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| 黄瓜过氧化物酶的分离纯化及酶学性质 | |
| 引用本文： | 胡瑞斌,李星,王红杨,王洁,唐云明. 黄瓜过氧化物酶的分离纯化及酶学性质[J]. 食品科学, 2014, 35(11): 168-173. DOI: 10.7506/spkx1002-6630-201411034 |
| 作者姓名： | 胡瑞斌 李星 王红杨 王洁 唐云明 |
| 作者单位： | 西南大学生命科学学院，重庆市甘薯工程研究中心，三峡库区生态环境教育部重点实验室，重庆 400715 |
| 基金项目： | 重庆市科委重点攻关项目(CSTC2011AB1027) |
| 摘 要： | 新鲜的黄瓜经匀浆、抽提、硫酸铵沉淀、CM-Sepharose 离子交换层析、Superdex-200凝胶过滤层析后获得电泳纯的过氧化物酶。该酶的比活力、回收率及纯化倍数分别为64 177.67 U/mg、9.58%、61.93。该酶的分子质量为41.15 kD，亚基分子质量为40.21 kD。该酶的最适温度和最适pH值分别为60 ℃和6。在25～45 ℃及pH 5～9的范围内非常的稳定。在测定条件下测得该酶的Km值为53.79 mmol/L。硫氰化钾对该酶活力基本无影响。尿素、K+、Mn2+、Ca2+、Mg2+、Ba2+、Cu2+对该酶都具有激活作用且浓度至50 mmol/L时酶活力分别被激活至112%、127%、113%、128%、139%、199%、348%，然而十二烷基磺酸钠、抗坏血酸和草酸对该酶有强烈的抑制作用；Zn2+、甲醇、乙醇和异丙醇对该酶有一定的抑制作用。 |
| 关 键 词： | 黄瓜 过氧化物酶 分离纯化 性质 |
Isolation,Purification and Characterization of Peroxidase from Cucumber |
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| HU Rui-bin,LI Xing,WANG Hong-yang,WANG Jie,TANG Yun-ming. Isolation,Purification and Characterization of Peroxidase from Cucumber[J]. Food Science, 2014, 35(11): 168-173. DOI: 10.7506/spkx1002-6630-201411034 | |
| Authors: | HU Rui-bin LI Xing WANG Hong-yang WANG Jie TANG Yun-ming |
| Affiliation: | Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Sweet-Potato EngineeringResearch Center, School of Life Sciences, Southwest University, Chongqing 400715, China |
| Abstract: | Electrophoresis-purity peroxidase was extracted from fresh cucumber and purified through homogenization, extraction, ammonium sulfate precipitation, CM-Sepharose and Superdex-200 chromatography. The specific activity, recovery and purification fold of the peroxidase were 64 177.67 U/mg, 9.58% and 61.93, respectively. Molecular mass of this purified enzyme was 40.21 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), while native gel filtration confirmed a monomer form of 41.15 kD. Besides, the peroxidase, whose optimum temperature and pH were 60 ℃ and 6, respectively, was comparatively stable in the temperature range of 25-45 ℃ and pH range of 5-9. The purified peroxidase showed Km value of 53.79 mmol/L under definite conditions. In addition, its activity was scarcely affected by potassium thiocyanate (KSCN). The peroxidase was found to be activated by urea, K+, Mn2+, Ca2+, Mg2+, Ba2+ and Cu2+ by 12%, 27%, 13%, 28%, 39%, 99% and 248% at 50 mmol/L, respectively. However, the peroxidase activity was significantly inhibited by SDS, ascorbic acid and oxalic acid. Moreover, Zn2+, methanol, ethanol and isopropanol caused partial inhibitory effects on the peroxidase. |
| Keywords: | cucumber peroxidase isolation and purification characterization |
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