食品科学

• 生物工程 • 上一篇    下一篇

红佳酿酵母β-葡萄糖苷酶的分离纯化及酶学性质

祝 霞,盛文军,杨学山,王 婧,李 敏,张 波,翦 祎,韩舜愈   

  1. 1.甘肃农业大学食品科学与工程学院,甘肃 兰州 730070;2.甘肃农业大学生命科学技术学院,甘肃 兰州 730070
  • 出版日期:2014-07-15 发布日期:2014-07-18

Purification and Enzymatic Characterization of β-Glucosidasein from Vintage Red Yeast

ZHU Xia, SHENG Wen-jun, YANG Xue-shan, WANG Jing, LI Min, ZHANG Bo, JIAN Yi, HAN Shun-yu   

  1. 1. College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China;
    2. College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
  • Online:2014-07-15 Published:2014-07-18

摘要:

以红佳酿酿酒酵母为出发菌株,通过离子交换法分离纯化β-葡萄糖苷酶,测定酶活力和蛋白质含量,并研究温度、pH值稳定性和金属离子、葡萄糖、酒精度对酶活力的影响。结果表明:粗酶液经过离子交换层析后,纯化倍数为19.41,得率为38.67%;经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamidegelelectrophoresis,SDS-PAGE)电泳检测为1 条谱带,达到电泳纯,分子质量约为45 kD;β-葡萄糖苷酶热稳定性较差,在20~40 ℃较稳定,在pH 5.0~10.0较稳定;Al3+、Cu2+ 对酶活力有抑制作用,K+、Mg2+、Ca2+、Zn2+、Na+对酶活力的影响不明显;葡萄糖和酒精对酶活力无明显抑制作用。

关键词: 红佳酿酵母, &beta, -葡萄糖苷酶, 分离纯化, 酶学性质

Abstract:

An intracellular β-glucosidase from Vintage Red Yeast was purified by ion exchange column chromatography.
The effects of temperature, pH, metal ions, glucose and alcohol on the stability of the purified β-glucosidase were studied.
The results showed that the enzyme was purified to 19.41 folds with a yield of 38.67%. The purity of β-glucosidase was
determined by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE). One band was obtained with a
molecular mass of about 45 kD. Characterization of the purified β-glucosidase indicated good thermostability in the range
from 20 to 40℃ and good pH stability in the range from 5.0 to 10.0. The enzyme activity was strongly inhibited by Al3+, and
Cu2+, whereas K+, Mg2+, Ca2+, Zn2+ and Na+ had no effect on the β-glucosidase activity. Glucose and alcohol did not have
significant inhibitory effect on the β-glucosidase activity.

Key words: vintage red yeast, β-glucosidase, purification, enzymatic characterization