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通过Kozak优化的2A多基因表达系统,增强Penicillium brevicompactum中的菌酸生产

Jishou Wu¹, TingTing Hu¹, Lin Lin²

¹State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, People's Republic of China.

|April 13, 2025

概括

我们优化了Penicillium brevicompactum中的基因表达,使用Kozak序列的2A系统. 这增加了49%的mycophenolic acid (MPA) 生产,并改善了真菌生长,为代谢工程提供了新的工具.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
工业微生物学工业微生物学

背景情况:

虫 (Penicillium brevicompactum) 对于工业菌酸 (MPA) 生产至关重要,但其代谢工程和基因调节尚未得到充分探索.
不有效的多基因表达策略阻碍了生物合成途径的优化,导致产量减少和细胞生长受损.
目前在P. brevicompactum中协调基因表达的方法需要优化以提高产品合成.

研究的目的:

通过结合Kozak序列,优化P. brevicompactum中的2A多基因表达系统.
增强关键的氨酸通路基因 (ERG10和ERG13) 的转录,以增加前体的可用性.
为了改善mycophenolic acid (MPA) 生产,并减轻代谢工程菌株的生长抑制.

主要方法:

在P. brevicompactum.中的2A多基因表达系统中引入Kozak序列.
使用RT-qPCR量化ERG10和ERG13基因转录水平.
与野生类型相比,在工程菌株中测量基酸 (MPA) 产量和菌株干重.

主要成果:

与PP-1013菌株相比,具有Kozak序列的PP-K10K13菌株显示ERG10的77.02%增加和ERG13转录的67.63%增加.
改造品种的酸 (MPA) 产量达到4.30g/L,比野生品种提高49.31%.
工程菌株表现出增强的生长,由增加的细胞干重量表明,这表明代谢负担减少.

结论:

优化的2A表达系统与Kozak序列有效地增强MPA生物合成,并恢复P. brevicompactum的正常生长.
这种改进的系统是多基因表达和P. brevicompactum. 未来遗传修饰的宝贵工具.
这项研究提出了一种新的策略,用于在状真菌中开发高效的2A多基因表达系统.

关键词:

2A类的.科扎克序列是什么意思多基因表达的多基因表达基酸是什么基酸是什么虫虫 (Penicillium brevicompactum) 是一种虫,它可以发育在任何地方.

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