1 2 3 4 Uncovering the gallate decarboxylase involved in tannin degradation on 5 Lactobacillus plantarum WCFS1 6 7 8 Natalia Jiménez, José Antonio Curiel, Inés Reverón, Blanca de las Rivas, Rosario 9 Muñoz 10 11 12 Laboratorio de Biotecnología Bacteriana, Instituto de Ciencia y Tecnología de Alimentos y 13 Nutrición, ICTAN-CSIC, Juan de la Cierva 3, 28006 Madrid, Spain 14 15 16 17 Running Title 18 LACTOBACILLUS PLANTARUM GALLATE DECARBOXYLASE 19 20 Address correspondence to Rosario Muñoz,
[email protected] 21 1 22 Lactobacillus plantarum are lactic acid bacteria strains able to degrade tannins by the 23 subsequent action of tannase and gallate decarboxylase enzymes. The tannase 24 encoding gene had been previously identified, whereas the gene encoding gallate 25 decarboxylase was unknown. SDS-PAGE from gallic-acid induced L. plantarum 26 extracts showed a 54-kDa protein which was absent in the uninduced cells. This 27 protein was identified as Lp_2945, putatively annotated as UbiD. Homology searches 28 identified ubiD-like genes located within three-gene operons which encoded the three 29 subunits of non-oxidative aromatic acid decarboxylases. L. plantarum is the only 30 bacteria in which the lpdC gene (lp_2945) and the lpdB/lpdD genes are separated in the 31 chromosome (lp_0271/lp_0272). Combination of extracts from recombinant E. coli 32 cells expressing lpdB, lpdC, and lpdC genes, demonstrated that LpdC is the only 33 protein required to yield gallate decarboxylase activity. However, the disruption of 34 these genes in L. plantarum revealed that the lpdB and lpdC gene products are 35 essential for gallate decarboxylase activity.