Purification, characterization and gene cloning of isoeugenol-degrading enzyme from Pseudomonas putida IE27
- 14 February 2007
- journal article
- research article
- Published by Springer Nature in Archiv für Mikrobiologie
- Vol. 187 (6) , 511-517
- https://doi.org/10.1007/s00203-007-0218-9
Abstract
An isoeugenol-degrading enzyme was purified to homogeneity from Pseudomonas putida IE27, an isoeugenol-assimilating bacterium. The purified enzyme was a 55 kDa monomer and catalyzed the initial step of isoeugenol degradation, the oxidative cleavage of the side chain double-bond of isoeugenol, to form vanillin. Another reaction product of isoeugenol degradation besides vanillin was identified to be acetaldehyde. The values of Km and k cat for isoeugenol were 175 μM and 5.18 s–1, respectively. The purified enzyme catalyzed the incorporation of an oxygen atom from either molecular oxygen or water into vanillin, suggesting that the isoeugenol-degrading enzyme is a kind of monooxygenase. The gene encoding the isoeugenol-degrading enzyme and its flanking regions were isolated from P. putida IE27. The amino acid sequence of the enzyme was similar to those of lignostilbene-α,β-dioxygenases, carotenoid monooxygenases and 9-cis-epoxycarotenoid dioxygenases.Keywords
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