Zhao Xueli, Lu Haisong, Peng Sen, Hang Haifeng, Aldahmash Waleed, Al-Ghadi Muath Q, Tang Weihua, Pei Jiang, Xun Wan, Guo Meijin, Mohsin Ali
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Shanghai Baying Bio Technology Co., Ltd., Shanghai, China.
Front Microbiol. 2025 Apr 23;16:1561042. doi: 10.3389/fmicb.2025.1561042. eCollection 2025.
Butenyl-spinosyn is a high-quality biological insecticide produced by that effectively targets a broad range of insect pests. However, the large-scale production of this insecticide is hindered by its low yield. Herein, based on prior comparative genomic analysis, five mutations were individually overexpressed in aG6. Subsequently, the combinatorial overexpression of (encoding NAD-glutamate dehydrogenase) and (encoding dTDP-glucose 4,6-dehydratase) in aG6 resulted in strain O1322-6746. The production of butenyl-spinosyn in O1322-6746 was 77.1% higher than that in aG6. Comparative targeted metabolomic analysis uncovered that O1322-6746 exhibited increased metabolic flux toward butenyl-spinosyn precursors. Furthermore, single-factor experiments, Plackett-Burman analysis and response surface methodology were performed to optimize the fermentation medium for O1322-6746. Ultimately, butenyl-spinosyn production was enhanced to 298.5 mg/L in a 5-L bioreactor, marking the highest yield ever reported. This work demonstrated that combining metabolic engineering with medium optimization is an effective strategy to improve butenyl-spinosyn production.
丁烯基多杀菌素是一种由[具体生产主体未给出]生产的优质生物杀虫剂，能有效针对多种害虫。然而，这种杀虫剂的大规模生产受到其低产量的阻碍。在此，基于先前的比较基因组分析，五个突变在aG6中分别过表达。随后，在aG6中[基因名称未完整给出，推测为编码NAD - 谷氨酸脱氢酶的基因]和[基因名称未完整给出，推测为编码dTDP - 葡萄糖4,6 - 脱水酶的基因]的组合过表达产生了菌株O1322 - 6746。O1322 - 6746中丁烯基多杀菌素的产量比aG6中的产量高77.1%。比较靶向代谢组学分析发现，O1322 - 6746对丁烯基多杀菌素前体的代谢通量增加。此外，进行了单因素实验、Plackett - Burman分析和响应面方法以优化O1322 - 6746的发酵培养基。最终，在5升生物反应器中丁烯基多杀菌素产量提高到298.5毫克/升，这是有报道以来的最高产量。这项工作表明，将代谢工程与培养基优化相结合是提高丁烯基多杀菌素产量的有效策略。
