Affiliation:
1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
Abstract
ε-Poly-L-lysine (ε-PL) is a natural and safe food preservative mainly produced by the aerobic, filamentous bacterium Streptomyces albulus. Therefore, it is crucial to breed superior ε-PL-producing strains to enhance fermentation efficiency to reduce production costs. Metabolic engineering is an effective measure for strain modification, but there are few reports on key genes for ε-PL biosynthesis. In this study, metabolic flux analysis was employed to identify potential key genes in ε-PL biosynthesis in S. albulus WG-608. A total of six potential key genes were identified. Three effective key genes (ppc, pyc and pls) were identified for the first time in ε-PL biosynthesis through overexpression experiments. It also presents the first demonstration of the promoting effects of ppc and pyc on ε-PL biosynthesis. Three genes were then co-expressed in S. albulus WG-608 to obtain OE-ppc-pyc-pls, which exhibited an 11.4% increase in ε-PL production compared to S. albulus WG-608, with a 25.5% increase in specific ε-PL production. Finally, the metabolic flux analysis of OE-ppc-pyc-pls compared to S. albulus WG-608 demonstrated that OE-ppc-pyc-pls successfully altered the metabolic flux as expected. This study not only provides a theoretical basis for the metabolic engineering of ε-PL-producing strains but also provides an effective approach for the metabolic engineering of other metabolites.
Funder
Key R&D Program of Jiangsu Province
National Key R&D Program of China
Evaluation Results of 2022 Open Subjects of Key Experiment of Ministry of Education in Industrial Biotechnology
Subject
Plant Science,Biochemistry, Genetics and Molecular Biology (miscellaneous),Food Science