Expression and biochemical characterization of a new alkaline tannase from Lactobacillus pentosus

Lactobacillus pentosus BA-7 and L. pentosus QA1-5 are tannin-tolerant lactic acid bacteria that were isolated from Miang, a traditional fermented tea-leaf found in northern Thailand and a tannin-rich substrate. Tannase encoding genes were isolated, cloned and overexpressed in Escherichia coli BL21(DE3). The recombinant tannase was produced with production yields of 40 and 39 KU/L for LpTanBA-7 and LpTanQA1-5, respectively. Both revealed the same molecular weight of 50 kDa as estimated by SDS-PAGE and were optimally active under alkaline pH conditions LpTanQA1-5 revealed optimal temperatures in a range of 37-40 °C as is typically found in lactic acid bacteria, while LpTanBA-7 was active at higher temperatures with an optimum temperature range of 45-55 °C. LpTanBA-7 was found to be more stable within the same range of temperatures than LpTanQA1-5. Furthermore, it was active and stable toward various organic solvents and produced 50 mg/mL of gallic acid from 100 mg/mL tannic acid. Based on the results, LpTanBA-7 is considered a new alkali-moderately thermophilic tannase obtained from lactic acid bacterium that may be capable of a feasible production capacity of gallic acid and its esters. Furthermore, tannase that is active at high temperatures could also be used in tea products in order to develop a sweet aftertaste, as well as to improve levels of antioxidant activity.