中国食品学报2012，Vol.12Issue(6)：22-29,8.
牙鲆肌肉赖氨酸氨肽酶的分离纯化与性质研究
Purification and Characterization of an Aminopeptidase with Highest Preference for Lysine from Paralichthys Olivaceus Skeletal Muscle
摘要
Abstract
An aminopeptidase was purified from Paralichthys olivaceus skeletal muscle to homogeneity by ammonium sulfate fractionation and three column chromatographies, including DEAE -Sephacel, Phenyl -Sepharose, and DEAE -Sepharose Fast Flow. The molecular mass of the enzyme was approximately 100 ku. Optimum temperature and pH of the enzyme were 45 ℃ and 7.5, respectively. Metal-chelating agents such as EDTA, EGTA and 1,10-phenanthroline effectively inhibited the enzyme activity. Zn2+, Mn2+ and (or) Co2+ were quite possibly its metal cofactor(s), strongly suggesting the enzyme belong to metalloproteinase family. The enzyme purified in the present study was regarded as a lysine aminopeptidase (LysAP) according to its substrate specificity. Western blot analysis revealed that leucine aminopeptidase from red sea bream and LysAP from Paralichthys olivaceus have high homogeneity in amino acid sequences. Furthermore, LysAP distributes in tissues including skeletal muscle, heart, liver, spleen, stomach, kidney, gut and gill.关键词
牙鲆/赖氨酸氨肽酶/纯化/金属离子/免疫印迹Key words
Paralichthys olivaceus/ lysine aminopeptidase/ purification/ metal ions/ Western blot引用本文复制引用
陈曦,蔡秋凤,刘光明,苏文金,曹敏杰..牙鲆肌肉赖氨酸氨肽酶的分离纯化与性质研究[J].中国食品学报,2012,12(6):22-29,8.基金项目
国家自然科学基金项目(20872049,30871947) （20872049,30871947）
