36 results about "Pichia garciniae" patented technology

The invention provides a method for producing, efficiently secreting and expressing core fucose-base transferases FUT8 in pichia pastoris expression systems, and belongs to the field of genetic engineering and the field of protein engineering. The method mainly comprises constructing the pichia pastoris secretion and expression systems for the recombinant human-derived core fucose-base transferases FUT8; separating and purifying the recombinant human-derived core fucose-base transferases FUT8; detecting the enzymatic activity of the recombinant human-derived core fucose-base transferases FUT8. The method has the advantages that the core fucose-base transferases FUT8 synthesized, secreted and expressed by the aid of the method can be widely applied to protein inhibitor, biological function and in-vitro oligosaccharide research; the method is low in synthesis cost, short in synthesis period and easy to technically implement, and the core fucose-base transferases FUT8 obtained by the aid of the method are high in purity.

The invention discloses a synthesized duck beta-defensins-2 gene, a recombinant plasmid containing the gene as well as production methods for a recombinant yeast transformant and a recombinant duck beta-defensins-2 protein. The recombinant duck beta-defensins-2 gene refers to the cDNA (complementary Deoxyribonucleic Acid) gene sequence of a duck AvBD2 gene registered in GenBank; meanwhile, the nucleotide sequence in a mature peptide segment of the duck AvBD2 gene is transformed according to the preference of a yeast codon; the duck beta-defensins-2 gene is synthesized by using a gene synthetic method; the construction process of the recombinant plasmid containing the gene sequentially comprises the following steps of: firstly, setting an upstream primer and a downstream primer of the duckbeta-defensins-2 gene; secondly, carrying out PCR (Polymerase Chain Reaction) amplification on a duck beta-defensins-2 gene segment; and thirdly, constructing a recombinant plasmid pPICZalpha-A-AvBD2. The production method for the recombinant duck beta-defensins-2 protein comprises the following steps of: firstly, preparing and linearizing the recombinant plasmid; secondly, preparing a competent cell obtained by electrotransformation of pichia pastoris; and thirdly, electroporating the pichia pastoris. The production method for the recombinant duck beta-defensins-2 protein comprises the step of induced expression of the positive yeast transformant. Compared with the prior art, synthesized duck beta-defensins-2 gene, the recombinant plasmid and the production methods disclosed by the invention have the advantages of activities of resisting bacteria and promoting the growth, suitability for application to the livestock production, low production cost and high production efficiency.

The invention discloses a Pichia pastoris CH01, a fermenting cultivating method thereof and an application. The preservation number of the Pichia pastoris CH01 is CGMCC No.4273. The fermenting cultivating method of the Pichia pastoris CH01 comprises the steps of: 1) inoculating activated Pichia pastoris CH01 into the frequently used fermenting culture solution of the Pichia pastoris CH01 so as tocultivate seed solution and 2) inoculating the seed solution obtained in the step 1) to the frequently used fermentation medium of the Pichia pastoris CH01 so as to aerobically culture and obtain thePichia pastoris CH01 solution. The Pichia pastoris CH01 has short fermentation period, is simple in preparation process, and has reliable industrial production. The invention has wide application prospect in preventing and treating potato late blight and controlling the postharvest diseases of fresh fruit and vegetables.

The invention discloses an alkaline pectinase secretion-enhanced strain and an application thereof, and belongs to the technical field of genetic engineering. Through a gene recombination technology, UPR related genes of pichia pastoris are cloned and linked to a pichia pastoris expression vector pPGAZA and are converted to a Pichia pastoris GS115-pPIC9K-PGL strain, so that a plurality of strains, namely GS115-PGL / pGAPZA-HAC1, GS115-PGL / pGAPZA-ERO1, GS115-PGL / pGAPZA-UBC1 and the like, which are stronger in secretion and expression of alkaline pectinase than an original strain, are obtained, the enzyme activities of the strains, before the application of the method, are improved by 24.1%, 35.5% and 22.3%, and with the significant improvement, a good foundation is laid for the large-scale production of the alkaline pectinase. The alkaline pectinase disclosed by the invention can catalyze the pyrolysis of an alpha-1,4 glucosidic bond of polygalacturonic acid by virtue of a trans-elimination effect under an alkaline condition;and the alkaline pectinase can be widely applied to such industries as food, textile, paper-making and the like.

The invention provides a construction method for producing fumaric acid based on pichia stipitis synthetic strain fermented xylose. The method comprises the following steps: using pichia stipitis having a natural xylose metabolic capability as a host; deleting genes PSfum1 and PSfum2 that encode fumarase in tricarboxylic acid cycle of pichia stipitis, to acquire fumarase deficient mutants; heterologously expressing a modified fumaric reduction synthesis module on the basis of the acquired mutants, using xylose to ferment yeast synthesis strains of fumaric acid; on the basis of the yeast synthesis strains, expressing translocator YMAE that is modified with codons and originated from Chestnut wine fission yeasts, to acquire pichia stipitis synthesis strains for producing fumaric acid by fermented xylose; efficiently fermenting the acquired strains in a fully synthesized fermentation culture medium to produce fumaric acid by fermentation of xylose; cultivating by fermentation for 3-4 days at a temperature of 30 degrees centigrade with a stirring speed of 150-200rpm. The content of fumaric acid is greater than 4g / L.

The invention discloses a laccase LbLCC9I sequence capable of being expressed in Pichia pastoris and Arabidopsis thaliana after being optimized and application thereof. The nucleotide sequence of the laccase LbLCC9I is disclosed as SEQ ID NO 1, the full-length is 1518bp, and the sequence of the coded protein is disclosed as SEQ ID NO 2 and is composed of 501 amino acids. After being transformed into Pichia pastoris and Arabidopsis thaliana, the sequence can express the active laccase. The laccase can be used for dye decolorization.