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

Protein Expr Purif. 2019 May:157:36-41. doi: 10.1016/j.pep.2019.01.005. Epub 2019 Jan 9.

Abstract

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.

Keywords: Gallic acid; Lactic acid bacteria; Lactobacillus pentosus; Miang; Tannase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carboxylic Ester Hydrolases / genetics
  • Carboxylic Ester Hydrolases / metabolism*
  • Cloning, Molecular / methods*
  • Escherichia coli / genetics
  • Gallic Acid / metabolism
  • Hydrogen-Ion Concentration
  • Lactobacillus pentosus / enzymology*
  • Lactobacillus pentosus / genetics
  • Lactobacillus pentosus / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Tannins / metabolism
  • Temperature

Substances

  • Recombinant Proteins
  • Tannins
  • Gallic Acid
  • Carboxylic Ester Hydrolases
  • tannase