猪血蛋白酪氨酸磷酸酶(PTP1B)的提取、纯化  

Isolation, Purification and Characterization of Porcine Blood Protein Tyrosine Phosphatase 1B

作　　者：闵玉涛 宋彦显 马庆一[2] 刘鑫[3] MIN Yutao;SONG Yanxian;MA Qingyi;LIU Xin(Zhengzhou Institute of Technology,Zhengzhou 450044;Zhengzhou University of Light Industry,Zhengzhou 450002;Hubei Zhongyan Industrial Co.,Ltd.Wuhan 430000)

机构地区：[1]郑州工程技术学院,郑州450044 [2]郑州轻工业学院,郑州450002 [3]湖北中烟工业有限责任公司,武汉430000

出　　处：《食品工业》2019年第9期80-84,共5页The Food Industry

基　　金：河南省2015年科技攻关计划项目(152102310010);河南省2017年高等学校青年骨干教师培养计划(2017GGJS184);河南省2017年高等学校重点科研项目计划(18A150050);郑州市2014年科技攻关计划项目(141PPTGG456)

摘　　要：以猪血为原料,采用丙酮沉淀、硫酸铵分级盐溶盐析、透析及反渗透等方法使之部分纯化,并对其性质进行研究。结果表明,硫酸铵分级盐溶盐析和透析相结合的方法纯化后的PTP1B酶活力明显提高,其中70%透析酶的比活力为434.5 U/mg,高于60%(402.1 U/mg)和80%(154.7 U/mg)时的比活力,是粗酶比活力的3.36倍,可作为试验用酶。纯化后的蛋白酪氨酸磷酸酶K_m为4.25 mmol/L。在35~42℃温度范围内保持较高的活力,其最适温度为37℃;在pH 5.50~7.67范围内保持较高的活力,其最适pH为7.20。反应速度与酶量、底物浓度的关系均符合酶促反应动力学。The Protein tyrosine phosphatase 1 B(PTP1 B) from the pig blood was extracted and purified by ammonium sulfate solution, salting out, dialysis and reverse osmosis, and its properties was investigated. The results showed that the enzyme specific activity were significantly increased by the extraction and purification conditions. The PTP1 B of 70% dialyse specific activity(434.5 U/mg) was as 3.36 times as that of the crude enzyme, higher than 60%(402.1 U/mg) and 80%(154.7 U/mg). The enzyme kinetics indicated K_m was 4.25 mmol/L. The PTP1 B was relatively stable in the range of 35–42 ℃ and pH 5.50–7.67. Its optimum temperature and pH were 37 ℃ and 7.20, respectively. The relationship between the reaction rate and the amount of enzyme and the concentration of substrate conformed to the enzymatic reaction kinetics.

关 键 词：蛋白酪氨酸磷酸酶 提取 纯化 

分 类 号：TS2[轻工技术与工程—食品科学与工程]