食品科学

• 生物工程 • 上一篇    下一篇

海参体壁中α-1,4淀粉酶的分离纯化及性质

张 杰,张永勤*,罗彩华,刘征东,程袁芬,卢菲菲,张 童   

  1. 青岛科技大学化工学院,山东 青岛 266042
  • 出版日期:2015-03-15 发布日期:2015-03-17

Purification and Characterization of α-1,4-Amylase in Body Wall of Sea Cucumber

ZHANG Jie, ZHANG Yongqin*, LUO Caihua, LIU Zhengdong, CHENG Yuanfen, LU Feifei, ZHANG Tong   

  1. College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266042, China
  • Online:2015-03-15 Published:2015-03-17

摘要:

通过采用匀浆、硫酸铵盐析、透析、DEAE-52阴离子交换层析、Sephacryl S-300凝胶层析等方法从海参体壁中分离纯化并得到了α-1,4淀粉酶。结果表明,该酶具有3 条亚基链,分子质量约为420 kD;最适pH值为9.0,最适温度为80 ℃,90 ℃条件下保温30 min后仍有38%以上的相对酶活性,因此,该酶是一种具有较高热稳定性的碱性高温淀粉酶。K+、Fe3+和Mn2+对其有较强的抑制作用,Cu2+和Ca2+对其有较强的激活作用。

关键词: 海参, 自溶, 淀粉酶, 分离, 纯化, 热稳定性

Abstract:

α-1,4-Amylase was extracted and isolated from the body wall of sea cucumber by homogenization, salting out
with ammonium sulfate, dialysis, DEAE-52 anion exchange chromatography and Sephacryl S-300 gel chromatography
sequentially. The results showed that the purified enzyme had three subunit chains according to the SDS-PAGE, with
molecular weight of 420 kD; the optimum pH and temperature were 9.0 and 80 ℃; and the residual activity remained at least
38% after inactivation at 90 ℃ for 30 min. Therefore, this enzyme is a thermostable alkaline amylase. This amylase could be
strongly activated by K+, Fe3+ or Mn2+ and inhibited by Cu2+ or Ca2+.

Key words: sea cucumber, autolysis, amylase, isolation, purification, thermostability

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