154 results about "Glucose phosphate" patented technology

The invention provides a group of relevant enzymes for preparing mannitol by performing anabolism on Chinese caterpillar fungus serving as a multifunctional production fungus and hirsutella sinensis based on glucose, a gene for encoding these enzymes and application thereof. The relevant enzymes include (1) hexokinase: manA1-A6 proteins of which the sequences are SEQ ID No.1-6, (2) phosphoglucoisomerase: manB1-B3 proteins of which the sequences are SEQ ID No.7-9, and (3) mannitol-1-P dehydrogenase: manC protein of which the sequence is SEQ ID No.10. In the invention, detailed researches are performed on the metabolic pathway of mannitol synthesized by using Chinese caterpillar fungus serving as a multifunctional production fungus, hirsutella sinensis and glucose on the aspect of principle, cloned DNA (Deoxyribose Nucleic Acid) comprising a nucleotide sequence provided by the invention can be transferred into engineering bacteria with transduction, conversion and conjugal transfer methods, and host mannitol is endowed with high expression by regulating the expression of a biosynthetic gene of the mannitol.

Provided is a recombinant strain producing L-amino acids, a constructing method therefor and a method for producing L-amino acids. Compared with the starting strains, the recombinant strain producing L-amino acids has decreased expression of 6-phosphoglucose isomerase Pgi and increased expression of 6-phosphogluconate dehydrogenase Zwf-OpcA. The starting strain can accumulate objective amino acids. Fermentation culturing of the recombinant strain can increase the yield of L-amino acids significantly. Also provided is a novel method for increasing the fermentation yield of L-amino acids, capable of being be used in bacterial fermentation of L-amino acids.

The invention discloses a freeze-drying concentrated glucose detection reagent microsphere and a preparation method thereof. The freeze-drying microsphere comprises a triethanolamine buffer solution, ATP, NADP, hexokinase, 6-glucose dehydrogenase phosphate, a preservative, trehalose, polyethylene glycol 8000, polyethylene glycol 20000, TrionX-100 and the like. The preparation method comprises the following steps: (1) preparing a solution, and performing filtration and degassing; (2) preparing droplets by using a precise quantification liquid separation system, dropwise adding the droplets into liquid nitrogen, and preparing a freezing microsphere; and (3) transferring the freezing microsphere into a freeze dryer, and performing freeze-drying on the freezing microsphere to obtain a freeze-drying detection reagent microsphere. A glucose detection reagent provided by the invention is a spherical granular freeze-drying reagent and can be pre-packaged into a detection chip. Compared with an existing liquid detection reagent, the glucose detection reagent has the advantages that the storage, transportation and the like at the stable room temperature can be realized. Compared with a freeze-drying powder reagent, the glucose detection reagent has the advantages of accuracy in quantification, convenience in packaging treatment and the like.

The invention relates to a high-yield acetyl-glucosamine recombinant bacillus subtilis and a construction method thereof. Bacillus subtilis BSGN6 serves as an original strain, glucokinase encoding gene glck expression and glucose phosphate isomerase encoding gene pgi expression in the bacillus subtilis are regulated by xylose inducible promoters PxylA, moderate expression of glck and pgi genes is controlled, the yield of acetyl-glucosamine in the recombinant bacillus subtilis is greatly improved, and a foundation is laid for further producing glucosamine by modifying the bacillus subtilis in metabolic engineering. The construction method is simple and easy to operate, the yield of acetyl-glucosamine can be greatly improved by the constructed recombinant bacillus subtilis, and the recombinant bacillus subtilis has a good application prospect.

The invention belongs to the technical field of gene engineering, and particularly relates to an escherichia coli gene engineering bacterium and a method for producing L-threonine by using the escherichia coli gene engineering bacterium. The gene engineering bacterium is characterized in that a promoter P<ppc> of phosphoenolpyruvate carboxylase gene (ppc) of an initial strain is replaced into a promoter P<zwf> of 6-phosphate dehydrogenase gene (zwf), so that the goal of regulating the L-threonine production capacity by glycine betaine is achieved. In the fermentation process, the glycine betaine is added, so that the L-threonine yield through shaking flask fermentation can reach 50 to 55g / L; the 5L fermentation tank yield reaches 120 to 150g / L; the saccharic acid conversion rate reaches 59 to 61 percent.

The invention provides a chassis system for ATP (Adenosine Triphosphate) regeneration and application. The chassis system for ATP regeneration comprises the following five enzymes: alpha-glucan phosphorylase, glucophosphomutase, phosphoglucose Isomerase, phosphoketolase and acetokinase. In addition, the chassis system can further comprise the following seven enzymes: transaldolase, transketolase,ribose-5-phosphate isomerase, ribulose-3 epimerase, triosephosphate isomerase, fructose-bisphosphate aldolase and fructose-1,6-bisphosphatase. The chassis system can be coupled to an enzymic catalyticreaction needing ATP, utilizes starch or maltodextrin as energy, is not added with any coenzyme, can efficiently regenerate the ATP at low cost through a one-pot reaction and is an economical, highlyefficient, stable and sustainable ATP regeneration system.

The invention discloses a serum-free immune cell culture medium. The serum-free immune cell culture medium is prepared mainly from amino acids, vitamins, minerals, D-glucose, 4-hydroxyethyl piperazinyl ethanesulfonic acid, insulin, phenol red, sodium pyruvate and transferrin. The invention discloses another serum-free immune cell culture medium which further comprises cofactors including salidroside, luteolin and resveratrol. The invention discloses another serum-free immune cell culture medium which further comprises activators including an acetyl coenzyme A, a coenzyme Q10, glucose 6-phosphate and cyclic adenosine monophosphate. A part of embodiments of the invention can increase the positive ratio of cell phenotypes.

The invention discloses a tagatose-producing Bacillus subtilis genetically engineered bacterium and a method for preparing tagatose. The genetically engineered bacterium comprises constructing a single-expression or co-expression thermostable α-glucan phosphorylase, thermostable glucose phosphate Mutase, thermostable glucose phosphate isomerase, thermostable 6‑phosphate tagatose epimerase, and thermostable 6‑phosphate tagatose phosphatase. Starch can be effectively converted into tagatose by using the genetically engineered bacteria. Compared with the existing methods for producing tagatose, the method of the present invention has the advantages of being suitable for whole cell recycling, high safety performance, high yield, simple production process, low cost and easy large-scale preparation.

The invention discloses a coenzyme II NADP+ or NADPH content testing kit and its method. The kit comprises acid extractive fluid, alkali extractive fluid, mixed solution of HEPES and EDTA, glucose 6-phosphate, PMS, MTT, glucose 6-phosphate dehydrogenase solution, NaCl solution and mixed solution of ethanol and double distilled water. The method includes steps of extracting NADP+ and NADPH in a sample by acid and alkali extracting fluids on the basis of a spectrophotometric method; performing PMS hydrogen delivering action on NADPH, and reducing MTT to be formazan; detecting light absorption value under 570 nm, and testing the NADPH content; reducing the NADP+ to be NADPH by glucose 6-phosphate dehydrogenase, thereby detecting the NADP+ content. The kit and its testing method can effectively test the content of the coenzyme II NADP+ or NADPH, and are featured by high specifity, wide application range, high stability, and others.