摘要
以重组毕赤酵母GS115/pPICZaA-Prochy为凝乳酶生产菌种,研究其在5L发酵罐水平的最佳产酶条件,为中试生产提供依据。采用批量发酵,甲醇浓度0.1%,诱导阶段每12h添加10g/L蛋白胨,发酵液pH值为3.0,油酸浓度0.2%,诱导初期批量加入10g/L山梨醇共基培养,诱导84h可获得152SU/mL的酶活力。采用十二烷基磺酸钠-苯琼脂糖凝胶电泳分析,在发酵液(去菌体后)样品盐离子浓度2mol/L,流速2.5mL/min,酶回收率达到82%,浓缩倍数20倍。
The optimal conditions for producing chymosin in 5L fermenter by recombinant Pichia pastor/s Gsl 15/pPICZaA-Proehy were determined in order to offer technical bases for pilot scale production. After induction for 84h, the highest enzyme activity （152SU/ml） was achieved with batch culture, in which, 10g/L sorbitol was added at the beginning of induction phase, methanol concentration was kept at 0.1%, 10g/1 peptone was added every 12h, pH value was kept at 3.0 and the concentration of oleic acid was 0.2%. The purification of chymosin was analyzed by sodium dodecyl sul- fate benzene agarose gel electrophoresis （SDS-PAGE）. Under the concentration of salt ions at 2mol/L and flow rate 2.5mL/min, the recovery rate of chymosin was 82.33% and the enzyme sample was concentrated by 20 folds.
出处
《中国酿造》
CAS
北大核心
2011年第10期33-37,共5页
China Brewing
基金
国家高技术研究发展计划项目863计划(2006AA10Z306)
吉林省科技发展计划项目(20060219,20080228)
现代农业产业技术体系建设专项资金资助(农科教发[2007]14号)资助
关键词
凝乳酶
毕赤酵母
发酵条件
纯化
chymosin
Pichia pastoris
fermentation conditions
purification
