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Catalysis of an Essential Step in Vitamin B 2 Biosynthesis by a Consortium of Broad Spectrum Hydrolases
Author(s) -
Sarge Sonja,
Haase Ilka,
Illarionov Boris,
Laudert Dietmar,
Hohmann HansPeter,
Bacher Adelbert,
Fischer Markus
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500352
Subject(s) - bacillus subtilis , riboflavin , biochemistry , biosynthesis , escherichia coli , cofactor , enzyme , chemistry , hydrolysis , dehalogenase , mutant , recombinant dna , biology , bacteria , gene , genetics
An enzyme catalysing the essential dephosphorylation of the riboflavin precursor, 5‐amino‐6‐ribitylamino‐2,4(1 H ,3 H )‐pyrimidinedione 5′‐phosphate ( 6 ), was purified about 800‐fold from a riboflavin‐producing Bacillus subtilis strain, and was assigned as the translation product of the ycs E gene by mass spectrometry. YcsE is a member of the large haloacid dehalogenase (HAD) superfamily. The recombinant protein was expressed in Escherichia coli . It catalyses the hydrolysis of 6 ( v max , 12 μmol mg −1 min −1 ; K M , 54 μ m ) and of FMN ( v max , 25 μmol mg −1 min −1 ; K M , 135 μ m ). A ycs E deletion mutant of B. subtilis was not riboflavin dependent. Two additional proteins (YwtE, YitU) that catalyse the hydrolysis of 6 at appreciable rates were identified by screening 13 putative HAD superfamily members from B. subtilis . The evolutionary processes that have resulted in the handling of an essential step in the biosynthesis of an essential cofactor by a consortium of promiscuous enzymes require further analysis.