PURIFICATION AND CHARACTERIZATION OF ACIDIC PROTEASE FROM ASPERGILLUS ORYZAE BCRC 30118

Authors

Li-Jung Yin, Department of Seafood, National Kaohsiung Marine University, Kaohsiung, Taiwan, R.O.C
Ya-Hui Chou, Department of Food Science, National Taiwan Ocean University, Keelung, Taiwan, R.O.C
Shann-Tzong Jiang, Department of Food Science, National Taiwan Ocean University, Keelung,Taiwan, R.O.C.Department of Food and Nutrition, Providence University, Taichung, Taiwan, R.O.C.Follow

Abstract

The acidic protease was purified from 4-day cultivation of Aspergillus oryzae BCRC 30118 by DEAE Sephacel and Sephacryl S-200 HR chromatographs. The specific activity, yield and purification fold were 117.62 kU/mg, 15.1% and 6.6, respectively. The molecular weight (M) was 41.0 kDa, while the optimal pH and temperature were 3.0 and 60°C, respectively. It was stable at pH 3.0-6.0 and 4-35°C. However, it was inhibited by Fe2+, Hg2+, Fe3+ and pepstatin A, and slightly by leupeptin and TPCK. According to substrate specificity and inhibitor study, it was a cysteine protease with activation energy of 37.5 kcal/mol. Its Km, Vmax, Kcat and Kcat/Km for the hydrolysis of hemoglobin were 0.12 mM, 14.29 µmol/min, 14.55 sec-1 and 125.80 (sec-1 mM-1), respectively

Recommended Citation

Yin, Li-Jung; Chou, Ya-Hui; and Jiang, Shann-Tzong (2013) "PURIFICATION AND CHARACTERIZATION OF ACIDIC PROTEASE FROM ASPERGILLUS ORYZAE BCRC 30118," Journal of Marine Science and Technology: Vol. 21: Iss. 1, Article 14.
DOI: 10.6119/JMST-012-0529-1
Available at: https://jmstt.ntou.edu.tw/journal/vol21/iss1/14