响应面法优化重组大肠杆菌PUCRF发酵培养基.pdfVIP

※生物工程 食品科学 2011, Vol. 32, No. 09 225 响应面法优化重组大肠杆菌PUCRF 发酵培养基 1,2 1 1 1 1, 王 华 ，于寒松 ，朴春红 ，刘俊梅 ，胡耀辉 * (1.吉林农业大学食品科学与工程学院，吉林长春 130 118；2 . 内蒙古民族大学生命科学学院，内蒙古通辽 028000) 摘 要：为进一步提高重组大肠杆菌( PUCRF)产β- 环糊精葡萄糖基转移酶(β-CGTase) 的酶活力，以构建的一株能 在胞外分泌表达β-CGTase 工程菌E.coli PUCRF 为出发菌株，利用响应面法对其发酵工艺条件进行优化。根据Box- Benhnken 中心组合设计原理，在单因素试验基础上采用四因素三水平的响应面分析法确定重组大肠杆菌PUCRF 最 佳培养基组成(g/ 100mL)为：玉米淀粉 0.962 、玉米浆0 .097 、Mg SO4 ·7H2 O 0 .153 、K2HPO4 ·3H2 O 1.2 35 。经 过优化，β-CGTase 产量可达36 10U/mL ，是初始条件下β-CGTase 酶活力(2690U/mL) 的1.34 倍。 关键词：响应面分析；β- 环糊精葡萄糖基转移酶；发酵；培养基 Optimization of Culture Medium for Recombinant Escherichia coli PUCRF Using Response Surface Analysis 1,2 1 1 1 1, WANG Hua ，YU Han-song ，PIAO Chun-h ong ，LIU Jun-m ei ，HU Yao-hui * (1. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130 118, China ； 2. College of Life Science, Inner Mongolia University for the Nationalities, Tongliao 028000, China) Abstract ：An engineered strain E. coli PUCRF producing β-cyclodextrin glycosyltransferase ( β-CGTase) has constructed in our lab . In this study, response surface analysis was used to optimize the culture medium composition for the production of β-CGTase by the strain. On the basis of one-factor-at-a-time experiments, a 4-factor, 3-level central composite design was performed. The optimum culture medium composition (g/ 100 mL) was determined as follows: corn starch 0.962, corn syrup 0.097, MgSO4 ·7H2 O 0.153 and K2HPO4 ·3H2O 1.235. The corresponding β-CGTase yield was 36 10 U/mL, 1.34 times higher than before the optimization (26