Purification and characterization of Aspergillus oryzae LK-101 salt-tolerant acid protease isolated from soybean paste

被引:21
作者
Lee, Si-Kyung [2 ]
Hwang, Joo-Yeon [2 ]
Choi, Seung Hwa [1 ]
Kim, Sang Moo [1 ]
机构
[1] Gangneung Wonju Natl Univ, Fac Marine Biosci & Technol, Kangnung 210702, Gangwon, South Korea
[2] Konkuk Univ, Dept Appl Biol & Chem, Seoul 143701, South Korea
关键词
Aspergillus oryzae; kinetics; salt-tolerant acid protease; soybean paste; stability; ALKALINE PROTEASE; SAUCE; ENZYME;
D O I
10.1007/s10068-010-0047-5
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
A novel salt-tolerant acid protease was produced from Aspergillus oryzae LK-101 (AOLK-101). The AOLK-101 protease was purified to homogeneity by ammonium sulfate precipitation, DEAE-Sephadex A-50 and Sephadex G-100 chromatographies in order. The specific activity and the purification ratio of the purified protease were 2,301 unit/mg and 11.6 fold, respectively, with 25 kDa of molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrpphoresis (SDS-PAGE). Its optimal pH and temperature were pH 6.5 and 50A degrees C, respectively. This protease was relatively stable at pH 4.5-7.5, below 40A degrees C, and up to 10% salt concentration. The protease was moderately inhibited by Ag2+ and Zn2+, and strongly by ethylenediamide tetraacetic acid (EDTA) and phenylmethysulfonyl fluoride (PMSF), but activated by Cu2+ and Mn2+. Therefore, the AOLK-101 protease was a serine protease based on the influence of metal ions and inhibitors. K (m) , V (max) , k (cat) , and k (cat) /K (m) values of AOLK-101 protease for hammastein milk casein were 1.04 mg/mL, 124.84 unit/L, 163.5/sec, and 3.9x10(6)/m center dot sec, respectively.
引用
收藏
页码:327 / 334
页数:8
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