优化早期分泌途径的多种组合,以增强Komagataella phaffii中的细胞外蛋白分泌
Hai Xu1,2,3, Shijie Wang1,2,3, Kang Li1,2,3
1School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
Journal of agricultural and food chemistry
|July 5, 2025
概括
通过优化途径来改进甲基营养酵母 Komagataella phaffii (K. phaffii) 的工程改进了蛋白质分泌. 这提高了工业酶和治疗性蛋白质的产量.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 甲基营养酵母 Komagataella phaffii (K. phaffii) 是异质蛋白质生产的关键平台.
- 在K. phaffii中低效的分泌贩运限制了其用于生产酶和生物制剂等蛋白质的工业可扩展性.
研究的目的:
- 系统地设计K. phaffii的早期分泌途径,以提高细胞外产量.
- 改善真菌α-氨酶 (AmyA),细菌β-葡萄糖酶 (BgA) 和人血清白蛋白 (HSA) 的分泌.
主要方法:
- 综合分泌工程与3'未翻译区域 (3'UTR) 优化,以改善共翻译ER转位.
- 实施多层干预措施,包括ER膜扩张,ER相关降解 (ERAD) 调制和囊泡运输增强.
- 应用了蛋白质特定的优化策略,以最大限度地提高细胞外蛋白质产量.
主要成果:
- 分泌通路工程使细胞外活动增加了1.3-3.4倍.
- 多层干预可以将细胞外蛋白水平提高1.8-4.0倍.
- 在蛋白质分泌方面取得了5.2-6.4倍的累积改善,最佳策略因蛋白质而异.
结论:
- 建立了一个模块化和定制的工程框架,以提高K. phaffii. 的分泌效率.
- 这种方法广泛适用于改善工业酶和治疗生物制剂的生产.
- 优化的K. phaffii菌株为可扩展的生物制造提供了显著的潜力.
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