在Bacillus subtilis中,一种基于T7促进剂的高表达性异质蛋白质的简单而强壮的T7促进剂
Jing Ye1, Yunjie Li2, Yuqing Bai2
1Tianjin Key Laboratory of Food Science and Biotechnology, College of Biotechnology and Food Science, Tianjin University of Commerce, Tianjin, China.
Bioresources and bioprocessing
|April 22, 2024
概括
我们使用T7促进器技术开发了一种新的Bacillus subtilis表达系统. 该系统实现了高重组蛋白产量,促进了工业应用.
科学领域:
- 生物技术是生物技术.
- 微生物工程 微生物工程
背景情况:
- 在大肠杆菌中,T7促进体系统被广泛用于蛋白质表达.
- 细菌细菌是一种具有工业相关性的微生物,具有重组蛋白质生产的潜力.
研究的目的:
- 在Bacillus subtilis中开发一种基于易于T7促进体的蛋白质表达系统.
- 为增强蛋白质表达而设计B. subtilis菌株和兼容的质粒.
主要方法:
- 通过整合T7RNA聚合酶基因并淘汰蛋白酶和其他非必需基因,构建了一个新的B. subtilis菌株 (SCK22).
- 开发了一种具有T7促进体和B. subtilis核糖体结合部位 (RBS) 的等离子体 (pHT7),用于基因表达.
- 在工程B. subtilis菌株中表达了多种重组蛋白质,包括绿色光蛋白和酶.
- 进行高细胞密度食批发发酵,以评估生产能力.
主要成果:
- 使用SCK22/pHT7系统实现了重组蛋白的高表达水平 (25-40%的总细胞蛋白).
- 通过料批发发酵成功生产高达4.78g/L的内醇单酸酶.
- 对各种重组蛋白具有广泛的应用性.
结论:
- 开发的B. subtilis T7表达系统提供了高蛋白产量和遗传稳定性.
- 该系统适用于高细胞密度发酵和工业规模的重组蛋白质生产.
- 该系统提供了诸如简单的基因操作和高转化效率等优势.
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