食品科学 ›› 2005, Vol. 26 ›› Issue (7): 46-49.

• 基础研究 • 上一篇    下一篇

K.fragilis LFS-8611 β-D-半乳糖苷酶的分离纯化

 刘建福, 陈庆森, 王璋   

  1. 天津食品生物技术重点实验室,江南大学食品学院 天津商学院生物工程系
  • 出版日期:2005-07-15 发布日期:2011-09-19

Purification of K. fragilis LFS-8611 β-D-galactosidase

 LIU  Jian-Fu, CHEN  Qing-Sen, WANG  Zhang   

  1. 1.Tianjin Key Laboratory of Food Biotechnology, Tianjin University of Commerce;2.School of Food Science and Technology, Southern Yangtze University
  • Online:2005-07-15 Published:2011-09-19

摘要: 分别采用柱色谱技术对K.fragilisLFS-8611β-D-半乳糖苷酶进行了纯化,十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定了该酶的纯度。粗酶先后经过(NH4)2SO4分级沉淀、CM-SepharoseCL-6B、DEAE-SepharoseCL-6B、SephacrylS-200柱色谱四步分离纯化,酶的最终回收率为16%,纯化倍数为45.8。纯化的酶在SDS-PAGE上为一条染色谱带,相对分子质量约为60000。

关键词: &beta, -D-半乳糖苷酶, 纯化

Abstract: The β-D-galactosidase from K. fragilis LFS-8611 was purified by chromatography methods, and the purity ofenzyme was identified by sodium dodecyl sulphate polyacrylamide gel electrophoresis(SDS-PAGE). The enzyme was purified about 45.8 fold with a yield of 16% total activity by successive ammonium sulfate-fractionation, CM-Sepharose CL-6B, DEAE-Sepharose CL-6B, and Sephacryl S-200 column chromatography. When the purified enzyme samples were subjected to SDS-PAGE, one single band was observed, which suggested that the enzyme samples were almost pure. The molecular weightestimated for K. fragilis LFS-8611 β-D-galactosidase by PAGE was 60000.

Key words: D-galactosidase, purification