重组毕赤酵母表达蛋清溶菌酶发酵条件的优化  

Optimization of fermentation conditions for expression of egg white lysozyme by recombination Pichia pastoris

作　　者：孙玮遥 宋增健 王向东[2] 林剑[1] 

机构地区：[1]烟台大学生命科学学院,山东烟台264005 [2]山东大学医学院,山东济南250100

出　　处：《中国酿造》2017年第2期54-57,共4页China Brewing

基　　金：国家自然科学基金(81370881);烟台大学研究生科技创新基金重点项目(YDZD1610)

摘　　要：该研究以甲醇诱导型重组毕赤酵母(Pichia pastoris)NCY-2为研究菌株,在摇瓶水平上首先考察了诱导时间、甲醇含量、诱导p H及诱导温度对蛋清溶菌酶表达的影响,然后通过正交试验设计优化出了该菌株的最佳发酵条件,并进一步研究了摇瓶发酵过程中的菌体生长和酶活力随时间的变化规律。研究结果表明,蛋清溶菌酶的最适诱导时间为96 h,甲醇含量为2%,诱导p H值为3.5,诱导温度为20℃;在此条件下发酵液的蛋清溶菌酶酶活力达到775 U/m L,是优化前的2.2倍。Using methanol inducible recombinant Pichia pastoris NCY-2 which produces egg-white lysozyme （EWL） as experimental strain, the effect of induction time, methanol concentration, induction pH and temperature on the expression of EWL were investigated by single factor experiments in shake flasks and the optimum fermentation conditions were obtained through orthogonal experiment design. The variation of cell growth and EWL activity with time in shake flask fermentation was further studied. The results showed that the optimal conditions were induction time 96 h, methanol concentration 2%, induction pH 3.5 and temperature 20 ℃. The EWL activity reached 775 U/ml under the optimized conditions, which was 2.2 times compared with that before optimization.

关 键 词：蛋清溶菌酶 毕赤酵母 发酵条件 优化 

分 类 号：Q815[生物学—生物工程]