通过系统优化在大肠杆菌中高效生产cembratriene-ol。
Efficient production of cembratriene-ol in Escherichia coli via systematic optimization.
机构信息
The Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China.
The Key Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China.
出版信息
Microb Cell Fact. 2023 Jan 24;22(1):17. doi: 10.1186/s12934-023-02022-4.
BACKGROUND
The tobacco leaf-derived cembratriene-ol exhibits anti-insect effects, but its content in plants is scarce. Cembratriene-ol is difficult and inefficiently chemically synthesised due to its complex structure. Moreover, the titer of reported recombinant hosts producing cembratriene-ol was low and cannot be applied to industrial production.
RESULTS
In this study, Pantoea ananatis geranylgeranyl diphosphate synthase (CrtE) and Nicotiana tabacum cembratriene-ol synthase (CBTS) were heterologously expressed to synthsize the cembratriene-ol in Escherichia coli. Overexpression of cbts*, the 1-deoxy-D-xylulose 5-phosphate synthase gene dxs, and isopentenyl diphosphate isomerase gene idi promoted the production of cembratriene-ol. The cembratriene-ol titer was 1.53-folds higher than that of E. coli Z17 due to the systematic regulation of ggpps, cbts*, dxs, and idi expression. The production of cembratriene-ol was boosted via the overexpression of genes ispA, ispD, and ispF. The production level of cembratriene-ol in the optimal medium at 72 h was 8.55-folds higher than that before fermentation optimisation. The cembratriene-ol titer in the 15-L fermenter reached 371.2 mg L, which was the highest titer reported.
CONCLUSION
In this study, the production of cembratriene-ol in E. coli was significantly enhanced via systematic optimization. It was suggested that the recombinant E. coli producing cembratriene-ol constructed in this study has potential for industrial production and applications.
背景
来源于烟叶的吉马烯醇具有抗虫作用，但在植物中的含量稀少。由于其复杂的结构，吉马烯醇的化学合成既困难又低效。此外，报道的生产吉马烯醇的重组宿主的效价较低，无法应用于工业生产。
结果
在本研究中，异源表达了 Pantoea ananatis 香叶基香叶基二磷酸合酶（CrtE）和 Nicotiana tabacum 吉马烯醇合酶（CBTS）以在大肠杆菌中合成吉马烯醇。过表达 cbts*、1-脱氧-D-木酮糖 5-磷酸合酶基因 dxs 和异戊烯二磷酸异构酶基因 idi 促进了吉马烯醇的产生。由于 ggpps、cbts*、dxs 和 idi 表达的系统调节，吉马烯醇的产量比大肠杆菌 Z17 高 1.53 倍。通过过表达基因 ispA、ispD 和 ispF，吉马烯醇的产量得到了提高。在优化前发酵的最佳培养基中，72 小时后吉马烯醇的产量提高了 8.55 倍。在 15-L 发酵罐中，吉马烯醇的产量达到 371.2mg/L，这是报道的最高产量。
结论
本研究通过系统优化显著提高了大肠杆菌中吉马烯醇的产量。建议构建的生产吉马烯醇的重组大肠杆菌具有工业生产和应用的潜力。
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