Evolution of vitamin B2 biosynthesis. A novel class of riboflavin synthase in Archaea

Journal of Molecular Biology
Markus FischerWolfgang Eisenreich

Abstract

The open reading frame MJ1184 of Methanococcus jannaschii with similarity to riboflavin synthase of Methanothermobacter thermoautotrophicus was cloned into an expression vector but was poorly expressed in an Escherichia coli host strain. However, a synthetic open reading frame that was optimized for expression in E.coli directed the synthesis of abundant amounts of a protein with an apparent subunit mass of 17.5 kDa. The protein was purified to apparent homogeneity. Hydrodynamic studies indicated a relative mass of 88 kDa suggesting a homopentamer structure. The enzyme was shown to catalyze the formation of riboflavin from 6,7-dimethyl-8-ribityllumazine at a rate of 24 nmol mg(-1) min(-1) at 40 degrees C. Divalent metal ions, preferably manganese or magnesium, are required for maximum activity. In contrast to pentameric archaeal type riboflavin synthases, orthologs from plants, fungi and eubacteria are trimeric proteins characterized by an internal sequence repeat with similar folding patterns. In these organisms the reaction is achieved by binding the two substrate molecules in an antiparallel orientation. With the enzyme of M.jannaschii, 13C NMR spectroscopy with 13C-labeled 6,7-dimethyl-8-ribityllumazine samples as substrate...Continue Reading

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Citations

Feb 3, 2006·Natural Product Reports·Tadhg P Begley
Jul 13, 2005·Natural Product Reports·Markus Fischer, Adelbert Bacher
Sep 28, 2007·Natural Product Reports·Duncan E ScottChris Abell
Nov 22, 2007·Photochemistry and Photobiology·Aba Losi
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Jul 27, 2005·Biochemical Society Transactions·M FischerA Bacher
Mar 16, 2011·Chembiochem : a European Journal of Chemical Biology·Markus Fischer, Adelbert Bacher
Jun 10, 2005·The Journal of Biological Chemistry·Boris IllarionovMarkus Fischer
Apr 10, 2021·International Journal of Biological Macromolecules·Gangming Xu, Suiqun Yang
Sep 1, 2010·EcoSal Plus·Markus Fischer, Adelbert Bacher
Feb 27, 2008·Archives of Biochemistry and Biophysics·Markus Fischer, Adelbert Bacher

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