56 results about "Alpha-oxoglutarate" patented technology

The invention discloses a method for efficiently producing alpha-oxoglutarate by adopting whole-cell transformation. L-amino acid deaminase genes transformed by easily wrong PCR (Polymerase Chain Reaction) or site-saturation mutagenesis are expressed through bacillus subtilis, the transformation rate of the alpha-oxoglutarate produced by whole-cell transformation of L-glutamic acid is increased, and the gene sequence of the L-amino acid deaminase genes is shown as SEQ ID NO.1 or SEQ ID NO.2. A whole-cell transformation system in which the transformation rate is increased is constructed, the problem that a chemical method for synthesizing alpha-oxoglutarate is tedious in steps, low in yield and causes environmental pollution and the problem that the transformation rate during production of the alpha-oxoglutarate is low through enzymatic transformation are solved, the alpha-oxoglutarate is produced in a pollution-free mode at high yield through a one-step method, and a certain theoretical basis is laid for subsequent industrial production.

The invention discloses a pig feed additive for improving intestinal health and boosting immunity and belongs to the field of feed additives. The additive is prepared from compound plant polysaccharide, complex enzyme preparation, lactobacillus plantarum powder, lactobacillus buchneri powder, stevia rebaudiana powder, fructus chebulae powder, orange peel powder, arginine, glutamine dipeptide, alpha-oxoglutarate, cystine, taurine and a carrier, wherein corncob powder and peanut shell powder serve as the carrier. By the adoption of the additive, pig intestinal health can be improved, intestine development of live pigs can be promoted, the feed digestibility and absorptivity of live pigs can be improved, pig growth can be promoted, marketing period can be shortened, economical benefits can be increased, and feeding effect can be improved. Meanwhile, the immunity of pigs can be improved, the morbidity of pigs can be reduced, and application prospects are quite broad.

The invention relates to a preparation method of an alpha-oxoglutarate fluorescent / ultraviolet molecular probe and application of the alpha-oxoglutarate fluorescent / ultraviolet molecular probe to biological samples. The preparation method includes the steps that NBD-Cl is dissolved in chloroform, the dissolution concentration is 0002-0.012 g / ml, then a hydrazine hydrate-methanol solution with the volume concentration being 0.2-1.2% is added in and uniformly mixed, brown precipitate is obtained through stirring at room temperature and filtered, a filter cake is washed through ethyl acetate and dried, and the brown product alpha-oxoglutarate fluorescent / ultraviolet molecular probe is obtained. The alpha-oxoglutarate fluorescent / ultraviolet molecular probe is applicable to qualitatively and quantitatively analyzing alpha-oxoglutarate in the biological samples, and detection is sensitive, accurate and fast. The biological samples mainly comprise serum, living cells, muscle tissue and the like. The alpha-oxoglutarate fluorescent / ultraviolet molecular probe can be applied to analytical chemistry, life organic analysis, disease pre-diagnosis, medical clinical inspection and other related fields. The alpha-oxoglutarate fluorescent / ultraviolet molecular probe has the advantages of being good in response performance and high in data accuracy, repeatability and precision, equipment is convenient, fast and easy to operate, operability is high, and the alpha-oxoglutarate fluorescent / ultraviolet molecular probe is particularly suitable for large-batch sample combined screening and other large-data research.

The invention discloses a method for increasing yield of alpha-oxoglutarate produced through whole-cell transformation, and belongs to the technical field of biotechnology. The method is characterized in that recombinant bacillus subtilis is used as a producing bacterial strain, and an sucA gene in a genome of the recombinant bacillus subtilis is knocked out. The sucA gene is knocked out so as to block the decomposition approach of intracellular L-glutamic acid, so that the bacterial strain is transformed so as to efficiently transform L-glutamic acid for producing alpha-oxoglutarate, thus the problems of high cost, low yield and serious pollution of industrialized alpha-oxoglutarate are solved.

The invention relates a rhizobium japonicum culture medium and a method for preparing a liquid rhizobium japonicum agent by adopting the rhizobium japonicum culture medium and aims at solving the problem that an existing liquid rhizobium japonicum agent is unbalanced in nutrition and low in thallus activity and rhizobium japonicum prepared by means of a fermentation technology cannot stably survive at the room temperature. The rhizobium japonicum culture medium is an aqueous solution prepared by glycerin, mannitol, glucose, rhamnose, fructose, alpha-oxoglutarate, yeast powder, serine, arginine, NH4C1, K2HPO4, KH2PO4, MgSO4*7H2O, CaCl2*2H2O, FeSO4*7H2O, NaCl, Na2MoO4*2H2O and biotin, and can be obtained by controlling fermentation tank conditions. The viable count of microbial inoculum of the rhizobium japonicum culture medium preserved at 20 DEG C for 12 months can still reach to more than 100 hundred millions per milliliter.

