通过表达调节元件修改和发酵优化,增强Bacillus subtilis中原粉降解α-amylase的细胞外生产
Dongbang Yao1,2,3,4, Xudong Han1,2,3,4, Huanhuan Gao1,2,3,4
1School of Life Sciences, Anhui University, Hefei, 230601, China.
Microbial cell factories
|June 28, 2023
概括
这项研究通过优化调节元素和发酵来增强Bacillus subtilis中的原粉降解α-氨酶 (RSDA) 生产. 该工程菌株实现了创纪录的高细胞外RSDA活性,为工业应用铺平了道路.
科学领域:
- 生物技术是生物技术.
- 酵素工程是什么意思 酵素工程
- 微生物发酵 微生物发酵
背景情况:
- 原粉降解α-氨酶 (RSDA) 在粉加工中节省成本,但由于生产水平低而受到影响.
- 在 Bacillus subtilis 中改善细胞外RSDA表达对于工业可行性至关重要.
研究的目的:
- 增强来自Pontibacillus sp.的原粉降解α-氨酶 (AmyZ1) 的细胞外生产. 在 Bacillus subtilis 中的 ZY.
- 优化表达调节元素和发酵条件,以提高RSDA产量.
主要方法:
- 促进体,信号和核糖体结合部位 (RBS) 序列的顺序优化.
- 构建一个双促进体系统 (Pveg-PylB) 和选信号 (SPNucB).
- 包括碳/来源和金属离子度在内的发酵优化.
主要成果:
- 与原始菌株相比,工程菌株WBZ-VY-B-R1实现了显著更高的细胞外AmyZ1活性.
- 摇瓶活动达到5733.5U/mL,3L发酵器活动达到49082.1U/mL.
- 这代表了再组合RSDA的最高报告的生产水平.
结论:
- 在Bacillus subtilis中实现了AmyZ1的创纪录的细胞外生产水平.
- 优化策略为工业RSDA应用提供了基础.
- 使用的方法为增强Bacillus subtilis中的其他蛋白质生产提供了有希望的方法.
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