Synthesis of a Novel Lantibiotic Using Mutacin II Biosynthesis Apparatus

Biswas, Saswati

doi:10.1128/spectrum.03030-22

Cited by 3 publications

(1 citation statement)

References 56 publications

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“…That is, we fused the leader peptide to the N -terminus of different other core peptides to create hybrid precursor peptides, which are likely modified by SboM. This strategy for generating natural and/or hybrid RiPP variants has been well documented in previous studies 79 , 80 . Second, SboT was co-expressed in UniBioCat reactions to mature lanthipeptides by removing the leader peptide.…”

Section: Resultsmentioning

confidence: 99%

Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides

Liu,

Ji,

et al. 2024

Nat Commun

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Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a major class of natural products with diverse chemical structures and potent biological activities. A vast majority of RiPP gene clusters remain unexplored in microbial genomes, which is partially due to the lack of rapid and efficient heterologous expression systems for RiPP characterization and biosynthesis. Here, we report a unified biocatalysis (UniBioCat) system based on cell-free gene expression for rapid biosynthesis and engineering of RiPPs. We demonstrate UniBioCat by reconstituting a full biosynthetic pathway for de novo biosynthesis of salivaricin B, a lanthipeptide RiPP. Next, we delete several protease/peptidase genes from the source strain to enhance the performance of UniBioCat, which then can synthesize and screen salivaricin B variants with enhanced antimicrobial activity. Finally, we show that UniBioCat is generalizable by synthesizing and evaluating the bioactivity of ten uncharacterized lanthipeptides. We expect UniBioCat to accelerate the discovery, characterization, and synthesis of RiPPs.

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Section: Resultsmentioning

confidence: 99%

Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides

Liu,

Ji,

et al. 2024

Nat Commun

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A review of current antibiotic resistance and promising antibiotics with novel modes of action to combat antibiotic resistance

Chen,

Kumar,

2023

Arch Microbiol

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Opposing genetic polymorphisms of two ABC transporters contribute to the variation of nukacin resistance in Streptococcus mutans

Sadaoka,

Le,

Kawada-Matsuo

et al. 2024

Appl Environ Microbiol

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Streptococcus mutans is a cariogenic bacterium that produces a variety of bacteriocins and retains resistance to these bacteriocins. In this study, we investigated the susceptibility of 127 S . mutans strains to nukacins produced by Staphylococcus spp., which are commensal bacteria in humans. We detected diverse susceptibilities among strains. Nineteen strains had a disrupted LctF (type I), which is responsible for nukacin susceptibility, whereas the remaining 108 strains had an intact LctF (type II) and displayed resistance to nukacins. However, the type I strains still showed resistance to nukacins to some extent. Interestingly, 18/19 (94.7%) type I strains carried a mukA-T locus, which is related to the synthesis of mutacin K8, and mukFEG , an ABC transporter. In contrast, among type II strains, only 6/108 strains (5.6%) had both the mukA-T locus and mukFEG , 19/108 strains (17.6%) carried only mukFEG , and 83/108 strains (76.9%) harbored neither mukA-T nor mukFEG . We also found that MukF had two variants: 305 amino acids (type α) and 302 amino acids (type β). All type I strains showed a type α (MukFα), whereas most type II strains with mukFEG (22/25 strains) had a type β (MukFβ). Then, we constructed a mukFEG -deletion mutant complemented with MukFαEG or MukFβEG and found that only MukFαEG was involved in nukacin resistance. The nukacin resistance capability of type II-LctFEG was stronger than that of MukFαEG. In conclusion, we identified a novel nukacin resistance factor, MukFEG, and either LctFEG or MukFEG was active in most strains via genetic polymorphisms depending on mukA-T genes. IMPORTANCE Streptococcus mutans is an important pathogenic bacterium not only for dental caries but also for systemic diseases. S. mutans is known to produce a variety of bacteriocins and to retain resistance these bacteriocins. In this study, two ABC transporters, LctFEG and MukFEG, were implicated in nukacin resistance and each ABC transporter has two subtypes, active and inactive. Of the two ABC transporters, only one ABC transporter was always resistant, while the other ABC transporter was inactivated by genetic mutation. Interestingly, this phenomenon was defined by the presence or absence of the mutacin K8 synthesis gene region, one of the bacteriocins of S. mutans . This suggests that the resistance acquisition is tightly controlled in each strain. This study provides important evidence that the insertion of bacteriocin synthesis genes is involved in the induction of genetic polymorphisms and suggests that bacteriocin synthesis genes may play an important role in bacterial evolution.

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Synthesis of a Novel Lantibiotic Using Mutacin II Biosynthesis Apparatus

Cited by 3 publications

References 56 publications

Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides

Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides

A review of current antibiotic resistance and promising antibiotics with novel modes of action to combat antibiotic resistance

Opposing genetic polymorphisms of two ABC transporters contribute to the variation of nukacin resistance in Streptococcus mutans

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