工程蛋白转位和展开的蛋白质反应增强了人体PH-20分泌物在Pichia pastoris
Yue-Sheng Zhang1,2, Jin-Song Gong3, Jia-Yu Jiang1
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, People's Republic of China.
Applied microbiology and biotechnology
|January 4, 2024
概括
这项研究设计了Pichia pastoris以分泌人体氨酸酶,实现了4.9倍的产量增加. 这些方法通过协同翻译转位和展开的蛋白质反应激活来增强蛋白质分泌,用于工业应用.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 氨酸酶是降解氨酸 (HA) 的关键酶,在医学,化品和食品中越来越多的应用.
- 再组合氨酸酶为动物来源提供了可持续的替代品,需要高效的微生物生产系统.
- 皮奇亚牧师是一种广泛用于重组蛋白质生产的酵母宿主.
研究的目的:
- 在Pichia pastoris.中优化复合人体氨酶 (rhPH-20) 的分泌.
- 为增强蛋白质分泌而设计与内质网相关的分泌通路.
- 为了实现工业应用的 rhPH-20 的高水平生产.
主要方法:
- 在P. pastoris.中的一个构成性促进体下,人类氨酶的重组表达.
- 分泌途径的工程,包括信号优化,以实现协同翻译转位.
- 信号识别粒子组件的过度表达和通过ScHac1p.p激活未折叠蛋白反应 (UPR).
- 用于大规模生产的料批发发酵.
主要成果:
- 信号的优化提高了20%的分泌.
- 增强的同翻译转位和UPR激活导致分泌的氨酸酶活性增加2.1倍 (4.06 U/mL).
- 料批发发酵进一步提高了产量,达到19.82U/mL,总体增加了4.9倍.
结论:
- 结合的工程策略显著增强了P. pastoris.中的rhPH-20分泌.
- 这种方法,特别是增强同翻译转位,是改善酵母蛋白质分泌的新方法.
- 开发的策略适用于促进微生物宿主中其他有价值蛋白质的分泌.
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