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在共同养条件下,对缺乏AOX基因的Pichia pastoris中甲醇利用的转录基因洞察力

Xueyun Zheng¹, Zhifang Ye¹, Jiao Gao²

¹Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan, 430068, China.

Archives of microbiology

|May 9, 2025

概括

通过敲除酒精氧化酶基因和添加糖醇来提高蛋白质生产,Pichia pastoris的代谢工程. 这一战略通过减少有毒中间体,提高了使用可再生原料甲醇的生物制造.

科学领域:

生物技术是生物技术.
代谢工程是代谢工程.
合成生物学合成生物学

背景情况:

皮奇亚牧草是一种使用甲醇碳中性生物制造的关键平台.
在P. pastoris中,甲醇代谢产生有毒的甲,限制蛋白质生物合成.
代谢工程对于克服有效生物转化这些局限性至关重要.

研究的目的:

通过改造甲醇代谢来增强P. pastoris中异质蛋白质的产生.
为了减轻由有毒甲中间体引起的代谢压力.
提高生物制造效率,使用甲醇作为可再生原料.

主要方法:

通过对酒精氧化酶基因 (aox1和aox2) 的双重淘汰来实现代谢工程.
利用糖辅助基质补充剂来支持生长和蛋白质生产.
采用增强的绿色光蛋白 (EGFP) 作为量化模型异质产品.

主要成果:

与野生类型相比,工程化 ΔAOX1/2 菌株的生物质密度 (38.5 OD600) 和 EGFP 光 (494,723 个单位) 显著更高.
优化条件 (0.5%甲醇 + 0.4%甘油) 导致生物质增加32.8%,蛋白质生产率增加53.6%.
转录组分析表明通过协调代谢模块上调调节优化了甲醇利用率.

结论:

酒精氧化酶抑制与糖共同代谢相结合,有效降低甲醇衍生的代谢压力.
这一策略显著提高了P. pastoris.中异质蛋白的产量.
工程菌株为碳中和生物制造提供了一个更有效的平台.

关键词:

这就是Pichia pastoris.甲醇利用率的使用方法蛋白质生产蛋白质生产对转录组进行分析.

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