Physiology of γ-aminobutyric acid production by Akkermansia muciniphila

Gut bacteria hold the potential to produce a broad range of metabolites that can modulate human functions, including molecules with neuroactive potential. One such molecule is gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter of the central nervous system in animals. Metagenomic analyses suggest that the genomes of many gut bacteria encode glutamate decarboxylase (GAD), the enzyme that catalyzes GABA production. The genome of Akkermansia muciniphila, a mucin specialist and potential next-generation probiotic from the human gut, is predicted to encode GAD, suggesting a contributing role in GABA production in the human gut. In this study, A. muciniphila was grown in batch cultures with and without pH control. In both experiments, A. muciniphila was found to produce GABA as a response to acid (pH <5.5), although only when GABA precursors, either glutamate or glutamine, were present in the medium. Proteomic analysis comparing A. muciniphila grown with and without precursors at pH 4 did not show a difference in GAD expression, suggesting that it is expressed regardless of the presence of GABA precursors. To further investigate the function of A. muciniphila GAD, we heterologously expressed the gad gene (encoded by locus tag Amuc_0372) with a His tag in Escherichia coli and purified the GAD protein. Enzyme assays showed GAD activity in a pH range between 4 and 6, with the highest specific activity at pH 5 of 144 +/- 16 mu M GABA/min/mg. Overall, our results demonstrate the ability of A. muciniphila to produce GABA as an acid response and unravel the conditions under which GABA production in A. muciniphila occurs.

Automated summary

Gut bacteria have the ability to produce metabolites, including neuroactive molecules like GABA, that can modulate human functions. This study focuses on Akkermansia muciniphila, a potential probiotic, and its production of GABA. The study shows that A. muciniphila can produce GABA in response to acid, but only when GABA precursors, such as glutamate or glutamine, are present. The study also demonstrates that GAD, the enzyme responsible for GABA production, is expressed by A. muciniphila regardless of the presence of GABA precursors. Enzyme assays further reveal the specific conditions under which GABA production by A. muciniphila occurs.