食品科学 ›› 2008, Vol. 29 ›› Issue (4): 41-45.

• 基础研究 •    下一篇

脂肪氧合酶催化亚油酸氧化与大豆蛋白相互作用过程中自由基迁移的电子顺磁共振研究(Ⅱ)自由基类型的确定

 黄友如, 华欲飞, 王雪峰, 顾建华, 陈义勇   

  1.  常熟理工学院生物与食品工程系; 江南大学食品学院; 常熟理工学院生物与食品工程系
  • 出版日期:2008-04-15 发布日期:2011-08-24

Electron Paramagnetic Resonance Spectroscopy (EPR) Study on Free Radical Transfer in Lipoxygenase I-B- Catalyzed Linoleic Acid-Soybean Protein Interaction (II) Determination of Free Radicals

HUANG  You-Ru, HUA  Yu-Fei, WANG  Xue-Feng, GU  Jian-Hua, CHEN  Yi-Yong   

  1. 1.Department of Biological Science and Food Engineering, Changshu Institute of Technology, Changshu 215500, China; 2.School of Food Science and Technology, Jiangnan University,
  • Online:2008-04-15 Published:2011-08-24

摘要: 模拟大豆蛋白制取过程,建立由亚油酸(LA)、脂肪氧合酶(LOX)和低脂质含量的大豆蛋白(LRSP,lipidreduced soybean protein)所组成的模拟体系并制备大豆蛋白,应用电子顺磁共振(EPR)比较研究了不同微波功率下的大豆蛋白EPR波谱,探讨了制备方法对自由基浓度和种类的影响,确定了四种类型的自由基。g值范围在2.0041~2.0054之间的碳自由基源于蛋白质肽链骨架α-碳原子或其侧链其他碳原子,其中场信号或称自由基浓度随微波功率的增加而降低。g值范围在2.019~2.028之间的硫自由基源于蛋白质中甲硫氨酸、半胱氨酸等含硫氨基酸残基侧链,它在室温下稳定,在不同微波功率下检测,在两个样品中均可观察到。羟基自由基.O H在两个样品中均可观察到,它源于自由基链反应和水分子,与前述大豆蛋白氧化性修饰的二酪氨酸形成有关。醛自由基一定源于大豆蛋白与LOX催化LA氧化产生的降解产物的酶促反应。它包含在中场碳自由基信号中,但稍稍偏向高场区。

关键词: 大豆蛋白, 脂肪氧合酶, 亚油酸, 自由基, 电子顺磁共振

Abstract: Soybean proteins were prepared from the model systems consisting of lipid-reduced soybean proteins, linoleic acid and lipoxygenase, and the EPR spectra of the proteins were investigated and compared with at different microwaves. The effects of preparation methods on the free radical concentration and species were examined. Four different type free radicals were identified as the damage of protein side chains or free radical chain reaction. The carbon radicals (g value range 2.0041 to 2.0054) formed at either on the backbone of the α-carbon or on the side chains of the proteins, while the free radical concentration or central singlet signal decreased with increased microwaves. Sulphur radicals (g value range 2.019 to 2.028) attributed to sulfur- containing side-chain of methionine or cysteine in the proteins were all detected at different microwaves, and were stable even at room temperature as observed in two samples. The radical ·OH was originated from free radical chain reaction and water molecules, and related to dityrosine formation from oxidative modification of soybean proteins examined in the previous study. Aldehyde radical must be originated from the enzymatic reactions which occurred between soybean proteins and the degradation product of linoleic acid oxidation catalyzed by lipoxygenase I-B (LOX). It was contained in the central carbon signal, but shifted in some sort to high magnetic field.

Key words: soybean proteins, lipoxygenase, linoleic acid, free radicals, EPR