The dddP gene of Roseovarius nubinhibens encodes a novel lyase that cleaves dimethylsulfoniopropionate into acrylate plus dimethyl sulfide.
The cloned dddP gene of the marine bacterium Roseovarius nubinhibens allows Escherichia coli to form the volatile dimethyl sulfide (DMS) from dimethylsulfoniopropionate (DMSP), an abundant anti-stress compatible solute made by many marine plankton and macroalgae. Using purified DddP, we show here that this enzyme is a DMSP lyase that cleaves DMSP to DMS plus acrylate. DddP forms a functional homodimeric enzyme, has a pH optimum of 6.0 and was a K(m) of approximately 14 mM for the DMSP substrate. DddP belongs to the M24B family of peptidases, some members of which have metal cofactors. However, the metal chelators EDTA and bipyridyl did not affect DddP activity in vitro and the as-isolated enzyme did not contain metal ions. Thus, DddP resembles those members of the M24B family, such as creatinase, which also act on a non-peptide substrate and have no metal cofactor. Site-directed mutagenesis of the active-site region of DddP completely abolished its activity. Another enzyme, termed DddL, which occurs in other alphaproteobacteria, had also been shown to generate DMS plus acrylate from DMSP. However, DddL and DddP have no sequence similarity to each other, so DddP represents a second, wholly different class of DMSP lyase.
Kirkwood
Mark
M
School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.
Le Brun
Nick E
NE
School of Chemistry, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.
Todd
Jonathan D
JD
School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.
Johnston
Andrew W B
AWB
School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK.
D000179:Acrylates / Q000378:metabolism; D013437:Carbon-Sulfur Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D020134:Catalytic Domain; D016297:Mutagenesis, Site-Directed; D041902:Rhodobacteraceae / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D013440:Sulfides / Q000378:metabolism; D013452:Sulfonium Compounds / Q000378:metabolism
D000179:Acrylates; D013440:Sulfides; D013452:Sulfonium Compounds; C068078:dimethylpropiothetin; D013437:Carbon-Sulfur Lyases; C400924:dimethylsulfoniopropionate lyase; C036658:acrylic acid; C004784:dimethyl sulfide