69 results about "Yeast display" patented technology

The invention discloses a method for catalytic synthesis of vitamin E succinat by utilizing yeast display lipase, and the method comprises the following steps: dissolving vitamin E and succinic anhydride in an organic solvent, adding the yeast display lipase, reacting at 50-60 DEG C for 10-14 hours under an anaerobic condition, and separating and purifying, thus obtaining the vitamin E succinate.The preparation method of the yeast display lipase comprises the following steps: transforming the recombinant plasmid subjected to linear treatment into a Pichia Pastoris yeast GS115, inoculating the obtained transformants into a BMMY (buffered methanol-complex medium), inducing and culturing for 72-144 hours, carrying out centrifugal collection on thallus, washing, and carrying out organism printing and frozen drying on the washed thallus, thus obtaining the yeast display lipase. Through displaying lipase outside cells, and utilizing the lipase to conduct catalytic synthesis of the vitamin E vitamin E succinate, the conversion efficiency can be improved, the reaction time is shortened, and the production cost is lowered.

The invention relates to preparation of high-activity phospholipids enzyme D and cell surface display phospholipids enzyme D yeast whole cell catalysts, which belongs to a method for carrying out site directed mutagenesis wild phospholipids enzyme D by recombinant deoxyribonucleic acid (DNA) for improving the activity, connecting the mutated gene with yeast show carriers pPIC9K-Flo and efficiently displaying the mutated gene on the surface of pichia pastoris cells, and relates to a preparation method of the high-activity phospholipids enzyme D and cell surface display phospholipids enzyme D yeast whole cell catalysts. The method has the advantages that the activity of the wild phospholipids enzyme D is improved and is shown on the surfaces of the pastoris cells, the stability is improved,and the advantages of immobilized enzymes are realized. The method has the technical scheme that wild phospholipids enzyme D genes are separated from microbes, particularly streptomyces aureofuscus, the mutation is carried out on the amino acid residue of Glu69 and Ser285, through the efficient expression on the surfaces of the pichia pastoris cells, the enzyme activity of the high-activity phospholipids enzyme is improved by 11 percent than the wild type phospholipids enzyme D, and the enzyme activity of the recombination strains GS115 / pPIC9K-Flo-pldm prepared through high-density fermentation is 120U / (g. stem cells).

The invention discloses a method for catalytic synthesis of L-ascorbic acid linoleate by utilizing yeast display lipase, and the method comprises the following steps: dissolving L-ascorbic acid and linoleic acid in an organic solvent, adding the yeast display lipase, reacting at 50-60 DEG C for 4-8 hours under an anaerobic condition, and separating and purifying, thus obtaining the L-ascorbic acid linoleate. The preparation method of the yeast display lipase comprises the following steps: transforming the recombinant plasmid subjected to linear treatment in a Pichia Pastoris yeast GS115, inoculating the obtained transformants into a BMMY (buffered methanol-complex medium), inducing and culturing for 72-144 hours, carrying out centrifugal collection on thallus, washing, and carrying out organism printing and frozen drying on the thallus, thus obtaining the yeast display lipase. Through displaying lipase outside cells, and utilizing the lipase to conduct catalytic synthesis of the L-ascorbic acid L-ascorbic acid linoleate, the conversion efficiency can be improved, the reaction time is shortened, and the production cost is lowered.