在Saccharomyces cerevisiae中表达Enterocin A
Michelle Rossouw1, Gerhardt Coetzee2, Rosemary A Cripwell1
1Department of Microbiology, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa.
Probiotics and antimicrobial proteins
|August 30, 2025
概括
在Saccharomyces cerevisiae酵母中重新组合表达素A是一种有前途的大规模生产方法. 这项研究强调了酵母是细菌素的可行的宿主,通过辅助蛋白共同表达可以改善植物423.
科学领域:
- 生物技术
- 微生物学
- 分子生物学
背景情况:
- 二级细菌素是乳酸细菌产生的抗菌.
- 在Saccharomyces cerevisiae中,重组表达为这些细菌素提供了可扩展的生产系统.
- 对细菌素活性而言,二硫化键的形成至关重要,原生操作子通常包括辅助蛋白来促进这一过程.
研究的目的:
- 为了比较Saccharomyces cerevisiae中代码优化的素A,植物素423和世界素ST4SA的复合表达.
- 评估这些重组的细菌素活性和翻译后修饰.
- 评估S. cerevisiae作为大型细菌素生产的主体的潜力.
主要方法:
- 在S. cerevisiae中表达了对素A (EntA_Opt),植物素423 (PlaX_Opt) 和世界素ST4SA (MunX_Opt) 的代优化基因.
- 使用振动瓶和批次发酵量化的表达水平.
- 使用Nano-LC-MS/ MS评估了细菌素的活性,并分析了翻译后的修饰,包括二硫化键的形成.
主要成果:
- 与PlaX_Opt和MunX_Opt相比,震动瓶和批发发酵产生了更高的EntA_Opt水平.
- 与植物素423相比,内素A和世界素ST4SA的细菌素活性更高.
- 与PlaX_Opt相比,纳米LC-MS/MS分析显示了更多的EntA_Opt的正确二硫化键构造.
结论:
- 麦芽是一种有前途的宿主,可用于IIa类细菌素的重组生成,特别是素A.
- 复合植物素423的活性降低可能是由于它在S. cerevisiae中缺少原生辅助蛋白 (PlaC).
- 应研究辅助蛋白的同时表达,以增强复合植物423.
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