Abstract
[Objective] To purify and study the enzymatic properties of acid protease from the Trichoderma reesei strain, and to screen the stabilizer formula. [Methods] The acid protease is purified by ammonium sulfate precipitation, dialysis, and anion exchange column chromatography. Then, its molecular weight is determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and its enzymatic properties are studied. Finally, the stabilizer formula of acid protease is optimized, with an investigation into its effect on acid protease storage stability. [Results] For the purified acid protease, its specific enzyme activity is 4 849 U/mg, purification fold is 3.28 times, and relative molecular weight is 40.The optimal reaction condition of the enzyme is a pH of 3.0 and a temperature of 55 ℃, with satisfactory stability in the pH range of 3.0 to 6.0 and below 60 ℃. The enzyme activity retention rate is 69 % at 60 ℃, indicating high temperature resistance. In metal ions, Na+, Zn2+, and Mn2+ protect acid protease stability. Stored at a concentration of 20 % sodium chloride for 7 days, acid protease still retains certain activity, exhibiting high salt tolerance. The optimal formula of acid protease stabilizer is: maltose 26 %, mannitol 12.5 %, sodium chloride 12 %, and zinc chloride 0.1 mol/L. Added with stabilizer, the enzyme activity of acid protease is 92.8 % after storage at 45 ℃ for 48 days. [Conclusion] Acid protease is high in heat and salt resistance, while the stabilizer formula can significantly improve acid protease stability.
Publication Date
5-15-2026
First Page
26
Last Page
32
DOI
10.13652/j.spjx.1003.5788.2025.80561
Recommended Citation
Yao, WU; Huashun, YU; Rui, CHEN; Zhaoshi, YANG; Ming, LI; Jie, GAO; and Yan, ZHANG
(2026)
"Purification, enzymatic properties, and stabilizer screening of acid protease from Trichoderma reesei,"
Food and Machinery: Vol. 42:
Iss.
4, Article 4.
DOI: 10.13652/j.spjx.1003.5788.2025.80561
Available at:
https://www.ifoodmm.cn/journal/vol42/iss4/4
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