Recombinant production of the lantibiotic nisin using Corynebacterium glutamicum in a two-step process

Microbial Cell Factories

Nadav S. Bar¹  Kirill V. Ovchinnikov²  Dzung B. Diep²  Dominik Weixler³  Christian U. Riedel³  Oliver Goldbeck³  Sebastian Reich³  Bernhard J. Eikmanns³  Max Berghoff³  Gerd M. Seibold⁴  Christoph Wittmann⁵ 

[1] Department of Chemical Engineering, Norwegian University of Science and Technology, Trondheim, Norway;Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås, Norway;Institute of Microbiology and Biotechnology, University of Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany;Institute of Microbiology and Biotechnology, University of Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany;Department of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark;Institute of Systems Biotechnology, Saarland University, Saarbrücken, Germany;

关键词: Corynebacterium glutamicum;    Bacteriocin;    Nisin;    Recombinant production;    Pre-nisin;    Biosensor;   

DOI  :  10.1186/s12934-022-01739-y

来源: Springer

【 摘 要 】

BackgroundThe bacteriocin nisin is naturally produced by Lactococcus lactis as an inactive prepeptide that is modified posttranslationally resulting in five (methyl-)lanthionine rings characteristic for class Ia bacteriocins. Export and proteolytic cleavage of the leader peptide results in release of active nisin. By targeting the universal peptidoglycan precursor lipid II, nisin has a broad target spectrum including important human pathogens such as Listeria monocytogenes and methicillin-resistant Staphylococcus aureus strains. Industrial nisin production is currently performed using natural producer strains resulting in rather low product purity and limiting its application to preservation of dairy food products.ResultsWe established heterologous nisin production using the biotechnological workhorse organism Corynebacterium glutamicum in a two-step process. We demonstrate successful biosynthesis and export of fully modified prenisin and its activation to mature nisin by a purified, soluble variant of the nisin protease NisP (sNisP) produced in Escherichia coli. Active nisin was detected by a L. lactis sensor strain with strictly nisin-dependent expression of the fluorescent protein mCherry. Following activation by sNisP, supernatants of the recombinant C. glutamicum producer strain cultivated in standard batch fermentations contained at least 1.25 mg/l active nisin.ConclusionsWe demonstrate successful implementation of a two-step process for recombinant production of active nisin with C. glutamicum. This extends the spectrum of bioactive compounds that may be produced using C. glutamicum to a bacteriocin harboring complex posttranslational modifications. Our results provide a basis for further studies to optimize product yields, transfer production to sustainable substrates and purification of pharmaceutical grade nisin.

【 授权许可】

CC BY