Construction of the glycerol dehydratase (reuterin-producing) gene knockout mutant Lactobacillus reuteri and probiotics evaluation

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Lactobacillus reuteri is a heterofermentative lactic acid bacterium that naturally inhabits the gut of humans and other animals. The probiotic effects of L. reuterihave been proposed to be largely associated with the production of the broad-spectrum antimicrobial compound reuterin during anaerobic metabolism of glycerol. We determined the complete genome sequences of the reuterin-producing L. reuteri JCM 1112^T. L. reuteriJCM 1112^T has a unique cluster of 58 genes for the biosynthesis of reuterin and cobalamin (vitamin B_<12>). Two-dimensional nuclear magnetic resonance (2D-NMR) with ^<13>G_3-glycerol demonstrated that L. reuteri JCM 1112^T could convert glycerol to reuterin in vivo, substantiating the potential of L. reuteri JCM 1112^T to produce reuterin in the intestine. Given that glycerol is shown to be naturally present in feces, the acquired ability to produce reuterin and cobalamin is an adaptive evolutionary response that likely contributes to the probiotic properties of L. reuteri.