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哈洛马纳斯坎帕尼尼斯菌株XH26的代谢工程,以消除竞争的途径,以增强生态素的生产
Zhiwan Shu1, Xin Zhang1, Rong Wang1
1Research Center of Basic Medical Science, Medical College of Qinghai University, Xining, 810016, China.
Scientific reports
|June 15, 2023
概括
阻断H.campaniensis*XH26中的一种代谢途径显著增加了ectoine的产生. 这种基因改造减少了甘氨酸副产品的形成,提高了有价值的护剂ectoine的产量.
科学领域:
- 微生物生物技术 微生物生物技术
- 代谢工程是代谢工程.
- 生物化学 生化学
背景情况:
- 埃克托因是一种高价值的 osmoprotectant,在市场上有很大的需求.
- 代谢途径可以限制像ectoine这样的商业重要化合物的产量.
- L-酸-4-半化物是ectoine合成的关键前体,但可以转向其他产品.
研究的目的:
- 为了提高 *Halomonas campaniensis* XH26.6. 在 *Halomonas campaniensis* XH26.6 中的生长率.
- 为了研究 homoserine dehydrogenase (hom) 基因在ectoine生物合成中的作用.
- 使用基因工程来阻断L-阿斯巴酸-4-半化物的代谢分流途径.
主要方法:
- 使用CRISPR/Cas9技术,无地淘汰同源基因.
- 在优化条件下在摇瓶和发酵器中培养工程和野生型菌株.
- 使用已确定的测量技术量化乙和贝产量.
主要成果:
- 与野生类型相比, *hom* 淘汰菌株 (XH26/Δhom) 呈现出明显增加的赤素产量 (351.13 mg (g CDW) -1 在摇瓶中,587.09 mg (g CDW) -1 在发酵器中) 相比 (239.18 mg (g CDW) -1 和385.03 mg (g CDW) -1).
- 与野生类型 (69.58 mg (g CDW) -1) 相比,工程菌株 (19.98 mg (g CDW) -1) 的贝他因合成减少了,这表明代谢转变.
- 阻断代谢分流通路有效地将前体基质重定向到ectoine生产.
结论:
- 针对性地破坏荷马氨酸脱酶通路是一种有效的策略,可以增加H. campaniensis*的ectoine产量.
- 减少贝他因合成似乎与增强的ectoine积累有关.
- 这种代谢工程方法有望改善工业规模的生化素生产.
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