quantity in a fermentation process, so that residual glutamic acid in a fermentation solution is reduced and the conversion rate of glutamic acid is increased; after fermentation, the glutamic acid content of the fermentation solution is 5-15 g / L and the gamma-polyglutamic acid yield is 45-55 g / L. Therefore, the economic and feasible production method is provided for industrialization production of gamma-polyglutamic acid.

The invention relates to a reagent (kit) for diagnosing / determining amino acid by utilizing the techniques of an enzymatic recycling method, an enzyme colorimetric method and an enzyme coupling method, and also relates to a method for determining the concentration of amino acid and a composition and components of the reagent, belonging to the technical field of detection and determination of medicine / foodstuff / environment. The reagent (kit) mainly comprises the following components: a buffer solution, coenzyme, hydrogen peroxide, 2-ketoglutarate, amino acid dehydrogenase, glutamate oxidase, glutamate synthetase and a stabilizer. A sample and the reagent are mixed according to a certain volume ratio to generate a series of enzymic reactions, then a reactant is placed under an ultraviolet / visible light analyzer, and the rising speed of absorbance in the position of 340 nm of main wave length is detected so that the concentration of the amino acid is measured and calculated.

The invention adopts a double enzyme coupling system of a G-quadruplet-Hemin peroxide mimic enzyme and a glutamine synthetase; the selectivity of the glutamine synthetase is used for specifically recognizing glutamine; the glutamine is quantitatively converted into a fluorescence signal; the content of glutamine in a sample is detected via a standard curve method. The detection method is convenient to operate, low in cost, quick, accurate, high in sensitivity, and relatively small in damage to the sample, and can be widely applied to the field of biology and agriculture.

The present invention relates to the determination method of the content of glycine, the composition and the component of the reagent using double amplification method, enzyme colorimetric method and enzyme-linked method technology, and the technical principle of its determination is based on hydrogen cyanide synthase, glutamic acid decarboxylase 1. The series of catalyzed reactions of glutamic acid synthetase are completed, and the present invention also relates to a glycine determination kit. The determination method of the invention has high sensitivity and small error, so the determination method and the kit of the invention can be widely used in food inspection.

The invention relates to the technical field of peanut cultivation, in particular to a special growth regulator for peanut pod setting stage: paclobutrazol wettable powder 20-30 g, chitosan oligosaccharide 2.5-3.0 g, seaweed extract 50-100 g, sucrose 115-180 g g. The chlorophyll content, net photosynthetic rate, and root activity of peanut leaves treated with this regulator at the pod stage were significantly higher than those of the controls, and the SOD activity and POD activity of the leaves were significantly higher than that of the controls, while the malondialdehyde content was significantly lower than that of the controls, indicating that Using this regulator can remove oxygen free radicals very well and delay aging; this regulator can significantly improve the activity of leaf nitrate reductase, glutamine synthetase and glutamic acid synthase, and increase the activity of leaf sucrose synthase and sucrose synthase. Phosphate synthase activity is beneficial to the accumulation of dry matter in pods and the synthesis of protein and fat in seeds. Significantly increased the yield of peanut pods. The reason for the increased yield is that the weight of a single fruit was significantly increased, the rate of full fruit was increased, and the rate of double-seed fruit and the rate of kernels were greatly increased.

The invention relates to a homocysteine diagnostic kit utilizing technologies of an enzymatic-colorimetric method and an enzyme-link method, also relates to a method and a principle of measuring homocysteine concentration and compositions and components of reagents, and belongs to the technical field of testing and measuring of medical science. The kit mainly comprises the following compositions: buffer solution, reduced coenzyme, glutamic acid, adenosine triphosphate, 2-oxoglutaric acid, homocysteine-alpha, gamma-lyase, glutamine synthetase, glutamate synthase, and stabilizer; samples are mixed with the reagents in certain volume ratio to perform a series of enzymatic reactions; then reactants are placed under a UV / visible analyzer; and the descending speed of absorbance is tested at the position where dominant wave length is 340nm so as to measure and calculate the homocysteine concentration.

A surface expression vector for preparing HPV vaccines, in which the surface expression vector contains a gene encoding a repE mutant protein having an amino acid sequence of SEQ ID NO: 1, a promoter, a poly-gamma-glutamate synthetase complex gene, and a gene which is linked with the poly-gamma-glutamate synthetase complex gene and encodes a tumor induction-associated antigen protein of human papillomavirus.An expression vector constitutively expressing a high level of the human papillomavirus (HPV) antigen protein is provided. Also, a recombinant lactic acid bacteria, transformed with the expression vector and expressing the HPV antigen protein on the surface thereof, and a composition comprising the recombinant lactic acid bacteria are provided. The recombinant lactic acid bacteria and the composition are very effective as a vaccine for the treatment of cervical cancer, because they can be applied orally or directly to the vagina.

The present invention relates to a method for assessing the sensitivity of a patient's cancer to treatment with 10-propargyl-10-deazaminopterin and a method for selecting a patient for treatment of cancer with 10-propargyl-10-deazaminopterin, by determining the amount of a selected polypeptide expressed by the cancer and comparing the amount with the amount of the selected polypeptide expressed by a reference cancer, wherein the polypeptide includes a member of folate pathways within cells and may include at least one of reduced folate carrier-1 enzyme (RFC-1), dihydrofolate reductase (DHFR), folylpoly-gamma-glutamate synthetase (FPGS), thymidylate synthase (TS), γ-glutamyl hydrolase (GGH), and glycinamide ribonucleotide formyltransferase (GARFT). The present invention also relates to the use of 10-propargyl-10-deazaminopterin in the treatment of multiple myeloma.

The invention relates to a reagent (kit) for diagnosing / determining amino acid by using an enzyme colorimetric method and an enzyme link method, and also discloses a method for determining the concentration of the amino acid, a composition and components of the reagent, belonging to the technical field of medicine / food / environmental test determination. The reagent (kit) comprises the main components: a buffer solution, a coenzyme, 2-ketoglutarate, an amino acid oxidase, a glutamate oxidase, a glutamate synthetase and a stabilizing agent. A sample and the reagent are mixed according to a certain volume ratio to generate a series of enzymatic reactions, and reactants are placed under an ultraviolet / visible light analyzer for detecting the degree of absorbance rise at the dominant wavelength of 340 nm, thereby measuring and calculating the concentration of the amino acid.

The present invention relates to the measuring method of the content of glycine, the composition and the component of the reagent by using enzyme cycle amplification method, enzyme colorimetric method and enzyme-linked method technology, the technical principle of its measuring is based on glycine transaminase, glutamine synthetase, glutamine A series of catalyzed reactions of the acid synthase is completed, and the invention also relates to a glycine assay kit. The determination method of the invention has high sensitivity and small error, so the determination method and the kit of the invention can be widely used in food inspection.