Bacillus megaterium has both a functional BluB protein required for DMB synthesis and a related flavoprotein that forms a stable radical species.
Bacillus megaterium has both a functional BluB protein required for DMB synthesis and a related flavoprotein that forms a stable radical species.
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DOI：
10.1371/journal.pone.0055708
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lawrence AD
中科院分区：
文献类型：
--
作者：
Collins HF;Biedendieck R;Leech HK;Gray M;Escalante-Semerena JC;McLean KJ;Munro AW;Rigby SE;Warren MJ;Lawrence AD
Despite the extensive study of the biosynthesis of the complex molecule B12 (cobalamin), the mechanism by which the lower ligand 5,6-dimethylbenzimidazole (DMB) is formed has remained something of a mystery. However, recent work has identified and characterized a DMB-synthase (BluB) responsible for the oxygen-dependent, single enzyme conversion of FMN to DMB. In this work, we have identified BluB homologs from the aerobic purple, nonsulfur, photosynthetic bacterium Rhodobacter capsulatus and the aerobic soil bacterium Bacillus megaterium and have demonstrated DMB synthesis by the use of a novel complementation assay in which a B12 deficient strain, substituted with the precursor cobinamide is recovered either by the addition of DMB or by the recombinant expression of a bluB gene. The DMB-synthetic activity of the purified recombinant BluB enzymes was further confirmed in vitro by providing the enzyme with FMNH2 and oxygen and observing the formation of DMB by HPLC. The formation of a 4a-peroxyflavin intermediate, the first step in the oxygen dependent mechanism of DMB biosynthesis, is reported here and is the first intermediate in the enzyme catalysed reaction to be demonstrated experimentally to date. The identification and characterization of an FMN-binding protein found on the cobI operon of B. megaterium, CbiY, is also detailed, revealing an FMN-containing enzyme which is able to stabilize a blue flavin semiquinone upon reduction with a 1-electron donor.
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影响因子：
3.2
作者：
Anderson, Peter J.;Lango, Jozsef;Roth, John R.
通讯作者：
Roth, John R.
影响因子：
2
作者：
Kay, CWM;Schleicher, E;Weber, S
通讯作者：
Weber, S
影响因子：
5.2
作者：
Maggio-Hall, LA;Dorrestein, PC;Begley, TP
通讯作者：
Begley, TP
DOI：
10.1073/pnas.0609270104
发表时间：
2007-02-20
影响因子：
11.1
作者：
Gray, Michael J.;Escalante-Semerena, Jorge C.
通讯作者：
Escalante-Semerena, Jorge C.
影响因子：
2.9
作者：
MASSEY, V;PALMER, G
通讯作者：
PALMER, G