Hunan Provincial Key Laboratory for Microbial Molecular Biology, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, Hunan 410081, China.
Hunan Norchem Pharmaceutical Co., Ltd., Changsha, Hunan 410205, China.
J Biotechnol. 2024 Dec 10;396:127-139. doi: 10.1016/j.jbiotec.2024.10.010. Epub 2024 Nov 2.
Spinosad is an insecticide produced by Saccharopolyspora spinosa, and its larvicidal activity is considered a promising approach to combat crop pests. The aim of this study was to enhance the synthesis of spinosad through increasing the supply of acyl-CoAs precursor by the following steps. (i) Engineering the β-oxidation pathway by overexpressing key genes within the pathway to promote the synthesis of spinosad. The results showed that the overexpression of fadD, fadE, and fadA1 genes, as well as the co-expression of fadA1 and fadE genes, increased the yield of spinosad by 0.36-fold, 0.89-fold, 0.75-fold and 1.25-fold respectively. (ii) Employing combinatorial engineering of the β-oxidation pathway and ACC/PCC pathway to promote the synthesis of spinosad. The results showed that the co-expression of fadE and pccA, as well as accC and fadE, resulted in a 1.77-fold and 1.43-fold increase in spinosad production respectively. (iii) When exogenous triacylglycerol was added to the fermentation medium, the solely engineering of the β-oxidation pathway increased the yield of spinosad by 7.13-fold, reaching 427.23 mg/L. While the combinatorial engineering of both the β-oxidation pathway and ACC/PCC pathway increased the yield of spinosad by 9.61-fold, reaching 625.17 mg/L, and further optimization of the culture medium resulted in an even higher yield of spinosad, reaching 1293.43 mg/L. The results of this study indicate that the above combination strategy can promote the efficient biosynthesis of spinosad.
多杀菌素是由棘孢小单孢菌产生的杀虫剂，其杀虫活性被认为是防治作物害虫的一种很有前途的方法。本研究旨在通过以下步骤增加酰基辅酶 A 前体的供应来提高多杀菌素的合成。（i）通过过表达途径中的关键基因来工程化β-氧化途径，以促进多杀菌素的合成。结果表明，过表达 fadD、fadE 和 fadA1 基因以及 fadA1 和 fadE 基因的共表达分别将多杀菌素的产量提高了 0.36 倍、0.89 倍、0.75 倍和 1.25 倍。（ii）组合工程β-氧化途径和 ACC/PCC 途径以促进多杀菌素的合成。结果表明，fadE 和 pccA 的共表达以及 accC 和 fadE 的共表达分别使多杀菌素的产量增加了 1.77 倍和 1.43 倍。（iii）当向发酵培养基中添加外源性三酰基甘油时，单独工程化β-氧化途径将多杀菌素的产量提高了 7.13 倍，达到 427.23 mg/L。而β-氧化途径和 ACC/PCC 途径的组合工程化将多杀菌素的产量提高了 9.61 倍，达到 625.17 mg/L，进一步优化培养基进一步提高了多杀菌素的产量，达到 1293.43 mg/L。本研究结果表明，上述组合策略可以促进多杀菌素的高效生物合成。
