Recombinant pichia pastoris for producing human casein macropeptide and application of recombinant pichia pastoris

• Main Authors: CHEN JIAN, LIU YANFENG, PLUGOC CHENG, LI JIANGHUA, LIU LONG, LYU XUEQIN, DENG MENGTING
• Format: Patent
• Language: Chinese; English
• Published: 28.03.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention relates to recombinant pichia pastoris for producing human casein macropeptide and application of the recombinant pichia pastoris, high-copy expression of human casein macropeptide genes is realized in a pichia pastoris strain, and secretory expression of the human casein macropeptide is realized. In a product produced by the constructed recombinant pichia pastoris, the content of impure protein is low, subsequent purification is facilitated, the shake flask yield is up to 234 mg/L, fusion expression is not needed in the process, the impure protein in fermentation supernate is few, the secretory expression quantity of target protein accounts for 93% of the total secretory protein of the pichia pastoris, and the yield is high. The method lays a foundation for efficiently producing breast milk protein in pichia pastoris metabolic engineering. 本发明涉及一种生产人酪蛋白巨肽的重组毕赤酵母及其应用，在毕赤酵母菌株中高拷贝表达人酪蛋白巨肽基因，实现人酪蛋白巨肽的分泌表达。本发明通过构建的重组毕赤酵母生产的产物中，杂蛋白含量低，便于后续纯化，摇瓶产量即高达234mg/L，该过程无需融合表达，且发酵上清液杂蛋白少，目的蛋白的分泌表达量占毕赤酵母总分泌蛋白的93