D-乳糖醇代替异丙基-β-D-巯基半乳糖苷对重组蛋白诱导表达的影响

目的:用D-乳糖醇代替异丙基-β-D-巯基半乳糖苷(isopropyl-β-D-thiogalactoside,IPTG)作为诱导剂,诱导重组葡萄糖苷酶的表达。方法:首先通过单因素试验探讨D-乳糖醇浓度、诱导时间及诱导温度对重组葡萄糖苷酶表达的影响,从而确定最佳影响因素的水平值,然后在单因素试验的基础上采用三因素三水平的响应面法对重组葡萄糖苷酶表达进行优化分析,依据回归分析方法确定最佳诱导工艺条件。结果:最佳诱导工艺条件为D-乳糖醇的诱导终浓度1.0 mmol/L、诱导时间7.0 h、诱导温度34℃。结论:D-乳糖醇可以代替IPTG作为一种高效的lac及其衍生物的启动子基因工程重组蛋白的理想诱导剂。

D-Lactitol as a Substitute for Isopropyl-β-D-Thiogalactoside to Induce Recombinant Protein Expression

Objective: To induce the expression of recombinant glycosidase using D-lactitol instead of isopropyl-β-Dthiogalactoside
(IPTG). Methods: Single-factor experiments were performed to discuss the effect of D-lactitol concentration,
induction time and temperature on the expression of recombinant glucosidase and consequently select three appropriate
levels for each of the three variables. Subsequently, the levels of these variables were optimized by regression analysis and
response surface methodology. Results: The optimal induction conditions were determined to be induction at 34 ℃ for
7.0 h with 1.0 mmol/L lactitol. Conclusion: D-Lactitol can replace IPTG as an ideal inducer for the production of
recombinant proteins in genetically engineered strains with the efficient promoter of lac and its derivatives.