食品科学

• 生物工程 • 上一篇    下一篇

牛肾溶菌酶的分离纯化及部分酶学性质

傅 婷,万 骥,王 丹,唐云明*   

  1. 西南大学生命科学学院,淡水鱼类资源与生殖发育教育部重点实验室,三峡库区生态环境教育部重点实验室,重庆 400715
  • 出版日期:2016-03-15 发布日期:2016-03-17

Isolation, Purification and Partial Characterization of Lysozyme from Bovine Kidney

FU Ting, WAN Ji, WANG Dan, TANG Yunming*   

  1. Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, School of Life Science, Southwest University, Chongqing 400715, China
  • Online:2016-03-15 Published:2016-03-17

摘要:

将新鲜牛肾匀浆后,经由缓冲液提取,正丁醇除脂,硫酸铵分级沉淀,CM-Sepharose阳离子交换层析,Superdex-200凝胶过滤层析后得到电泳纯的牛肾溶菌酶。结果表明:该酶比活力为15 145.63 U/mg,回收率为29.13%,纯化倍数为231.05;分子质量约12.66 kD,为单亚基酶。最适反应温度为65 ℃,在65 ℃以下较稳定;最适pH 9.0,pH值在3.0~9.0范围内稳定性较好;测得最适条件下牛肾溶菌酶对溶壁微球菌的Km值为0.399 μg/mL;甲醇、乙醇、异丙醇和硫氰化钾(KSCN)对该酶有激活作用;十二烷基硫酸钠、Pb2+、Ag+对该酶有明显抑制作用。

关键词: 牛肾, 溶菌酶, 分离纯化, 酶学性质

Abstract:

Electrophoretically pure lysozyme from bovine kidney was obtained through homogenization, buffer extraction,
butanol degreasing, ammonium sulfate fractionation precipitation, CM-Sepharose ion-exchange chromatography and
Superdex-200 gel filtration chromatography. Results showed that the specific activity of the purified lysozyme was
15 145.63 U/mg with a recovery rate of 29.13% and the enzyme was purified 231.05 folds. The molecular weight of the
lysozyme was approximately 12.66 kD, which consisted of a single subunit. The optimum temperature of the enzyme was
65 ℃ and it was stable at temperatures below 65 ℃; the optimum pH of this enzyme was 9.0 and it was stable in the range
of pH 3.0–9.0. Its Km towards micrococcus lysodeikticus was 0.399 μg/mL. The enzyme activity was enhanced by methanol,
ethanol, isopropanol and KSCN, and inhibited by SDS, Pb2+, and Ag+.

Key words: bovine kidney, lysozyme, isolation and purification, enzymatic properties

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