Acid protease has wide applications in foods, brewing, feeds, and leather industries. However, current acid proteases are unstable when the temperature or pH values are higher than 50 ℃ or 3.0, respectively, which restricts their applications. Thus, acid protease encoding gene expA from Aspergillus niger CICIM F0510 was successfully cloned and expressed in Pichia pastoris, and the recombinant strain GS115 (pPIC-expA) was generated in this study. In shaking flask experiments, the activity of recombinant enzyme EXPA was 257 380 RFU/h. Bioinformatics analysis showed that this enzyme belonged to family A1A of aspartic proteases. The optimal temperature and pH of EXPA were 50 ℃ and 3.0, respectively. After incubating at 40-50 ℃ or pH 2.5-3.5 for 1 h, the recombinant enzyme still retained more than 80% of its maximum activity. Zn2+, Ca2+, Fe2+, and Mn2+ enhanced EXPA activity, whereas Cu2+, Co2+, Fe3+, EDTA, and SDS had significant inhibitory effects on EXPA activity. Besides, EXPA also hydrolyzed soybean protein isolates, water-soluble corn protein, and wheat protein. Good thermostability and pH stability of EXPA provide a solid foundation for its applications in food and feed industries.
WANG Xin
,
JIN Peng
,
Song Peng
,
Dong Zixing
,
LIU Xiaoguang
,
WANG Zhengxiang
. Cloning, expression and biochemical characterization of a novel acidprotease EXPA from Aspergillus niger[J]. Food and Fermentation Industries, 2019
, 45(3)
: 40
-46
.
DOI: 10.13995/j.cnki.11-1802/ts.018387
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