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Volume 149, Issue 4

-5,6-Dimethylbenzimidazole adenine dinucleotide (-DAD), a putative new intermediate of coenzyme B biosynthesis in Free

Abstract

The CobT enzyme of was shown to have NAD-dependent ADPribosyltransferase activity. The CobT enzyme transferred the ADPribosyl moiety of NAD onto 5,6-dimethylbenzimidazole (DMB) yielding a new dinucleotide, namely -5,6-dimethylbenzimidazole adenine dinucleotide (-DAD), whose identity was established by mass spectrometry. The -(--ribosyl)-5,6-dimethylbenzimidazoyl moiety (-ribazole) of -DAD was incorporated into adenosylcobalamin (AdoCbl) by cell-free extracts of , indicating that -DAD served as an intermediate of AdoCbl biosynthesis. The rate of transfer of the ADPribosyl moiety was slower than the rate of transfer of the phosphoribosyl moiety of nicotinate mononucleotide (NaMN) to DMB. The CobT enzyme displayed a low for NaMN (0·51 mM) relative to the one for NAD (9 mM); nicotinate adenine dinucleotide (NaAD) and nicotinamide mononucleotide (NMN) also served as substrates for CobT. In spite of the high of CobT for NAD, the latter is proposed to be a relevant physiological substrate of CobT, given that the intracellular concentrations of NaMN, NMN and NaAD in actively growing are undetectable. Evidence shows that extracts of contain an as-yet unidentified dinucleotide pyrophosphatase that can cleave -DAD into -ribazole-5′-P and AMP; -ribazole-5′-P can then enter the AdoCbl biosynthetic pathway.

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Keyword(s): -P, phosphate , a.m.u., atomic mass unit , AdoCbi, adenosylcobinamide , AdoCbl, adenosylcobalamin , DMB, 5,6-dimethylbenzimidazole , ESIMS, electrospray ionization mass spectrometry , NaAD, nicotinate adenine dinucleotide , NaMN, nicotinate mononucleotide , NLA, nucleotide loop assembly , NMN, nicotinamide mononucleotide , RP-HPLC, reverse-phase HPLC , α-DAD, α-5,6-dimethylbenzimidazole adenine dinucleotide and α-ribazole, N1-(α-d-ribosyl)-5,6-dimethylbenzimidazole
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2003-04-01
2024-04-25
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α-5,6-Dimethylbenzimidazole adenine dinucleotide (α-DAD), a putative new intermediate of coenzyme B12 biosynthesis in Salmonella typhimurium
Microbiology 149, 983 (2003); https://doi.org/10.1099/mic.0.26040-0
/content/journal/micro/10.1099/mic.0.26040-0
/content/journal/micro/10.1099/mic.0.26040-0
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