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, 410081, China.
Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (MOE of China), National & Local Joint Engineering Laboratory for New Petro-Chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, 410081, China.
Microb Cell Fact. 2022 May 14;21(1):83. doi: 10.1186/s12934-022-01808-2.
Butenyl-spinosyn produced by Saccharopolyspora pogona exhibits strong insecticidal activity and a broad pesticidal spectrum. Currently, important functional genes involve in butenyl-spinosyn biosynthesis remain unknown, which leads to difficulty in efficiently understanding its regulatory mechanism, and improving its production by metabolic engineering.
Here, we identified a TetR family transcriptional regulator, SP_2854, that can positively regulate butenyl-spinosyn biosynthesis and affect strain growth, glucose consumption, and mycelial morphology in S. pogona. Using targeted metabolomic analyses, we found that SP_2854 overexpression enhanced glucose metabolism, while SP_2854 deletion had the opposite effect. To decipher the overproduction mechanism in detail, comparative proteomic analysis was carried out in the SP-2854 overexpressing mutant and the original strain, and we found that SP_2854 overexpression promoted the expression of proteins involved in glucose metabolism.
Our findings suggest that SP_2854 can affect strain growth and development and butenyl-spinosyn biosynthesis in S. pogona by controlling glucose metabolism. The strategy reported here will be valuable in paving the way for genetic engineering of regulatory elements in actinomycetes to improve important natural products production.
多杀菌素由 pogona 放线菌产生，具有很强的杀虫活性和广泛的杀虫谱。目前，涉及butenyl-spinosyn 生物合成的重要功能基因仍然未知，这导致难以有效地理解其调控机制，并通过代谢工程提高其产量。
在这里，我们鉴定了一个 TetR 家族转录调节因子 SP_2854，它可以正向调控 butenyl-spinosyn 生物合成，并影响 pogona 放线菌的生长、葡萄糖消耗和菌丝形态。通过靶向代谢组学分析，我们发现 SP_2854 过表达增强了葡萄糖代谢，而 SP_2854 缺失则有相反的效果。为了详细阐明过度生产的机制，我们对 SP-2854 过表达突变体和原始菌株进行了比较蛋白质组学分析，发现 SP_2854 过表达促进了参与葡萄糖代谢的蛋白质的表达。
我们的研究结果表明，SP_2854 可以通过控制葡萄糖代谢来影响 pogona 放线菌的生长和发育以及 butenyl-spinosyn 的生物合成。这里报道的策略将有助于放线菌调控元件的遗传工程，以提高重要天然产物的产量。
