重组毕赤酵母(Pichia pastoris)高产Lunasin的发酵工艺优化

Optimization for High Production Fermentation of Lunasin from Recombinant Pichia pastoris

Lunasin是一个最初从大豆中分离,具有43个氨基酸的多肽,具有抗高血压,抗氧化活性,预防癌症,抗炎和降胆固醇的活性。为了实现Lunasin的高效生产,对重组毕赤酵母GS115 LN诱导阶段甲醇补料策略(溶氧反馈补料,指数-恒速补料)以及诱导温度进行了优化,并且在此基础上对双碳源(山梨醇、甘露醇)和甲醇混合补料策略以及复合氮源与甲醇混合补料策略进行了优化。研究表明,最优补料策略以及诱导温度分别为指数-恒速补料、24℃,最终在流加1%大豆蛋白胨以及0.02%天冬氨酸条件下,Lunasin表达量最高,达到3.27 g/L,是单一甲醇诱导的1.27倍。

Lunasin is a 43 amino acid polypeptide originally isolated from soybean with bioactive properties such as antihypertensive, antioxidant activity, cancer prevention and therapy, anti-inflammation, hypocholesterolemic activity, anti obesity and immunomodulation. In order to achieve efficient production of Lunasin, the methanol fed methods （DO feedback fedding and index - constant speed feeding） and induction temperature of recombinant Pichia pastoris GSl15 LN were investigated. Furthermore, the feeding strategies of mixed carbon sources during induction phase were investigated. The results show that the best feeding strategies and induced temperature were index-constant speed feeding, 24 ℃. Consequently, the highest expression of Lunasin was 3.27g / L by feeding 1% soy peptone and 0. 02% aspartic acid,which was 1.27 times higher than the single methanol induction process.