ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Microbiotechnology
This article is part of the Research TopicSynthetic Biology-Driven Microbial Synthesis of Bioactive Compounds and Industrial MaterialsView all articles
Tandem Solubility Enhancing Tags Enable the Heterologous Expression and in vitro Maturation of a Class IIa Bacteriocin, Clesteriocin A, Identified in Clostridium spp. CM038
Provisionally accepted
- 1University of Zurich, Zürich, Switzerland
- 2Universitat Zurich Institut fur Lebensmittelsicherheit und -hygiene, Zürich, Switzerland
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The Clostridium estertheticum complex (CEC) consists of closely related bacterial species that are mostly isolated from meat processing environment. Genome mining studies have recently established CEC as a source of class I bacteriocins. However, up until now, class II bacteriocins have not been reported from CEC. In the present study, we determined the presence of class II bacteriocin biosynthetic clusters in 33 CEC genomes through genome mining followed by bacteriocin production through heterologous expression in Escherichia coli. Six biosynthetic gene clusters belonging to class IIa (n = 1), IIb (n = 2), IId (n = 2) and an undefined class containing three precursor peptides were identified in six different CEC strains. Using molecular biology, we developed dedicated expression vectors for the class IIa bacteriocin, clesteriocin A. Its precursor peptide, CleA, and protease, CleB150, were initially expressed in insoluble form in E. coli. Through a systematic analysis of suitable solubility enhancing protein tags, both CleA and CleB150 were expressed in significant amounts of soluble fractions using tandem tags derived from Small Ubiquitin-like Modifier, superfolder Green fluorescent protein, Maltose binding protein and N-utilization substance. Mature clesteriocin A was obtained following in vitro maturation reactions between the tagged CleA and CleB150. The novel bacteriocin displayed antimicrobial activity against Listeria monocytogenes, which was mediated by the mptC gene. By combining genome mining and molecular biology, we have shown CEC is a source of novel class II bacteriocins that have potential for application as food biopreservatives.
Keywords: bacteriocin, Clostridium estertheticum complex, genome mining, Listeria monocytogenes, pediocin, Synthetic Biology
Received: 06 Dec 2025; Accepted: 22 Jan 2026.
Copyright: © 2026 Wambui, Tasara, Bigler and Stephan. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Joseph Wambui
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