响应面法优化大肠杆菌表达Hyperthermus butylicus耐热醛缩酶诱导条件的研究

Response Surface Optimization for Induction Conditions on Expression of the Thermophilic Aldolase from Hyperthermus butylicus in Escherichia coli

为了提高重组耐热2-脱氧核糖-5-磷酸醛缩酶的表达水平,该文基于中心组合实验的响应面分析方法,对基因工程E.coli的诱导条件进行了优化,具体考察因素包括诱导剂(异丙基-β-D-硫代半乳糖,IPTG)浓度和诱导时机.实验结果表明该方法可以提高目标蛋白的表达量,依据回归分析确定了最佳诱导条件为:IPTG浓度0.57mM,诱导时机OD600为0.76,理论最佳DERA的比活力为1.124u/mg,而实际平均比活力达到1.178u/mg,两者几乎相等,说明该模型与实际情况基本相符.

In order to improve the expression level of a recombinant thermophilie 2-Deoxyribose 5 phophate aldolase, the paper optimized the induction conditions in E. coil by the response surface methodology based on the central composite design. The factors included inducer concentration （raM. isopropyl β-D-thiogalactopyranoside, IPTG） and cell density （g L.） at the time of induction, The results show that this method can increase the expression quantity of protein, and determine the best induction condition, that is when IPIG density is 0.57 mG, and OD600 is 0.76. The maximal DERA specific activity was 1. 121 μ/mg which is nearly equal to the actual specific activity that is 1. 178 u/mg, this result indicates that the model corresponds with the actual sistuation.