MtpB, a member of the MttB superfamily from the human intestinal acetogen Eubacterium limosum, catalyzes proline betaine demethylation

• Published in: The Journal of biological chemistry 2019-09, Vol.294 (37), p.13697-13707
• Main Authors: Picking, Jonathan W., Behrman, Edward J., Zhang, Liwen, Krzycki, Joseph A.
• Format: Article
• Language: English
• Subjects: acetogenesis; bacterial metabolism; Betaine - metabolism; cobalamin; Demethylation; Energy Metabolism; enzyme catalysis; Eubacterium - enzymology; Eubacterium - metabolism; folate; Folic Acid - metabolism; Humans; Intestines - microbiology; Metabolism; Methylamines - metabolism; Methylation; Methyltransferases - metabolism; microbiology; microbiome; Microbiota; one-carbon metabolism; Proline - analogs & derivatives; Proline - metabolism; Tetrahydrofolates - metabolism; trimethylamine methyltransferase
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA119.009886

The trimethylamine methyltransferase MttB is the founding member of a widely distributed superfamily of microbial proteins. Genes encoding most members of the MttB superfamily lack the codon for pyrrolysine that distinguishes previously characterized trimethylamine methyltransferases, leaving the function(s) of most of the enzymes in this superfamily unknown. Here, investigating the MttB family member MtpB from the human intestinal isolate Eubacterium limosum ATCC 8486, an acetogen that excretes N-methyl proline during growth on proline betaine, we demonstrate that MtpB catalyzes anoxic demethylation of proline betaine. MtpB along with MtqC (a corrinoid protein) and MtqA (a methylcorrinoid:tetrahydrofolate methyltransferase) was much more abundant in E. limosum cells grown on proline betaine than on lactate. We observed that recombinant MtpB methylates Co(I)-MtqC in the presence of proline betaine and that other quaternary amines are much less preferred substrates. MtpB, MtqC, and MtqA catalyze tetrahydrofolate methylation with proline betaine, thereby forming a key intermediate in the Wood–Ljungdahl acetogenesis pathway. To our knowledge, MtpB methylation of Co(I)-MtqC for the subsequent methylation of tetrahydrofolate represents the first described anoxic mechanism of proline betaine demethylation. The activities of MtpB and associated proteins in acetogens or other anaerobes provide a possible mechanism for the production of N-methyl proline by the gut microbiome. MtpB's activity characterized here strengthens the hypothesis that much of the MttB superfamily comprises quaternary amine-dependent methyltransferases.