Biochemical studies on cloned Bacillus sp. BP-7 phenolic acid decarboxylase PadA.
Sequence analysis of a Bacillus sp. BP-7 recombinant clone coding for a previously described carboxylesterase revealed the presence of an additional ORF with homology to bacterial hydroxycinnamic acid decarboxylases. Analysis of the amino acid sequence of the encoded enzyme revealed the presence of a single, highly conserved domain of 161 amino acids, with a predicted molecular mass of 19,143 Da and a pI of 5.5. Crude cell extracts from the recombinant clone displayed activity on ferulic, p-coumaric and caffeic acids, with no need for added cofactors. The cloned enzyme, named PadA, displayed maximum activity at 40 degrees C and pH 5.5, being stable over a broad range of pH and up to 45 degrees C. HPLC analysis of the products of catalysis revealed the conversion of phenolic acids to their aromatic 4-vinyl derivatives, with no accumulation of other by-products. PadA was found as a homodimer in the parental Bacillus sp. BP-7 strain and its expression was induced by both hydroxycinnamic acids and their corresponding derivative products. The results obtained suggest that the enzyme could be involved in a stress response for conversion of toxic hydroxycinnamic acids released after plant cell wall degradation.
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Department of Microbiology, Faculty of Biology, University of Barcelona, Av. Diagonal 645, 08028 Barcelona, Spain.
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Diaz
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D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D002109:Caffeic Acids / Q000378:metabolism; D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003373:Coumaric Acids / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D011422:Propionates; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D002109:Caffeic Acids; D003373:Coumaric Acids; D004269:DNA, Bacterial; D011422:Propionates; C004999:ferulic acid; D002262:Carboxy-Lyases; C417714:phenolic acid decarboxylase; C495469:p-coumaric acid; C040048:caffeic acid