多胺对毕赤酵母合成S腺苷甲硫氨酸的影响.pdf

现代食品科技 Modern Food Science and Technology 2012, Vol.28, No.12 多胺对毕赤酵母合成S-腺苷甲硫氨酸的影响 1,2 2 2 2 2 胡晓清 ，陈华新 ，储炬 ，张嗣良 ，庄英萍 （1.江南大学食品科学与技术国家重点实验室，江苏无锡 214122） （2.华东理工大学生物反应器工程国家重点实验室，上海 200237） 摘要：S-腺苷甲硫氨酸（S-adenosyl-L-methionine，SAM）是一种需求量巨大的活性氨基酸。SAM 生产主要是利用过表达甲硫 氨酸腺苷转移酶（methionine adenosyltransferase，MAT）的毕赤酵母（Pichia pastoris ）进行胞内合成，但较高密度培养条件下甲醇诱 导会造成一定程度的细胞死亡。本研究首先揭示甲醇对P. pastoris 造成过氧化损伤，然后分析了添加多胺（腐胺和亚精胺）对 SAM 合成影响及机制。结果揭示多胺可提高H O 酶活力，从而缓解细胞氧化损伤，此外还改善了MAT 酶活力和细胞生长，最终SAM 的 2 2 积累水平提高 19.2%。 关键词：毕赤酵母；S-腺苷甲硫氨酸；多胺；氧化损伤 文章篇号：1673-9078(2012)12-1617-1620 Influence and Mechanism of Polyamines Addition on S-adenosyl-L-methionine Synthesis by Pichia pastoris 1 2 2 2 2 HU Xiao-qing , CHEN Hua-xin , CHU Ju , ZHANG Si-liang , ZHUANG Ying-ping (1.State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122) (2.State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237) Abstract : S-adenosyl-L-methionine (SAM) was a type of active amino acid required largely. SAM was mainly produced through the biosynthesis reaction within Pichia pastoris cell that overexpressed methionine adenosyltransferase (MAT), while the inducer methanol led to cell death at some extent during cultivation process with high cell density. Thus this study firstly revealed that methanol exerted oxidative stress on P. pastoris cell, and then investigated the influences of adding polyamines (putrescine and spermidine) on SAM biosynthesis and reaction mechanism. The results revealed that p