在Pichia pastoris中进行细分工程的氧体向序列的表征和工程
Cuifang Ye1, Haosen Hong1,2, Jucan Gao2
1Key Laboratory of Biomass Chemical Engineering of Ministry of Education, National Key Laboratory of Biobased Transportation Fuel Technology, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
Biotechnology and bioengineering
|April 3, 2024
概括
研究人员开发了一种高通量方法,以改善Pichia pastoris中的氧体向信号 (PTS1s). 这增强了代谢途径的工程,通过过氧化体细分化将单烯的产量提高了30%以上.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 细胞的分隔是细胞的分隔.
背景情况:
- 过氧体细分是代谢途径工程的一个强有力的策略.
- 皮奇亚牧羊人的过氧体具有可变的组成,限制了途径的复制.
- 缺乏特征化的过氧体向信号阻碍了在P. pastoris.中的应用.
研究的目的:
- 建立一种高通量选方法,用于评估P. pastoris.中的过氧体向信号 (PTS1) 效率.
- 为了识别用于增强氧体向的新型PTS1s.
- 将改进的PTS1用于P. pastoris.中的代谢途径工程.
主要方法:
- 开发了一种使用β-胡卜素生物合成途径的高通量选试验.
- 描述了25种内源性PTS1s,确定了3种高效的.
- 进行了定向进化来生成PTS1突变库,识别了51个新的PTS1s.
- 使用共聚焦显微镜验证了新的PTS1s.
- 使用已识别的PTS1s.重建过氧体中的geraniol生物合成途径.
主要成果:
- 确定了3种高效的内源性PTS1s.
- 发现了51个新的PTS1s (29个古典,22个非正典) 提高了准效率.
- 通过局部化geraniol通路到Peroxisomes,实现了单烯标位的30%以上的增加.
- 证明了增强的PTS1s对于过氧体代谢工程的实用性.
结论:
- 这项研究提供了一个扩展的工具包,用于P. pastoris.的过氧体细分.
- 新型PTS1s显著提高了Peroxisomes中的代谢途径复制的效率.
- 这项工作为使用P. pastoris peroxisomes的先进代谢工程应用奠定了基础.
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