Abstract
The thiO gene of Bacillus subtilis encodes an FAD-dependent glycine oxidase. This enzyme is a homotetramer with a monomer molecular mass of 42 kDa. In this paper, we demonstrate that ThiO is required for the biosynthesis of the thiazole moiety of thiamin pyrophosphate and describe the structure of the enzyme with N-acetylglycine bound at the active site. The closest structural relatives of ThiO are sarcosine oxidase and d-amino acid oxidase. The ThiO structure, as well as the observation that N-cyclopropylglycine is a good substrate, supports a hydride transfer mechanism for the enzyme. A mechanistic proposal for the role of ThiO in thiazole biosynthesis is also described.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Oxidoreductases / chemistry*
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Amino Acid Oxidoreductases / deficiency
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Amino Acid Oxidoreductases / genetics
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Amino Acid Oxidoreductases / physiology*
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Bacillus subtilis / enzymology*
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Bacillus subtilis / genetics
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Bacterial Proteins / physiology*
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Binding Sites
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Crystallography, X-Ray
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Deuterium / chemistry
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Flavin-Adenine Dinucleotide / chemistry
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Flavins / chemistry
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Flavoproteins / chemistry
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Flavoproteins / genetics
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Flavoproteins / metabolism
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Kinetics
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Mutation
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Nuclear Magnetic Resonance, Biomolecular
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Oxidation-Reduction
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Protein Structure, Quaternary
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Protein Structure, Tertiary
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Stereoisomerism
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Substrate Specificity
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Thiamine / biosynthesis*
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Thiazoles / chemistry
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Thiazoles / metabolism
Substances
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Bacterial Proteins
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Flavins
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Flavoproteins
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Thiazoles
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Flavin-Adenine Dinucleotide
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Deuterium
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Amino Acid Oxidoreductases
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glycine oxidase
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Thiamine