食品科学 ›› 2007, Vol. 28 ›› Issue (12): 278-282.

• 生物工程 • 上一篇    下一篇

重组醇醛脱氢酶的分离纯化及性质研究

 谢莉, 张铎, 郭会灿, 张丽萍, 李曼, 赵宝华   

  1. 河北师范大学生命科学院; 石家庄职业技术学院; 河北省生物研究所; 河北师范大学生命科学院 河北石家庄050016; 河北石家庄050016; 河北石家庄050081;
  • 出版日期:2007-12-15 发布日期:2011-11-22

Purification and Specific Properties of Recombinant L-sorbose/L-sorbosone Dehydrogenase

 XIE  Li, ZHANG  Duo, GUO  Hui-Can, ZHANG  Li-Ping, LI  Man, ZHAO  Bao-Hua   

  1. 1.College of Life Science,Hebei Normal University, Shijiazhuang 050016, China; 2.Shijiazhuang Vocational Technology Institute, Shijiazhuang 050081, China; 3.Hebei Institute of Biology, Shijiazhuang 050081, China
  • Online:2007-12-15 Published:2011-11-22

摘要: 基因工程菌Y517#能表达醇醛脱氢酶,将L-山梨糖转化为VC的前体2-酮基-古龙酸(2-KGA)。通过超声波破碎菌体、硫酸铵分级沉淀、DEAE Sepharose FastFlow阴离子交换层析,Q Sepharose High Performance柱层析等过程,从Y517#发酵液中分离纯化了重组醇醛脱氢酶,纯化倍数为11倍。研究发现,该酶分子量为66kD,最适作用温度为40℃,对热不稳定,在60℃下保温10min酶活力完全丧失。最适pH值为7.0,pH稳定范围为6.0~9.0。

关键词: 重组醇醛脱氢酶, 基因工程菌, 分离纯化, 酶学性质

Abstract: L-sorbose/L-sorbosone dehydrogenase produced by gene engineering strain Y517# can transform L-sorbose to 2-KGA, the Vitamin C precursor. By means of ammonium sulfate precipitation, DEAE Sepharose Fast Flow and Q Sepharose High Performance assay, a purified recombinant L-sorbose/L-sorbosone dehydrogenase was obtained from Y517#. The results showed that the molecular weight of the purified enzyme is of 66 kD. The optimum temperature and pH are 40 ℃ and 7.0 respectivily. The enzyme activity is stable at pH 6.0~9.0 and below 45℃.

Key words: recombinant L-sorbose/L-sorbosone dehydrogenase, gene engineering strain, purification, enzyme properties