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Biochemical studies on cloned Bacillus sp. BP-7 phenolic acid decarboxylase PadA

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Abstract

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°C and pH 5.5, being stable over a broad range of pH and up to 45°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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Acknowledgements

We thank the Serveis Cientifico-Tècnics of the University of Barcelona for technical aid in sequencing. This work was partially financed by the Scientific and Technological Research Council (CICYT, Spain), grants ALI97-0356-C02-02 and REN2001-3224, by the III Pla de Recerca de Catalunya (Generalitat de Catalunya), grant 2001SGR-00143, and by the Generalitat de Catalunya to the "Centre de Referència en Biotecnologia" (CeRBa).

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  1. Department of Microbiology, Faculty of Biology, University of Barcelona, Av. Diagonal 645, 08028, Barcelona, Spain

    N. Prim, F. I. J. Pastor & P. Diaz

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  1. N. Prim
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  2. F. I. J. Pastor
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  3. P. Diaz
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Correspondence to P. Diaz.

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Prim, N., Pastor, F.I.J. & Diaz, P. Biochemical studies on cloned Bacillus sp. BP-7 phenolic acid decarboxylase PadA. Appl Microbiol Biotechnol 63, 51–56 (2003). https://doi.org/10.1007/s00253-003-1371-y

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  • DOI: https://doi.org/10.1007/s00253-003-1371-y

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