The invention relates to an application of Chinese yarn glycoprotein in improvement of the activities of an oxidative metabolism enzymes of mitochondria of a brain cell. Selection and verification are carried out by an animal test so as to prove that the Chinese yarn glycoprotein can obviously improve the activities of key enzymes namely pyruvate dehydrogenase and alpha-oxoglutarate dehydrogenase used for oxidative metabolism in the mitochondria of the brain cell, so that brain cell deterioration and damage diseases, such as senile dementia, vascular dementia, cerebral infarction and cerebral palsy, accompanying with oxidative metabolism efficiency reduction caused by reduction of the activities of the enzymes can be effectively treated.

The invention relates to a method for producing L-4-hydroxyisoleucine by a microbial enzyme conversion method. The method comprises the following steps: 1, culturing a mature bacterial strain; 2, grafting mature escherichia coli recombinant bacteria which are capable of inducing production of L-4-hydroxyisoleucine into a fermentation tank, and carrying out fermentation culture, wherein the temperature of the fermentation process is controlled in two stages, i.e., 4 to 36 DEG C in the earlier stage and 30 to 32 DEG C in the later stage; 3, collecting thallus after fermentation, adding phosphate buffer and washing, then adding alpha-oxoglutarate, isoleucine, ferrous sulfate, magnesium sulfate and ascorbic acid, and carrying out forced ventilation for catalysis; 4, removing bacteria from the catalytic liquid with a ceramic membrane, filtering with an ultrafiltration membrane, decoloring with activated carbon, desalting with faintly acid cation exchange resin, adsorbing effluent with 732 cation exchange resin and eluting with ammonia water, carrying out evaporation concentration and recrystallization to obtain the L-4-hydroxyisoleucine. The preparation method disclosed by the invention is simple and efficient in reaction system and simple and feasible in extraction process. The conversion rate is up to over 95.4 percent, and the purity of the product can reach over 98 percent.

A method with adjuvant factor to regulate the carbon metabolism to accomplish the overdose accumulation of alpha-ketoglutarate (alpha-KG) is provided, which belongs to the technical field of fermentation process optimized by the strategy of adjuvant factor regulation. The invention adopts the method of adjuvant factors including vitamins and metal ions, etc. which are added into the fermentation process with the sub-proper amount, and then the pathway flux of PDH and PC are increased selectively to accomplish the overdose accumulation of alpha-KG. During the process of the production of alpha-KG through fermentation of T.glabrata, the concentration of vitamin B1 is raised to increase the accumulated alpha-KG to 10.3g / L; then the biotin concentration is raised to increase the accumulated alpha-KG to 43.7g / L, and the concentration of pyruvic acid is decreased to 21.8g / L. The strategy also has the significance to guide the production of the other fermented products.

The invention discloses application of alpha-oxoglutarate to improving the heat production capability of fat animals and reducing the body fat content. The invention provides new application of alpha-oxoglutarate, alpha-oxoglutarate can obviously improve whole heat production of high fat diet animal bodies and heat production of brown fat, can prominently improve expression of brown fat heat producing marker genes UCP1, PRDM16, PGC1alpha, Cidea and Elovl3, and meanwhile, can reduce the overall body fat content of the high fat diet animal bodies and the fat weight of different parts. In addition, alpha-oxoglutarate does not contain any substance harmful to animals or human body, the adding amount is less, and alpha-oxoglutarate can be directly added into animal drinking water, fodder, foodor drugs, has the advantages of safety, effectiveness and no toxic and side effects, has the wide application prospect at the aspect of improving heat production of the high fat diet animals and reducing the body fat content, and explores a new means for treating obesity and related diseases.

The invention discloses a blocking agent for reducing the content of heavy metal of edible parts of crops and an application method thereof. The blocking agent comprises glycerol or one or two of glycerol, oxaloacetic acid and alpha-oxoglutarate, and can realize the purposes of blocking absorption of heavy metal and transferring to edible parts through foliage spraying after vegetable farming andbefore harvesting. The blocking agent can obviously reduce the contents of heavy metal, such as plumbum and cadmium, at the edible parts of vegetables, so as to ensure that vegetables produced in mildor medium-heavy metal contaminated vegetable fields meet food safety standards. The blocking agent is low in effective spraying concentration, can be mixed with pesticide for controlling vegetable diseases and insect pests and then be sprayed, and is low in cost.

The invention provides an enzyme activity detection method of Fe and alpha-oxoglutarate dependence dioxygenases, comprising a step of detecting the enzyme activity of the enzyme by a color reaction of inorganic phosphate generated by a coupled reaction of Fe and oxoglutarate dependence dioxygenases and succinyl-coenzyme A synthetase and ammonium molybdate malachite green dye. In addition, the invention also provides an enzyme activity detection system of dioxygenase, a kit and application of thereof.