| 南美白对虾丝氨酸蛋白酶的分离纯化及性质研究 |
| |
| 引用本文： | 翁凌,李腾,阴利华,孙乐常,苏文金,曹敏杰.南美白对虾丝氨酸蛋白酶的分离纯化及性质研究[J].厦门水产学院学报,2010(4):272-278. |
| |
| 作者姓名： | 翁凌 李腾 阴利华 孙乐常 苏文金 曹敏杰 |
| |
| 作者单位： | [1]集美大学生物工程学院,福建厦门361021 [2]上海海洋大学食品学院,上海201306 |
| |
| 基金项目： | 十一五国家科技支撑计划重大项目（2008BAD94B01）;福建省高校水产科学技术与食品安全重点实验室基金项目（2008J401） |
| |
| 摘 要： | 通过硫酸铵盐析、DEAE-Sepharose、Phenyl-Sepharose、Hydroxyapatite、Superdex75等方法,从南美白对虾消化腺中分离得到一种丝氨酸蛋白酶.SDS-PAGE结果显示,其分子质量约为28ku,最适pH值与最适温度分别为9.0和40℃,且pH值在7.0～10.0之间以及温度在40℃以下有较高的稳定性.底物特异性实验与抑制剂实验结果表明,该酶属于类胰蛋白酶的丝氨酸蛋白酶.动力学实验显示,以Boc-Phe-Ser-Arg-MCA为底物时,Km=0.69μmol/L,Kcat=0.33S-1,Kcat/Km=4.78×105（mol/L）-1S-1.
|
| 关 键 词： | 丝氨酸蛋白酶 南美白对虾 纯化 性质分析 |
Purification and Characterization of Serine Proteinase from Pacific White Shrimp |
| |
| Authors: | WENG Ling LI Teng YIN Li-hua SUN Le-chang SU Wen-jin CAO Min-jie |
| |
| Institution: | 1 (1. School of Biotechnology Engineering,Jimei University,Xiamen 361021,China; 2. College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China) |
| |
| Abstract: | A serine proteinase from the hepatopancreas of Pacific white shrimp was purified by a series of procedures,including ammonium sulfate precipitation,column chromatographies on DEAE-Sepharose, Phenyl-Sepharose,Hydroxyapatite and Superdex 75. Purified serine proteinase revealed a single band on SDS-PAGE. The molecular weight was about 28 ku,The optimum pH and temperature of the enzyme were 9. 0 and 40 ℃,respectivety. The enzyme was stable up to 35 ℃ and in the pH range from 7. 0 to 10. 0. Substrate specificity and inhibitor sensitivity experiments suggested that the enzyme was a trypsin-type serine proteinase. Kinetic constants of Km was 0. 69 μmol/L,Kcat was 0. 33 S-1 and Kcat /Km was 4. 78 × 105 （mol/ L）-1S-1 using Boc-Phe-Ser-Arg-MCA as substrate. |
| |
| Keywords: | serine proteinase Pacific white shrimp purification characterization analysis |
| 本文献已被 维普 等数据库收录！ |
|
