蜡状芽孢杆菌ZJB-07112酰胺酶的分离纯化及其酶学性质  

Purification and characterization of amidase from Bacillus cereus ZJB-07112

作　　者：张俊伟[1] 郑裕国[1] 沈寅初[1] 

机构地区：[1]浙江工业大学生物工程研究所,浙江杭州310014

出　　处：《化工学报》2008年第3期624-629,共6页CIESC Journal

基　　金：国家重点基础研究发展计划项目(2003CB716005)~~

摘　　要：An amidase from a strain Bacillus cereus ZJB-07112 was purified to homogeneity by using sonication, anion-exchange chromatography, phenyl-sepharose chromatography.The molecular weight of amidase was estimated to be 60.6×103 by 12.5% SDS-PAGE.Its N-terminal sequence was ATIRPDDKAI.The optimum pH and temperature of the amidase for acrylamide were pH 7.5 and 35℃, respectively.The enzyme was unstable at a temperature over 50℃ and only 10.8% activity was retained after exposure to 60℃ for 30 min.Most of metal ions and EDTA had no significant effect on the enzyme activity, whereas Hg+,Ag+ and urea caused obvious inhibition.The Km and Vmax values of the amidase for acrylamide were 2.64 mmol·L-1 and 0.6 μmol·min-1·ml-1, respectively.An amidase from a strain Bacillus cereus ZJB-07112 was purified to homogeneity by using sonication, anion-exchange chromatography, phenyl-sepharose chromatography. The molecular weight of amidase was estimated to be 60.6×10^3 by 12.5% SDS-PAGE. Its N-terminal sequence was ATIRPDDKAI. The optimum pH and temperature of the amidase for acrylamide were pH 7.5 and 35℃, respectively. The enzyme was unstable at a temperature over 50℃ and only 10.8% activity was retained after exposure to 60℃ for 30 min. Most of metal ions and EDTA had no significant effect on the enzyme activity, whereas Hg^＋ , Ag^＋ and urea caused obvious inhibition. The Km and Vmax values of the amidase for acrylamide were 2.64 mmol·L^-1 and 0.6μmol·min^-1· ml^-1 , respectively.

关 键 词：蜡状芽孢杆菌 酰胺酶 丙烯酰胺 纯化 

分 类 号：Q814[生物学—生物工程]