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

Author: Luyang CHE ¹ ; Zhilan GUO ² ; Qinghua GUO ¹ ; Bing PAN ³ ; Jingzhe LI ² ; Changzhen LIU ² ; Peng HUANG ¹
Author Information

1. Department of Orthopedics, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
2. Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China.
3. Beijing Jiquan Biology Technology Co., LTD., Beijing 100070, China.
Publication Type:Journal Article
Keywords: Escherichia coli; RANKL; osteoporosis; protein expression
From: Chinese Journal of Biotechnology 2017;33(5):849-862
CountryChina
Language:Chinese
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.