一个紧的,通用的融合标签使粉生物转化和优化Bacillus subtilis中代谢的挑战性酶的强有力的表达成为可能
Yuqiao Wang1, Jinlong Li2, Wenhui You1
1School of Biological Engineering, Dalian Polytechnic University, Dalian, China; Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China; National Technology Innovation Center of Synthetic Biology, Tianjin, China.
研究人员开发了一种新的N30微标签,用于生物技术中高效的酶表达. 这种标签显著增强了蛋白质分泌,并使Bacillus subtilis能够达到以前无法实现的表达水平,从而推进可持续的生物制造.
科学领域:
- 生物技术是生物技术.
- 蛋白质工程是指蛋白质工程.
- 代谢工程是代谢工程.
背景情况:
- 在多酶系统中酶蛋白的有效表达对于可持续生物技术至关重要,但仍然具有挑战性.
- 在Bacillus subtilis中高水平的葡萄糖-6-酸盐异构酶表达激发了新的蛋白质表达策略.
研究的目的:
- 开发一种新的,高效的,可扩展的方法来表达Bacillus subtilis中的酶蛋白.
- 识别和表征一种新的N端微标签,用于增强蛋白质分泌和表达.
主要方法:
- 基于三维蛋白质结构的区域截断分析被用于识别30氨基酸N终端微标记 (N30).
- 应用N30标签用于人工粉合成途径 (ASAP) 中的酶表达.
- 为了评估N30的效率,采用了料批发发酵和融合表达策略.
主要成果:
- 在ASAP中,N30标签使八种关键酶的分泌水平达到大约10μg/L.
- 一个修改的N30标签 (N30(M22D) -TpiA) 在料批发酵中达到15μg/L的分泌水平.
- 在Bacillus subtilis中,N30促进了以前无法表达的真核蛋白的可检测表达.
- 限制速率酶的融合表达增加了利博弗拉和menaquinone-7标位,分别是2.4倍和2.3倍.
结论:
- 标签N30是一个紧的,活动的保存,和可扩展的解决方案,用于Bacillus subtilis的酶合成.
- N30显著增强了用于可持续二氧化碳转化和粉生物转化应用的分泌表达系统.
- 这项技术在蛋白质合成和代谢工程方面展示了广泛的多功能性和效率.
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