在Komagataella phaffii中具有三环结构的III型重组人体原的有效分泌表达
Yaqian Ma1,2, Yang Li3, Nan Wang1,2
1Department of Biological Engineering, College of Life Sciences, Yantai University, Yantai, 264005, China.
Applied microbiology and biotechnology
|September 3, 2025
概括
我们开发了一种新的方法, 这种可扩展的平台实现了高产量,为功能性原蛋白的工业和临床应用铺平了道路.
科学领域:
- 生物技术
- 生物化学
- 材料科学
背景情况:
- 再组合人体原 (rhCol) 具有高纯度和低免疫性,可用于生物医学.
- 在结构稳定和功能活跃的 rhCol 大规模生产方面仍然存在挑战.
研究的目的:
- 在Komagataella phaffii中开发一个高效且可扩展的 rhCol生产平台.
- 优化原蛋白变体并确定有效的微生物prolyl-4-hydroxylases (P4H) 以提高产量.
主要方法:
- 使用链间盐桥工程进行五种III型原蛋白变体 (ColP1-ColP5) 的合理设计.
- 对微生物P4H进行查,确定Bacillus megaterium P4H (BmP4H) 是氧化的最佳基因.
- 优化酵母菌株,促进剂 (PAOX1),信号,并将发酵量扩大到5L.
主要成果:
- 科尔P2变体表现出优异的表达和功能.
- 在酵母中,BmP4H的同表达使得稳定的三环螺旋形成.
- 获得了2.54g/L的高rhCol产量,并确认了三螺旋结构.
结论:
- 建立了一个集成和可扩展的高水平功能rhCol生产平台.
- 这种方法为复合的工业和临床应用提供了基础.
- BmP4H的同表达是酵母中稳定的三环原蛋白的关键.
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