College of Pharmacy, Binzhou Medical University, Yantai, 264003, People's Republic of China.
Microb Cell Fact. 2022 Oct 28;21(1):224. doi: 10.1186/s12934-022-01953-8.
ε-poly-L-lysine (ε-PL) is the main secondary metabolite of Streptomyces albulus, and it is widely used in the food industry. Polylysine synthetase (Pls) is the last enzyme in the ε-PL biosynthetic pathway. Our previous study revealed that Pls overexpressed in S. albulus CICC11022 result in the efficient production of ε-PL. In this study, a Pls gene knockout strain was initially constructed. Then, genomic, transcriptomic and metabolomic approaches were integrated to study the effects of the high expression and knockout of Pls on the gene expression and metabolite synthesis of S. albulus. The high expression of Pls resulted in 598 significantly differentially expressed genes (DEGs) and 425 known differential metabolites, whereas the inactivation of Pls resulted in 868 significant DEGs and 374 known differential metabolites. The expressions of 8 and 35 genes were negatively and positively associated with the Pls expression, respectively. Subsequently, the influence mechanism of the high expression and inactivation of Pls on the ε-PL biosynthetic pathway was elucidated. Twelve metabolites with 30% decreased yield in the high-expression strain of Pls but 30% increased production in the Pls knockout strain were identified. These results demonstrate the influence of Pls on the metabolism of S. albulus. The present work can provide the theoretical basis for improving the production capacity of ε-PL by means of metabolic engineering or developing bioactive substances derived from S. albulus.
ε-聚赖氨酸(ε-PL)是链霉菌属的主要次生代谢产物,广泛应用于食品工业。聚赖氨酸合成酶(Pls)是ε-PL 生物合成途径中的最后一种酶。我们之前的研究表明,在链霉菌属 CICC11022 中过表达 Pls 可导致 ε-PL 的高效生产。在本研究中,首先构建了 Pls 基因敲除菌株。然后,整合基因组、转录组和代谢组学方法,研究 Pls 的高表达和敲除对链霉菌属基因表达和代谢物合成的影响。Pls 的高表达导致 598 个显著差异表达基因(DEGs)和 425 个已知差异代谢物,而 Pls 的失活导致 868 个显著 DEGs 和 374 个已知差异代谢物。8 个和 35 个基因的表达与 Pls 表达呈负相关和正相关。随后,阐明了 Pls 的高表达和失活对 ε-PL 生物合成途径的影响机制。在 Pls 高表达菌株中,有 12 种代谢物的产量降低了 30%,而在 Pls 敲除菌株中,有 12 种代谢物的产量增加了 30%。这些结果表明 Pls 对链霉菌属的代谢有影响。本工作可为通过代谢工程提高 ε-PL 生产能力或开发链霉菌属来源的生物活性物质提供理论依据。