在Saccharomyces cerevisiae中的重组猪表皮生长因子的分泌表达和发酵优化
Jie Liu1, Wenjun Xu2, Bingbing Xia2
1College of Biotechnology and Pharmaceutical Engineering, West Anhui University, Lu'an, Anhui, 237012, China.
Protein expression and purification
|October 9, 2025
概括
我们开发了一种酵母菌株,用于生产用于猪料的重组猪皮表皮生长因子 (PoEGF). 这种具有成本效益的方法可以在不经过净化的情况下产生生物活性PoEGF,从而提供了抗生素替代品.
科学领域:
- 生物技术是生物技术.
- 动物科学动物科学
- 分子生物学分子生物学
背景情况:
- 猪皮表皮生长因子 (PoEGF) 支持小猪肠道健康和免疫力.
- 目前的重组PoEGF (rPoEGF) 生产是昂贵的,需要大量的净化.
- 在生猪生产中,rPoEGF为料抗生素提供了潜在的替代品.
研究的目的:
- 开发一种具有成本效益和可扩展的方法来生产生物活性rPoEGF.
- 为了设计Saccharomyces cerevisiae以有效分泌rPoEGF.
- 为了优化发酵条件的高产 rPoEGF生产.
主要方法:
- 构建了一个复合酵母菌株 (Saccharomyces cerevisiae INVSc1) 与一个α-因子信号-PoEGF-6×他的表达盒.
- 通过SDS-PAGE和西方 blot.确认蛋白质表达.
- 在4L生物反应器中优化了两阶段发酵策略 (食批量生长,在25°C,pH为5.0,DO为30%的温度下引起银河糖的表达).
- 使用BALB/c 3T3纤维细胞增殖试验验验证的生物活性.
主要成果:
- 达到了最大的10963个单位/毫升的线粒生成活性.
- 达到高细胞密度培养 (OD600 ≈ 71.5).
- 获得了30.2 mg/L的最终分泌蛋白产量.
- 证明了培养上游物作为料添加剂的直接应用,无需净化.
结论:
- 开发了一种实用,可扩展和具有成本效益的方法,用于使用工程酵母生产生物活性rPoEGF.
- 含有rPoEGF的酵母培养物可以直接用作功能性料添加剂.
- 这种方法支持rPoEGF作为安全的,无抗生素的生长促进剂在猪生产中的应用.
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