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Michael-type cyclizations in lantibiotic biosynthesis are reversible.

ABSTRACT: Lanthipeptides are members of the ribosomally synthesized and post-translationally modified peptides (RiPPs). They are generated in two biosynthetic steps: dehydration of Ser and Thr residues to the corresponding dehydroamino acids and subsequent conjugate addition by the thiol of Cys residues to generate the characteristic lanthionine and methyllanthionine thioether-bridged structures. Typically, a lanthipeptide contains multiple thioether cross-links. Recent studies have proposed that the final ring topology may be under thermodynamic control. If so, then the Michael-type cyclization reaction would need to be reversible, but such reversibility has never been demonstrated. We show here for the class I lanthipeptide cyclase NisC and class II lanthipeptide synthetase HalM2 that, indeed, the conjugate addition reactions are reversible and that the enzymes can open up all thioether rings in their products. We also propose that a His residue that is conserved among the lanthipeptide cyclases acts as the acid or base that protonates or generates the enolate intermediate during thioether ring formation and opening, respectively.

SUBMITTER: Yang X

PROVIDER: S-EPMC4433588 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature

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Michael-type cyclizations in lantibiotic biosynthesis are reversible.

Yang Xiao X van der Donk Wilfred A WA

ACS chemical biology 20150310 5

Lanthipeptides are members of the ribosomally synthesized and post-translationally modified peptides (RiPPs). They are generated in two biosynthetic steps: dehydration of Ser and Thr residues to the corresponding dehydroamino acids and subsequent conjugate addition by the thiol of Cys residues to generate the characteristic lanthionine and methyllanthionine thioether-bridged structures. Typically, a lanthipeptide contains multiple thioether cross-links. Recent studies have proposed that the fina ...[more]

PMID: 25723375

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Project description:Owing to extensive metagenomic studies, we now have access to numerous sequences of novel bacteriocin-like antimicrobial peptides encoded by various cultivable and noncultivable bacteria. However, relatively rarely, we even have access to these cultivable strains to examine the potency and the targets of the predicted bacteriocins. In this study, we evaluated a heterologous biosynthetic system to produce biologically active nonnative novel lantibiotics, which are modified bacteriocins. We chose Streptococcus mutans, a dental pathogen, as the host organism because it is genetically easy to manipulate and is inherently a prolific producer of various bacteriocins. We chose the S. mutans T8 strain as the host, which produces the lantibiotic mutacin II, to express 10 selected homologs of mutacin II identified from GenBank. These lantibiotic peptides either are novel or have been studied very minimally. The core regions of the selected lantibiotic peptides were fused to the leader sequence of the mutacin II peptide and integrated into the chromosome such that the core region of the native mutacin II was replaced with the new core sequences. By this approach, using the mutacin II biosynthesis machinery, we obtained one bioactive novel lantibiotic peptide with 52% different residues compared to the mutacin II core region. This unknown lantibiotic is encoded by Streptococcus agalactiae and Streptococcus ovuberis strains. Since this peptide displays some homology with nukacin ISK-1, we named it nukacin Spp. 2. This study demonstrated that the mutacin II biosynthesis machinery can be successfully used as an efficient system for the production of biologically active novel lantibiotics. IMPORTANCE In this study, we report for the first time that Streptococcus mutans can be used as a host to produce various nonnative lantibiotics. We showed that in the T8 strain, we could produce bioactive lacticin 481 and nukacin ISK-1, both of which are homologs of mutacin II, using T8's modification and secretion apparatus. Similarly, we also synthesized a novel bioactive lantibiotic, which we named nukacin Spp. 2.

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Michael-type cyclizations in lantibiotic biosynthesis are reversible.

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Michael-type cyclizations in lantibiotic biosynthesis are reversible.

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