The practical use of tannase in tea beverage and pharmaceutical industries has been limited by a number of factors, including insufficient knowledge about its properties and high production costs. In the present study, gene tahA encoding tannase of Aspergillus niger CICIM F0510 was successfully cloned and expressed in Pichia pastoris, generating the recombinant strain GS115 (pPIC-tahA). In shaking flask experiments, the activity of the recombinant enzyme was 1.04 U/mL. The temperature and pH optima of TahA was shown to be 30 ℃ and 4.5, respectively. After incubation at 20-50 ℃ or pH 3.5-5.0 for 1 h and 2 h, respectively, the recombinant enzyme still retained more than 60% of its initial activity. Cu2+, Mn2+ and K+ significantly enhanced the activity of TahA, whereas Co2+, Fe3+, Ca2+, Mg2+, Zn2+, Sn2+, EDTA and SDS exerted different inhibitory effects on its activity. The Vmax and Km of TahA toward methyl gallate were determined to be 0.436 μmol/(mL·min) and 5.143 mmol/L. Besides, TahA hydrolyzed the ester catechins in tea extracts. All these distinct biochemical properties lay a solid foundation for the application of TahA in tea beverages.
LI Meng-di
,
ZHANG Zhi-meng
,
DONG Zi-xing
,
TIAN Kang-ming
,
JIN Peng
,
LIU Xiao-guang
,
WANG Zheng-xiang
. Cloning, expression and biochemical characterization of tannase TahA from Aspergillus niger[J]. Food and Fermentation Industries, 2018
, 44(11)
: 15
-21
.
DOI: 10.13995/j.cnki.11-1802/ts.017644
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