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当前位置: 首页 > 文章 > 黑豆脂氧合酶的制备及其酶学活性的鉴定 食品科学 2016,37 (9) 155-159
Position: Home > Articles > Preparation and Identification of a Black Soybean Lipoxygenase FOOD SCIENCE 2016,37 (9) 155-159

黑豆脂氧合酶的制备及其酶学活性的鉴定

作  者：
李茹;崔晓东;李玉英;李晨;王转花
单  位：
山西大学生命科学学院;山西大学化学生物学与分子工程教育部重点实验室
关键词：
黑豆;脂氧合酶;β-葡萄糖苷酶活性;花色苷;花色素
摘  要：
从黑豆种子中分离纯化1种黑豆脂氧合酶——bs LOX,对其酶学活性及降解花色苷等性质进行初步研究。采用缓冲液抽提、硫酸铵沉淀、透析、阳离子交换层析及凝胶层析等方法纯化出具有β-葡萄糖苷酶活性的bs LOX;分别以亚油酸钠和对硝基苯基-β-D-吡喃葡萄糖苷(p-nitrophenylβ-D-glucopyranoside,p NPG)为反应底物鉴定bs LOX的脂氧合酶活性和β-葡萄糖苷酶活性。从黑豆种子中纯化获得的bs LOX在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electropheresis,SDS-PAGE)图谱显示单一条带,分子质量约为95 k D。酶活性分析表明,纯化的bs LOX以亚油酸钠为底物时,酶活力为13 U/m L;当以p NPG为底物在405 nm波长处检测到了对硝基苯酚的生成,从而推测bs LOX具有β-葡萄糖苷酶活性。高效液相色谱实验结果表明bs LOX可以降解花色苷(矢车菊素-3-O-葡萄糖苷、矮牵牛素-3-O-葡萄糖苷)为花色素(矢车菊素、矮牵牛素)。由于降解作用不完全,继而发现花色素能抑制bs LOX的酶活性。bs LOX是黑豆中的一种兼具β-葡萄糖苷酶功能的蛋白质,而花色素可能是天然的bs LOX抑制剂。
译  名：
Preparation and Identification of a Black Soybean Lipoxygenase
作  者：
LI Ru;CUI Xiaodong;LI Yuying;LI Chen;WANG Zhuanhua;College of Life Science,Shanxi University;Key Laboratory of Chemical Biology and Molecular Engineering,Ministry of Education,Shanxi University;
关键词：
black soybean;;lipoxygenase;;β-glucosidase activity;;anthocyanin;;anthocyanidins
摘  要：
Objective: To extract and purify black soybean lipoxygenase(bs LOX) containing β-glucosidase activity and investigate its anthocyanins-degrading properties and enzymatic activity. Methods: Purification of bs LOX was carried out by buffer extraction, ammonium sulfate precipitation, dialysis, cation exchange chromatography and gel filtration chromatography. The activities of lipoxygenase and β-glucosidase in bs LOX were analyzed by using sodium linoleate and p-nitrophenyl β-D-glucopyranoside(p NPG) as substrates, respectively. An LOX from black soybean seeds was obtained, which showed a single band in SDS-PAGE, with approximate molecular weight of 95 k D. The purified protein had a lipoxygenase activity of approximately 13 U/m L, and could produce nitro phenol using p NPG as substrate as detected at 405 nm, suggesting that bs LOX had β-glucosidase activity. HPLC results showed that bs LOX could only degrade part of anthocyanins(cyanidin-3-O-glucoside and petunidin-3-O-glucoside) into aglycone anthocyanidins(cyanidin and petunidin) and glucose; in turn, the residual anthocyanidins could inhibit the activity of bs LOX. Conclusion: LOX in black soybean may be a kind of protein with a variety of enzymatic functions, while anthocyanidins may be a natural bs LOX inhibitor.

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