优化培养条件和裂解方法提高hRANKL在大肠杆菌内的表达

Enhancing hRANKL production in Escherichia coli by optimizing culture conditions and lysing program

摘要:

RANKL/RANK/OPG轴在骨代谢过程中起到中心调节作用，也是近年来骨相关疾病治疗研究的热点之一。RANKL蛋白在RANKL/RANK/OPG轴信号传递过程中起到关键作用，在骨代谢相关实验研究中用途广泛。但是，使用大肠杆菌Escherichia coli可溶表达重组人源RANKL蛋白 (hRANKL) 时产量远低于鼠源RANKL (mRANKL)。本研究通过将LB培养基pH值调整并稳定在7.5、降低诱导表达温度至16 ℃并优化细菌裂解条件，成功地将可溶hRANKL产量增加到了对照组的5?12倍。该方法有效提高了hRANKL在大肠杆菌中可溶表达的产量，同时也是研究重组蛋白在大肠杆菌内的可溶表达策略的有益尝试。

Abstract:

RANKL/RANK/OPG axis is important in bone metabolism regulation, and becomes a popular research area in bone diseases. RANKL is a critical part of RANKL/RANK/OPG axis, and widely required in bone metabolism research. However, the yield of recombinant soluble human RANKL (hRANKL) in Escherichia coli is much lower than mouse RANKL (mRANKL). In this study, by adjusting and stabilizing the pH value of LB medium at 7.5, lowering the inducing temperature to 16 ℃ and optimizing the lysis program, the yield of soluble hRANKL increased by approximately 5 to 12-fold over the non-adjusted group. Our experiment effectively enhanced soluble hRANKL expression in E. coli and might constitute a meaningful attempt to obtain soluble expression of recombinant protein in E. coli.