在Pichia pastoris中研究大卜过氧化酶的高效表达
Yaping Wang1,2, Yidan Jing1,2, Weizhen Li1,2
1National Biopesticide Engineering Technology Research Center, Hubei Biopesticide Engineering Research Center, Hubei Academy of Agricultural Sciences, Biopesticide Branch of Hubei Innovation Centre of Agricultural Science and Technology, Wuhan 430064, China.
Molecules (Basel, Switzerland)
|November 27, 2025
概括
这项研究优化了在Pichia pastoris中使用分子陪伴剂优化的过氧化酶 (HRP) 生产. 与PDI1和HAC1的同时表达显著提高了HRP产量和活性,在大规模发酵中达到创纪录的水平.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 微生物发酵 微生物发酵
背景情况:
- 卜过氧化酶 (HRP) 是一种具有多样性应用的关键血氧化还原酶.
- 高效的HRP生产对于其在生物技术和诊断领域的广泛使用至关重要.
研究的目的:
- 为了增强Pichia pastoris.中的HRP生产.
- 研究分子伴侣对HRP表达和活性的影响.
主要方法:
- 构造的Pichia pastoris菌株具有不同的HRP-C基因拷贝数.
- 通过使用不同的诱导方法与分子伴侣 (PDI1,HAC1,BIP1) 共同表达HRP-C.
- 在50L发酵器中进行高密度发酵.
主要成果:
- 与PDI1和HAC1共同表达的3个副本的HRP-C Pichia pastoris菌株显示HRP产量 (1.4倍) 和活性 (1.2-1.3倍) 增加.
- 高密度发酵实现了HRP表达水平为200 mg/L,酶活性为1796 U/mL.
- 这与摇瓶发酵相比几乎增加了三倍,是报告中最高的.
结论:
- 同时表达HRP与特定的分子伴侣显著改善了Pichia pastoris的重组蛋白质生产.
- 优化的发酵策略,包括高密度种植,对于实现高产量的活性HRP至关重要.
- 开发的方法为工业规模的HRP生产提供了一个高效的平台.
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