Abstract

Clostridium botulinum is an anaerobic sporeforming bacterium that is notorious for producing a potent neurotoxin. Spores of C. botulinum can survive mild food processing treatments and subsequently germinate, multiply, produce toxin and cause botulism. Control of spore germination and outgrowth is therefore essential for the safety of mildly processed foods. However, little is known about the process of spore germination in group II C. botulinum (gIICb), which are a major concern in chilled foods because they are psychrotrophic. The classical model of spore germination states that germination is triggered by the binding of a germinant molecule to a cognate germinant receptor. Remarkably, unlike many other sporeformers, gIICb has only one predicted canonical germinant receptor although it responds to multiple germinants. Therefore, we deleted the gerBAC locus that encodes this germinant receptor to determine its role in germination. Surprisingly, the deletion did not affect germination by any of the nutrient germinants, nor by the non-nutrient dodecylamine. We conclude that one or more other, so far unidentified, germinant receptors must be responsible for nutrient induced germination in gIICb. Furthermore, the gerBAC locus was strongly conserved with intact open reading frames in 159 gIICb genomes, suggesting that it has nevertheless an important function.

Highlights

  • Clostridium botulinum is an anaerobic sporeforming bacterium that is notorious for producing a potent neurotoxin

  • Human botulism is predominantly associated with group I and II C. botulinum, and these groups have been studied most extensively4–6. group II C. botulinum (gIICb) strains, in particular, are a major concern for the safety of mildly heat-processed refrigerated foods because they are psychrotrophic

  • Individual knockout of all ger operons in B. cereus ATCC 14579 failed to identify a role in germination for three (GerK, GerL and GerS) out of the seven Ger receptors, all operons were transcribed during sporulation[16]

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Summary

Introduction

Clostridium botulinum is an anaerobic sporeforming bacterium that is notorious for producing a potent neurotoxin. Spores of C. botulinum can survive mild food processing treatments and subsequently germinate, multiply, produce toxin and cause botulism. Spore germination is induced when specific nutrients bind to spore germinant receptors (GRs) in the spore membrane, signaling that conditions are favorable for outgrowth. This sets off a self-propagating and irreversible cascade of events starting with the release of monovalent cations H+, Na+, K+ and Ca2+-dipicolinic acid (Ca2+-DPA) from the spore core, followed by degradation of the spore cortex and core rehydration. Clarification of the structure-function relationship of Ger receptors has been hindered by the difficulty to isolate functional receptors, because they are membrane complexes composed of three subunits and associated with other proteins in a so-called germinosome complex[17,18]

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