Catalysis of an Essential Step in Vitamin B2 Biosynthesis by a Consortium of Broad Spectrum Hydrolases. Issue 17 (25th September 2015)

• Record Type: Journal Article
• Authors: Sarge, Sonja; Haase, Ilka; Illarionov, Boris; Laudert, Dietmar; Hohmann, Hans‐Peter; Bacher, Adelbert; Fischer, Markus
• Is Part Of: Chembiochem. Volume 16:Issue 17(2015)
• Page Start: 2466
• Page End: 2469
• Publication Date: 2015-09-25
• Subjects: biosynthesis -- haloacid dehalogenase -- hydrolases -- isoenzymes -- vitamin B2; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201500352
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: 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 of6 ( v max, 12 μmol mg −1 min −1 ; K M, 54 μm ) and of FMN ( v max, 25 μmol mg −1 min −1 ; K M, 135 μm ). A ycs E deletion mutant of B. subtilis was not riboflavin dependent. Two additional proteins (YwtE, YitU) that catalyse the hydrolysis of6 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. Abstract : The biosynthesis of riboflavin involves an essential dephosphorylation step. In Bacillus subtilis, used for the manufacture of the vitamin, this reaction can be catalysed by three different members of the haloacid dehalogenase (HAD) superfamily.