樱桃谷鸭胸肉脂肪氧合酶的分离纯化及其酶学特性研究

doi:10.7506/spkx1002-6630-201307035

食品科学 ›› 2013, Vol. 34 ›› Issue (7): 166-170.doi: 10.7506/spkx1002-6630-201307035

樱桃谷鸭胸肉脂肪氧合酶的分离纯化及其酶学特性研究

何立超1，赵见营2，田甜2，章建浩2,*

1.华中农业大学楚天学院，食品与生物科技学院，湖北武汉 430205； 2.国家肉品质量安全控制工程技术研究中心；教育部肉品加工与质量控制重点实验室；农业部农畜产品加工与质量控制重点开放实验室；南京农业大学食品科技学院，江苏南京 210095

收稿日期:2012-03-04 修回日期:2013-02-28 出版日期:2013-04-15 发布日期:2013-03-20
通讯作者: 章建浩 E-mail:nau_zjh@njau.edu.cn
基金资助:
“十二五”国家科技支撑计划项目(2012BAD28B01)；国家公益性行业(农业)科研专项经费项目(201303082-2)；江苏高校优势学科建设工程资助项目

Purification and Partial Characterization of Lipoxygenase in Cherry-valley Duck Breast Muscle

HE Li-chao1，ZHAO Jian-ying2，TIAN Tian2，ZHANG Jian-hao2,*

1. College of Food and Biotechnology, Chutian College, Huazhong Agricultural University, Wuhan 430205, China； 2. National Center of Meat Quality and Safety Control, Key Laboratory of Meat Processing and Quality Control, Ministry of Education, Key Laboratory of Animal Products Processing and Quality Control, Ministry of Agriculture, College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Received:2012-03-04 Revised:2013-02-28 Online:2013-04-15 Published:2013-03-20
Contact: ZHANG Jian-hao E-mail:nau_zjh@njau.edu.cn

摘要/Abstract

摘要： 用硫酸铵分级沉淀和DEAE-Sepharose柱层析法分离纯化樱桃谷鸭胸肉中脂肪氧合酶(lipoxygenase，LOX)，同时对酶学性质进行研究。结果显示：在DEAE-Sepharose柱层析时，当NaCl洗脱浓度在0.3～0.35mmol/L时，LOX被分离出，LOX经过分离纯化后其纯化倍数可达到54.3，SDS-PAGE电泳显示其分子质量约为96kD；粗酶和纯酶性质略有不同；以亚油酸为底物，粗酶和纯酶的最适底物浓度分别为6.4、10mmol/L，最适反应温度分别为40、30℃，最适pH值分别是5.0、5.5；纯酶Km=1.139mmol/L、Vmax=0.07608U/min，纯酶与底物的亲和能力大于粗酶。

关键词: 樱桃谷鸭, 脂肪氧合酶, 分离纯化, 酶学特性

Abstract: Crude lipoxygenase (LOX) from Cherry-valley duck breast muscle was purified through 20%－40% ammonium sulfate fractionation and DEAE-Sepharose column chromatography. LOX was separated by gradient elution of DEAESepharose column with 0.3－0.35 mmol/L NaCl solution, resulting in a purification factor of 54.3. The molecular weight of the purified LOX was measured by SDS-PAGE to be 96 kD. Slight differences in enzymatic properties between the crude LOX and the purified LOX were observed. Using linoleic acid as the substrate, the optimal substrate concentrations, reaction temperatures and pH values were 6.4 mmol/L and 10 mmol/L, 40 ℃ and 30 ℃, and 5.0 and 5.5, respectively, and the Michaelis constant (Km) and maximum velocity (Vmax) for the purified LOX were 1.139 mmol/L and 0.07608 U/min, respectively. The purified LOX had a higher affinity for the substrate than the crude LOX.

Key words: Cherry-valley duck, lipoxygenase (LOX), isolation and purification, enzymatic characteristics

中图分类号:

Q814.1

何立超1，赵见营2，田甜2，章建浩2,*. 樱桃谷鸭胸肉脂肪氧合酶的分离纯化及其酶学特性研究[J]. 食品科学, 2013, 34(7): 166-170.

HE Li-chao1，ZHAO Jian-ying2，TIAN Tian2，ZHANG Jian-hao2,*. Purification and Partial Characterization of Lipoxygenase in Cherry-valley Duck Breast Muscle[J]. FOOD SCIENCE, 2013, 34(7): 166-170.

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樱桃谷鸭胸肉脂肪氧合酶的分离纯化及其酶学特性研究

Purification and Partial Characterization of Lipoxygenase in Cherry-valley Duck Breast Muscle

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