trans-Dienelactone hydrolase from Pseudomonas reinekei MT1, a novel zinc-dependent hydrolase.
Pseudomonas reinekei MT1 is capable of growing on 4- and 5-chlorosalicylate, involving a pathway with trans-dienelactone hydrolase (trans-DLH) as a key enzyme. It acts on 4-chloromuconolactone formed during cycloisomerization of 3-chloromuconate by hydrolyzing it to maleylacetate. The gene encoding this activity was localized, sequenced and expressed in Escherichia coli. Inductively coupled plasma mass spectrometry showed that both the wild-type as well as recombinant enzymes contained 2 moles of zinc but variable amounts of manganese/mol of protein subunit. The inactive metal-free apoenzyme could be reactivated by Zn(2+) or Mn(2+). Thus, trans-DLH is a Zn(2+)-dependent hydrolase using halosubstituted muconolactones and trans-dienelactone as substrates, where Mn(2+) can substitute for Zn(2+). It is the first member of COG1878 and PF04199 for which a direct physiological function has been reported.
Cámara
Beatriz
B
Department of Microbial Pathogenesis, HZI-Helmholtz Centre for Infection Research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Marín
Macarena
M
Schlömann
Michael
M
Hecht
Hans-Jürgen
HJ
Junca
Howard
H
Pieper
Dietmar H
DH
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D002384:Catalysis; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008345:Manganese / Q000737:chemistry / Q000378:metabolism; D008667:Metalloproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D013379:Substrate Specificity; D015032:Zinc / Q000737:chemistry* / Q000378:metabolism
D001426:Bacterial Proteins; D008667:Metalloproteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D008345:Manganese; D002265:Carboxylic Ester Hydrolases; C046130:carboxymethylenebutenolidase; D015032:Zinc