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在大肠杆菌Escherichia coli中,重组人类骨形态遗传蛋白-2的高水平可溶性表达

Youming Cui¹, Yanqiu Liang¹, Bin Zhang¹

¹The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, P. R. China.

ACS synthetic biology

|September 9, 2025

概括

研究人员开发了一种生产可溶性人类骨型遗传蛋白-2 (hBMP-2) 在大肠杆菌中的方法. 这一突破克服了纳入体的形成,为工业规模的hBMP-2生产铺平了道路.

科学领域:

生物技术是生物技术.
蛋白质工程是指蛋白质的工程.
分子生物学分子生物学

背景情况:

人类骨型基因蛋白-2 (hBMP-2) 对于骨和软骨的形成至关重要.
hBMP-2的工业应用受到其在 prokaryotic 系统中形成包容体的倾向的限制.

研究的目的:

在大肠杆菌中建立复合hBMP-2 (rhBMP-2) 表达系统.
为了提高rhBMP-2的可溶性,用于工业规模生产.

主要方法:

利用pCold II等离子体和SHuffle T7菌株进行rhBMP-2表达.
研究的策略包括与合成内在无序蛋白质 (SynIDPs) 和标签的分子伴侣共同表达,分泌表达和融合表达.
采用西方斑点分析和金属酸盐染色学来对蛋白质进行表征和净化.

主要成果:

与分子陪伴物pGro7的同时表达改善了rhBMP-2的溶解性.
与SynIDPs的融合导致了完全的单体溶解性.
GB1标签实现了完全溶解的rhBMP-2表达,产生单体和二元形式.
纯化的rhBMP-2达到了109.7±5.0毫克·L-1的表达水平.

结论:

在大肠杆菌中成功证明完全溶解的rhBMP-2表达.
开发了有效的策略,以克服在重组蛋白质生产中的纳入体形成.
为工业规模生产可溶性 rhBMP-2 提供了基础.

关键词:

埃舍里奇亚大肠杆菌 (Escherichia coli) 是一个大肠杆菌.人类骨形态遗传蛋白-2 小分子标签小分子标签可溶性表达的可溶性表达

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