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Applied and Environmental Microbiology, August 2000, p. 3368-3375, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Knockout of the p-Coumarate Decarboxylase Gene from Lactobacillus plantarum Reveals the Existence of Two Other Inducible Enzymatic Activities Involved in Phenolic Acid Metabolism

Lise Barthelmebs, Charles Divies, and Jean-François Cavin*

Laboratoire de Microbiologie UMR-INRA, ENSBANA, Université de Bourgogne, 21000 Dijon, France

Received 22 February 2000/Accepted 27 May 2000

Lactobacillus plantarum NC8 contains a pdc gene coding for p-coumaric acid decarboxylase activity (PDC). A food grade mutant, designated LPD1, in which the chromosomal pdc gene was replaced with the deleted pdc gene copy, was obtained by a two-step homologous recombination process using an unstable replicative vector. The LPD1 mutant strain remained able to weakly metabolize p-coumaric and ferulic acids into vinyl derivatives or into substituted phenyl propionic acids. We have shown that L. plantarum has a second acid phenol decarboxylase enzyme, better induced with ferulic acid than with p-coumaric acid, which also displays inducible acid phenol reductase activity that is mostly active when glucose is added. Those two enzymatic activities are in competition for p-coumaric and ferulic acid degradation, and the ratio of the corresponding derivatives depends on induction conditions. Moreover, PDC appeared to decarboxylate ferulic acid in vitro with a specific activity of about 10 nmol · min-1 · mg-1 in the presence of ammonium sulfate. Finally, PDC activity was shown to confer a selective advantage on LPNC8 grown in acidic media supplemented with p-coumaric acid, compared to the LPD1 mutant devoid of PDC activity.

* Corresponding author. Mailing address: Laboratoire de Microbiologie UMR-INRA, ENSBANA, Université de Bourgogne, 1 esplanade Erasme, F-21000 Dijon, France. Phone: (33) 03 80 39 66 72. Fax: (33) 03 80 39 66 40. E-mail: cavinjf{at}u-bourgogne.fr.

Applied and Environmental Microbiology, August 2000, p. 3368-3375, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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