在Pseudomonas putida中设计混合糖代谢道以产生香草酸
Peihan Wu1, Haifeng Ding1, Zhiqing Xu1
1School of Chemistry and Chemical Engineering/ State Key Laboratory Incubation Base for Green Processing of Chemical Engineering, Shihezi, 832003, PR China.
Synthetic and systems biotechnology
|October 27, 2025
概括
这项研究设计了*Pseudomonas putida*以高效地将基纤维素的混合糖转化为香草酸. 这种工程菌株克服了糖利用障碍,使得这种有价值的芳香化合物的高成本有效生物生产成为可能.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 生物技术是生物技术.
背景情况:
- 纤维素生物质水解产生混合糖 (葡萄糖,西洛斯,阿拉比诺斯),为微生物生物合成提供成本效益高的碳来源.
- 混合糖的协同利用受到碳催化剂抑制和代谢途径限制的阻碍,降低了整体碳利用效率.
- 香酸是一种有价值的芳香化合物,具有多种工业应用.
研究的目的:
- 设计*Pseudomonas putida*以协同利用混合糖 (葡萄糖,西洛糖,阿拉比诺糖) 来生产香草酸.
- 在混合糖利用中克服代谢瓶和碳催化剂抑制.
- 建立一个微生物平台,用于将纤维素酸化物转化为香草酸.
主要方法:
- 对*Pseudomonas putida*进行基因工程,以建立香草酸生物合成途径 (表达*OMT*,删除*vanAB*).
- 引入糖异构酶途径用于糖代谢.
- 代谢工程策略,包括删除*gcd*和*hexR*,以及过度表达的转基因酶和转酶,以平衡葡萄糖和西洛斯代谢.
- 组装了阿拉伯糖氧化途径,以完成三糖代谢.
主要成果:
- 工程 *P. putida* 菌株成功地从葡萄糖和树脂糖中积累了香草酸.
- 删除*gcd*和*hexR*,加上增强的酸通路酶,平衡葡萄糖-糖代谢.
- 最后的工程菌株 (VA12) 实现了葡萄糖,西洛斯和阿拉比诺斯的协同利用.
- 菌株VA12在料批发发酵中产生了2.75g/L的 vanilic 酸,使用葡萄糖,西洛糖和阿拉比诺糖的混合物.
结论:
- 这项研究成功地减少了碳催化剂抑制,用于协同利用混合糖.
- 这项工作代表了利用葡萄糖-西洛斯-阿拉比诺斯用于酸生产的首次演示.
- 工程 *P. putida* 平台展示了将纤维素化酸转化为有价值的化学产品的潜力.
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