plackettburman實验设计及响应值.doc

谷胱甘肽高产菌株的选育及其培养条件的研究 贾 建 萍 （浙江工业大学生物与环境工程学院 杭州 310014） 摘要：以编号为346的酿酒酵母为出发菌株，通过紫外线和60Coγ射线诱变处理，运用推理育种技术，选育到一株抗氯化锌和乙硫氨酸的突变株0.5Eth400-5。该菌株经摇瓶发酵谷胱甘肽产量为16.0mg/l，较出发株提高350%，每克干细胞含谷胱甘肽19.mg，较出发株提高318.6%。菌株经10次传代培养，谷胱甘肽产量下降10.7%，是一株性状较稳定可深入开发研究的优良菌株。在此基础上应用Plackett-Burman实验设计和响应面分析方法对菌体积累谷胱甘肽的培养条件进行了系统研究和优化，得到了1组优化的培养基。采用此优化培养基，谷胱甘肽产量达235.7mg/l，比优化前提高45.4%。在优化的培养条件下进行5L发酵罐试验，谷胱甘肽产量达638.9mg/l。BREEDING OF A GLUTATHIONE OVERPRODUCING STRAIN AND ITS CULTURE CONDITIONS Jia Jian-Ping (College of Biology and Environment Engineering, Zhejiang University of Technology, Hangzhou 310014) Abstract: A zinc chloride- and ethionine- resistant mutant 0.5Eth400-5 was obtained from its parent strain Saccharomyces cerevisiae ZG346 by UV and 60Coγ-ray treatment and rational screening. The glutathione productivity of the mutant reached 166.0mg/l by flask culture, which was 350.0% higher than that of the parent strain, and the glutathione content in the dried cells reached 19.8mg/g, which was 318.6% higher than that of the parent strain. A descend of only 10.7% in the glutathione yield of the mutant was observed after ten times of subculture. Therefore, the obtained mutant is a relatively stable strain that is worthy to be studied further. Systematic studies were carried out on the optimization of compositions of medium for accumulating the largest amount of glutathione in the strain. This optimizing medium was formulated vas Plackett-Burman design and response surface analysis (RSA). The glutathione yield was 235.7mg/l under the optimum condition,which was 45.4% higher than that of non-optimized medium. The glutathione yield was 638.9mg/l under the optimum condition by 5L fermenter culture. Key words: Glutathione, Saccharomyces cerevisiae , Strain breeding, Optimization 谷胱甘肽（Glutathione,简称GSH）是一种由谷氨酸、半胱氨酸和甘氨酸缩合而成的活性三肽。它广泛存在于动植物和微生物细胞中，是最主要的非蛋白巯基化合物，具有重要的生理功能。它可作为某些酶的辅酶，并对一些巯基酶有激活作用，是保护酶和其它蛋白质巯基的一种抗氧化剂，并且在氨基酸转运、维持红细胞膜的完整性、保护血红蛋白及解除毒物等方面具有重要的作用[1]。在临床上，谷胱甘肽是一种治疗肝脏疾病、肿瘤、中毒、白内障、衰老及变态反