Supplementary Material for: Improvement of Spinosad Production upon Utilization of Oils and Manipulation of β-Oxidation in a High-Producing <b><i>Saccharopolyspora spinosa</i></b> Strain

Spinosad, a member of polyketide-derived macrolides produced in the actinomycete <i>Saccharopolyspora spinosa</i>, has been developed as a broad-spectrum and effective insecticide. The β-oxidation pathway could be an important source of building blocks for the biosynthesis of spinosad, thus the effect of vegetable oils on the production of spinosad in a high-yield strain was investigated. The spinosad production increased significantly with the addition of strawberry seed oil (511.64 mg/L) and camellia oil (520.07 mg/L) compared to the control group without oil (285.76 mg/L) and soybean oil group (398.11 mg/L). It also revealed that the addition of oils would affect the expression of genes involved in fatty acid metabolism, precursor supply, and oxidative stress. The genetically engineered strain, in which <i>fadD1</i> and <i>fadE</i> genes of <i>Streptomyces coelicolor</i> were inserted, produced spinosad up to 784.72 mg/L in the medium containing camellia oil, while a higher spinosad production level (843.40 mg/L) was detected with the addition of 0.01 mM of thiourea.

多杀菌素（Spinosad）是由放线菌糖多孢红霉菌（Saccharopolyspora spinosa）合成的聚酮类大环内酯化合物，已被开发为广谱高效杀虫剂。β-氧化途径（β-oxidation pathway）可能是多杀菌素生物合成的重要前体来源，因此本研究考察了植物油对高产菌株多杀菌素合成产量的影响。实验结果显示，与无油对照组（285.76 mg/L）及大豆油组（398.11 mg/L）相比，添加草莓籽油（511.64 mg/L）与茶籽油（520.07 mg/L）可显著提升多杀菌素产量。研究同时发现，油脂添加会影响脂肪酸代谢、前体供给及氧化应激相关基因的表达。通过插入天蓝色链霉菌（Streptomyces coelicolor）fadD1与fadE基因构建的基因工程菌株，在添加茶籽油的培养基中多杀菌素产量可达784.72 mg/L；而当添加0.01 mM硫脲时，多杀菌素产量进一步提升至843.40 mg/L。