Anhui Province Key Laboratory of Pollutant Sensitive Materials and Environmental Remediation, Huaibei Normal University, Huaibei, China.
Yota Bio-Engineering Co., Ltd, Rizhao, China.
Appl Microbiol Biotechnol. 2023 Feb;107(2-3):881-896. doi: 10.1007/s00253-022-12343-w. Epub 2022 Dec 31.
Epsilon-poly-L-lysine (ε-PL) is produced by Streptomyces species in acidic and aerobic conditions, which inevitably induces rapid generation of reactive oxygen species (ROS). The devastating effects of ROS on biomolecules and cell vitality have been well-studied, while the positive effects of ROS are rarely reported.
In this study, we found that a proper dose of intracellular ROS (about 3.3 μmol HO /g DCW) could induce a physiological modification to promote the ε-PL production (from 1.2 to 1.5 g/L). It resulted in larger sizes of colony and mycelial pellets as well as vibrant, aggregated, and more robust mycelia, which were of high capability of ROS detoxication. Physiological studies showed that appropriate doses of ROS activated the metabolism of the pentose phosphate pathway at both transcriptional and enzymatic levels, which was beneficial for biomass accumulation. The biosynthesis of lysine was also promoted in terms of transcriptional regulatory overexpression, increased transcription and enzymatic activity of key genes, larger pools of metabolites in the TCA cycle, replenishment pathway, and diaminoheptanedioic acid pathway. In addition, energy provision was ensured by activated metabolism of the TCA cycle, a larger pool of NADH, and higher activity of the electron transport system. Increased transcription of HrdD and pls further accelerated the ε-PL biosynthesis.
These results indicated that ROS at proper intracellular dose could act as an inducing signal to activate the ε-PL biosynthesis, which laid a foundation for further process regulation to maintain optimal ROS dose in industrial ε-PL production and was of theoretical and practical significance.
• A proper dose of intracellular ROS positively influences the ε-PL production. • Proper dose of ROS enhanced the mycelial activity and antioxidative capability. • ROS increased lysine synthesis metabolism, energy provision and pls expression.
ε-聚赖氨酸(ε-PL)是由链霉菌属在酸性和需氧条件下产生的,这不可避免地会导致活性氧(ROS)的快速产生。ROS 对生物分子和细胞活力的破坏性影响已经得到了很好的研究,而 ROS 的积极影响却很少被报道。
在这项研究中,我们发现适量的细胞内 ROS(约 3.3 μmol HO /g DCW)可以诱导生理修饰以促进 ε-PL 的产生(从 1.2 到 1.5 g/L)。这导致了较大的菌落和菌丝团大小以及充满活力、聚集和更健壮的菌丝,这些菌丝具有很强的 ROS 解毒能力。生理研究表明,适量的 ROS 激活了戊糖磷酸途径的代谢,在转录和酶水平上都得到了促进,这有利于生物量的积累。赖氨酸的生物合成也通过转录调控过表达、关键基因的转录和酶活性增加、TCA 循环、补充途径和二氨基庚二酸途径中的代谢物池增大得到促进。此外,通过 TCA 循环代谢的激活、更大的 NADH 池和电子传递系统更高的活性,为能量供应提供了保障。HrdD 和 pls 的转录增加进一步加速了 ε-PL 的生物合成。
这些结果表明,适量的细胞内 ROS 可以作为诱导信号来激活 ε-PL 的生物合成,这为进一步的过程调节奠定了基础,以维持工业 ε-PL 生产中的最佳 ROS 剂量,具有理论和实际意义。
• 适量的细胞内 ROS 对 ε-PL 的生产有积极影响。• 适量的 ROS 增强了菌丝的活性和抗氧化能力。• ROS 增加了赖氨酸合成代谢、能量供应和 pls 表达。