食品科学 ›› 2012, Vol. 33 ›› Issue (3): 168-173.doi: 10.7506/spkx1002-6630-201203035

• 生物工程 • 上一篇    下一篇

猪背最长肌焦磷酸酶的分离纯化与酶学特性研究

靳红果,万可慧,田锐花,彭增起,王蓉蓉   

  1. 南京农业大学 教育部肉品加工与质量控制重点实验室
  • 收稿日期:2017-02-24 修回日期:2017-02-24 出版日期:2012-02-15 发布日期:2012-02-14

Purification and Characterization of Pyrophosphatase from Pork longissimus dorsi Muscle

JIN Hong-guo,WAN Ke-hui,TIAN Rui-hua,PENG Zeng-qi,WANG Rong-rong   

  1. Key Laboratory of Animal Products Processing and Quality Control, Ministry of Education, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2017-02-24 Revised:2017-02-24 Online:2012-02-15 Published:2012-02-14

摘要: 通过离心、50%~70%饱和硫酸铵沉淀、DEAE-52离子交换柱层析,从猪背最长肌中分离纯化出焦磷酸酶(PPase)。变性聚丙烯酰胺凝胶电泳图谱显示,PPase分子质量约72kD。焦磷酸酶酶学特性研究表明,最适反应温度和pH值分别为50℃和7.5。Mg2+是PPase的激活剂,在浓度4.75mmol/L时,酶活力最强。但Na+和K+都能抑制酶的活力,且Na+的抑制效果强于K+。PPase水解焦磷酸钠(TSPP)的动力学参数Vmax为0.086μmol /(L·min)),Km为0.36mmol/L。

关键词: 焦磷酸酶, 纯化, 酶学特性, 猪肉

Abstract: Pyrophosphatase (PPase) from pork longissimus dorsi muscle was purified by ultracentrifugation, 50%-70% saturated ammonium sulfate fractionation, DEAE-52 anion-exchange chromatography. The purified enzyme with molecular mass of 72 kD ran as a single band on SDS-polyacrylamide gel. The optimum pH and temperature for the isolated PPase was 7.5 and 50 ℃, respectively. Mg2+ was necessary for PPase and the activity reached maximum at a concentration of 4.75 mmol/L. Na+ and K+ inhibited the enzyme activity and the inhibitory effect of Na+ was stronger than K+. The kinetic constant Km and Vmax using TSPP as substrate were determined as 0.36 mmol/L and 0.086μmol/(L·min), respectively.

Key words: pyrophosphatase, purification, characterization, pork

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