Beijing Key Laboratory of Nutrition and Health and Food Safety, Nutrition and Health Research Institute, COFCO, Beijing, 102209, People's Republic of China.
Qilu Pharmaceutical (Inner Mongolia) Co., Ltd., Hohhot, 010020, Inner Mongolia, People's Republic of China.
Sci Rep. 2021 Jul 20;11(1):14779. doi: 10.1038/s41598-021-94251-z.
Saccharopolyspora spinosa is a well-known actinomycete for producing the secondary metabolites, spinosad, which is a potent insecticides possessing both efficiency and safety. In the previous researches, great efforts, including physical mutagenesis, fermentation optimization, genetic manipulation and other methods, have been employed to increase the yield of spinosad to hundreds of folds from the low-yield strain. However, the metabolic network in S. spinosa still remained un-revealed. In this study, two S. spinosa strains with different spinosad production capability were fermented and sampled at three fermentation periods. Then the total RNA of these samples was isolated and sequenced to construct the transcriptome libraries. Through transcriptomic analysis, large numbers of differentially expressed genes were identified and classified according to their different functions. According to the results, spnI and spnP were suggested as the bottleneck during spinosad biosynthesis. Primary metabolic pathways such as carbon metabolic pathways exhibited close relationship with spinosad formation, as pyruvate and phosphoenolpyruvic acid were suggested to accumulate in spinosad high-yield strain during fermentation. The addition of soybean oil in the fermentation medium activated the lipid metabolism pathway, enhancing spinosad production. Glutamic acid and aspartic acid were suggested to be the most important amino acids and might participate in spinosad biosynthesis.
棘孢小单孢菌是一种生产具有高效和安全特点的杀虫剂——多杀菌素的著名放线菌。在以前的研究中,人们采用物理诱变、发酵优化、遗传操作等方法,使多杀菌素的产量从低产菌株提高了数百倍。然而,棘孢小单孢菌的代谢网络仍然没有被揭示。在这项研究中,我们对具有不同多杀菌素生产能力的两株棘孢小单孢菌进行发酵,并在三个发酵阶段取样。然后分离和测序这些样品的总 RNA,构建转录组文库。通过转录组分析,根据它们不同的功能,鉴定和分类了大量差异表达基因。结果表明,spnI 和 spnP 被认为是多杀菌素生物合成中的瓶颈。初级代谢途径,如碳代谢途径,与多杀菌素的形成密切相关,因为在发酵过程中,丙酮酸和磷酸烯醇式丙酮酸被认为在高产多杀菌素菌株中积累。在发酵培养基中添加大豆油激活了脂代谢途径,从而提高了多杀菌素的产量。谷氨酸和天冬氨酸被认为是最重要的氨基酸,可能参与多杀菌素的生物合成。