Identification and cloning of a gene encoding tannase (tannin acylhydrolase) from Lactobacillus plantarum ATCC 14917T

Abstract

The gene tanLpl, encoding a novel tannase enzyme (TanLpl), has been cloned from Lactobacillus plantarum ATCC 14917(T). This is the first report of a tannase gene cloned from a bacterial source other than from Staphylococcus lugdunensis, which has been reported elsewhere. The open reading frame of tanLpl, spanning 1410 bp, encoded a 469-amino-acid protein that showed 28.8% identity to the tannase of S. lugdunensis with several commonly conserved sequences. These sequences could not be found in putative tannases reported for other bacteria and fungi. TanLpl was expressed in Escherichia coli DH5alpha from a pGEM-T expression system and purified. SDS-PAGE analysis indicated that purified TanLpl was a monomer polypeptide of approximately 50 kDa in size. Subsequent enzymatic characterization revealed that TanLpl was most active in an alkaline pH range at 40 degrees C, which was quite different from that observed for a fungal tannase of Aspergillus oryzae. In addition, the Michaelis-Menten constant of TanLpl was markedly lower than that of A. oryzae tannase. The evidence suggests that TanLpl should be classified into a novel family of tannases.

Tannin Degradation by a Novel Tannase Enzyme Present in Some Lactobacillus plantarum Strains

“…In addition, the optimal temperature and pH for TanA Lp (20 to 30°C and pH 6) differed from those described for TanB Lp (40°C and pH 7 to 8) (7,8). Interestingly, the optimal activity of TanA Lp is similar to the optimal tannase activity shown by cell extracts from L. plantarum ATCC 14917…”

“…Despite the extensive interest and long history of the study of tannase, there is surprisingly little knowledge about the enzyme at the molecular level, which has become one of the critical factors that limit the large-scale application of tannase. To our knowledge, the only bacterial tannases that have been analyzed genetically are those from Staphylococcus lugdunensis (6), Lactobacillus plantarum (7,8), and Enterobacter sp. (9).…”

“…(9). In addition, L. plantarum tannase has been biochemically and structurally characterized (7,8,10).…”

“…The biochemical pathway for the degradation of tannins by L. plantarum involves the action of a tannase and a gallate decarboxylase to decarboxylate the gallic acid formed by tannase action (14-16). The L. plantarum genes encoding tannase (tanB Lp , formerly called tanLp1) (7) and gallate decarboxylase (lpdBCD) (16) involved in tannin degradation have been identified. However, an additional putative L. plantarum tannase sequence has been annotated in the genome of an L. plantarum strain.…”

Digestion of Tannin by Bacteria Enterobacter cloacae from the Gut of Indian Mole Cricket (Gryllotalpa krishnani)

“…Previously, Lactobacillus plantarum and K. pneumonia tannase were also found to be a monomer with a molecular mass of 46.5 and 50 kDa, respectively [37,38]. In insects the digestion of food by gut associated bacteria has been demonstrated in cricket's mole, germ free crickets have much less hydrolytic enzymes activity a conventional cousin and these bacteria help in utilizing a wide range of food substrate [22].…”