食品科学2017，Vol.38Issue(22)：34-40,7.DOI:10.7506/spkx1002-6630-201722006
凡纳滨对虾组织蛋白酶L性质分析及其对肌肉蛋白的降解
Characterization of Cathepsin L from Litopenaeus vannamei and Its Effect on Muscular Protein Degradation
摘要
Abstract
This study aimed to investigate the enzyme that might be involved in this process.Cathepsin L was purified to homogeneity from the hepatopancreas of Litopenaeus vannamei,and its molecular mass was approximately 31 kD as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).Peptide mass fingerprinting revealed that 8 peptide fragments with a total of 112 amino acid residues were completely identical to the cathepsin L gene of L.vannamei.Using Z-Phe-Arg-MCA as substrate,the proteinase revealed optimal activity at 35 ℃ and pH 5.5,respectively.Purified cathepsin L was stable at temperature up to 40 ℃ and in the pH range from 5.5 to 6.5.It exhibited high specificity towards the substrate Z-Phe-Arg-MCA.Its enzymatic activity was significantly suppressed by the cysteine proteinase inhibitors E-64 and leupeptin,while it could be slightly activated by the metalloproteinase inhibitors ethylene diamine tetraacetic acid (EDTA) and ethylene glycol tetraacetic acid (EGTA).In addition,the fibrous structure of shrimp meat was increasingly destroyed with the prolongation of cold storage time as observed by scanning electron microscope (SEM).Purified cathepsin L effectively hydrolyzed muscular proteins as detected by SDS-PAGE,suggesting its potential involvement in the degradation of shrimp muscular proteins during cold storage.关键词
凡纳滨对虾/组织蛋白酶L/纯化/肌原纤维/扫描电子显微镜/降解Key words
Litopenaeus vannamei/cathepsin L/purification/myofibillar protein/degradation/scanning electron microscope (SEM)分类
轻工纺织引用本文复制引用
颜龙杰,沈建东,张凌晶,翁凌,胡莉苹,曹敏杰..凡纳滨对虾组织蛋白酶L性质分析及其对肌肉蛋白的降解[J].食品科学,2017,38(22):34-40,7.基金项目
国家自然科学基金面上项目(31271838) （31271838）
厦门南方海洋研究中心项目(14CZP030HJ04) （14CZP030HJ04）
