Hunan Provincial Key Laboratory for Microbial Molecular Biology, State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, China.
Hunan Provincial Key Laboratory for Microbial Molecular Biology, State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, China.
Gene. 2021 Jan 15;766:145130. doi: 10.1016/j.gene.2020.145130. Epub 2020 Sep 7.
The LytTR family two-component system widely exists in bacterial cells and plays an important role in metabolic regulation. The lytS-L gene that encodes for a LytTR family sensor kinase was knocked out to study its influence on the growth, phenotype, and the biosynthesis of the insecticidal polyketide butenyl-spinosyn in Saccharopolyspora pogona NRRL 30141 (S. pogona). High performance liquid chromatography (HPLC) results showed that the butenyl-spinosyn yield of the lytS-L knockout mutant decreased by 58.9% compared with that of the parental strain. This is manifested by a weak toxicity of the mutant against the insect Helicoverpa assulta (H. armigera). Comparative proteomic analysis revealed the expression characteristics of the proteins in S. pogona and S. pogona-ΔlytS-L: a total of 14 proteins involved in energy metabolism were down-regulated, 9 proteins related to carbon metabolism such as glycolysis, and tricarboxylic acid cycle (TCA) were up-regulated, while 13 proteins involved in the biosynthesis of butenyl-spinosyn were down-regulated (fold change >1.2 or< 0.83). The qRT-PCR (Quantitative Real-time PCR) analysis illustrated that the changes in the expression levels of transcription and translation of the identified genes were consistent. This study explores the function of the two-component system of the LytTR family in S. pogona and shows that the lytS-L gene has an important influence on regulating primary metabolism and butenyl-spinosyn biosynthesis of S. pogona.
LytTR 家族双组份系统广泛存在于细菌细胞中，在代谢调控中发挥着重要作用。敲除编码 LytTR 家族传感器激酶的 lytS-L 基因，以研究其对昆虫杀虫聚酮butenyl-spinosyn 在昆虫病原放线菌 S. pogona NRRL 30141（S. pogona）中生长、表型和生物合成的影响。高效液相色谱（HPLC）结果表明，与亲本菌株相比，lytS-L 敲除突变体的 butenyl-spinosyn 产量下降了 58.9%。这表现为突变体对昆虫烟青虫（H. armigera）的毒性较弱。比较蛋白质组学分析揭示了 S. pogona 和 S. pogona-ΔlytS-L 中蛋白质的表达特征：共下调了 14 种参与能量代谢的蛋白质，上调了 9 种与糖酵解和三羧酸循环（TCA）等碳代谢相关的蛋白质，而与 butenyl-spinosyn 生物合成相关的 13 种蛋白质下调（倍数变化>1.2 或<0.83）。qRT-PCR（定量实时 PCR）分析表明，鉴定基因的转录和翻译表达水平的变化一致。本研究探讨了 LytTR 家族双组份系统在 S. pogona 中的功能，表明 lytS-L 基因对调节 S. pogona 的初级代谢和 butenyl-spinosyn 生物合成有重要影响。
