两种天然抗氧化剂与鲢鱼肌球蛋白的相互作用研究

摘要/Abstract

摘要： 研究了两种天然抗氧化剂表没食子儿茶素没食子酸酯（Epigallocatechin gallate，EGCG）、白藜芦醇（Resveratrol，RE）与鲢鱼肌球蛋白之间的相互作用。采用荧光光谱技术探究了EGCG、RE与鲢鱼肌球蛋白分子间相互作用的结合情况；利用圆二色谱考察了EGCG、RE对鲢鱼肌球蛋白微观结构的影响。结果表明EGCG、RE与鲢鱼肌球蛋白之间发生相互作用会导致其荧光焠灭现象的发生，且荧光猝灭方式均为静态猝灭。热力学数据分析结果表明EGCG、RE与肌球蛋白分子的结合主要是氢键和范德华力作用力。利用 Stern -Volmer 方程处理实验数据，得到EGCG、RE与肌球蛋白在不同温度下的结合常数和结合位点数。圆二色谱结果表明EGCG，RE诱导肌球蛋白二级结构的变化，EGCG使肌球蛋白α-螺旋含量减少，RE使肌球蛋白α-螺旋含量增加。EGCG、RE都通过氢键和范德华力与鲢鱼肌球蛋白结合形成复合物，导致肌球蛋白发生荧光猝灭，并且改变了肌球蛋白构象。

关键词: 肌球蛋白, 表没食子儿茶素没食子酸酯, 白藜芦醇, 光谱技术, 相互作用

Abstract: The interaction between two kinds of natural antioxidant epigallocatechin gallate (EGCG), resveratrol (RE) and silver carp myosin was investigated. The binding constants and binding sites of the interaction between EGCG, RE and silver carp myosin explored by using fluorescence quenching method. Moreover, conformational changes and microstructure of silver carp myosin effected by EGCG and RE were determined through circular dichroism technique. The results showed that the interaction between EGCG, RE and silver carp myosin could cause the phenomenon of fluorescence quenching of myosin, and the style of fluorescence quenching was static quenching. Thermodynamic data analysis showed that the majority of intermolecular forces were hydrogen bonding and van Dewali interaction between EGCG, RE and myosin. By using the Stern-Volmer equation to analyze the experimental data, the value of binding constant and binding sites of EGCG and RE at different temperatures were obtained. The results of circular dichroism showed that EGCG and RE induced the change of secondary structure of myosin, EGCG reduced the content of myosin α-helix, but for RE it obtained completely opposite result compared with EGCG. Both EGCG and RE could interacted with silver carp myosin via hydrogen bonding and van Edward force to form complexes, which can lead to quenching of myosin fluorescence and change of the conformation of myosin.

Key words: Myosin, Epigallocatechin gallate, Resveratrol, Spectral technique, interaction

中图分类号:

TS254.1

黄渊杜红英. 两种天然抗氧化剂与鲢鱼肌球蛋白的相互作用研究[J]. 食品科学, 0, (): 0-0.

Yuan HUANG. Study on the Interaction between Two Kinds of Natural Antioxidants and Silver Carp Myosin[J]. FOOD SCIENCE, 0, (): 0-0.

参考文献

[1]农业部渔业局. 中国渔业年鉴[M]. 北京：中国农业出版社，2017: 21-36.
[2]王楚文, 柳云龙, 唐文静,等. 生物保鲜剂对鱼类保鲜的应用现状及其展望[J]. 安徽农业科学, 2015, 43(16):387-389. DOI：10.13989/j.cnki.0517-6611.2015.16.133
[3]陈义,汪小钢,宛晓春等.没食子儿茶素没食子酸酯制备新工艺的研究[J].安徽农业大学学报,2007,34(3):364-368. DOI:10.3969/j.issn.1672-352X.2007.03.012.
[4]李鹏，陈莉萍，黄琴等. EGCG对冷藏罗非鱼鱼片的保鲜[J]. 食品科技, 2014, 39(8) :118-122.DOI:10.13684/j.cnki.spkj.2014.08.027
[5]骆晓波. 表没食子儿茶素没食子酸酯纳米脂质体的制备及其对鳕鱼保鲜效果的研究[D]. 杭州：中国计量学院，2015：26-32.
[6]李先宽,李赫宇,李帅等.白藜芦醇研究进展[J].中草药,2016,47(14):2568-2578.DOI:10.7501/j.issn.0253-2670.2016.14.030.
[7]聂磊,叶敏仪,林静慧等.白藜芦醇和硝普钠对月季切花的保鲜效应[J].广州城市职业学院学报,2009,3(1):44-50.DOI:10.3969/j.issn.1674-0408.2009.01.010.
[8]金宏,惠伟,牛瑞雪等.白藜芦醇对砀山酥梨的保鲜效应[J].淮北师范大学学报（自然科学版）,2014, 35(01):51-55.DOI:10.3969/j.issn.2095-0691.2014.01.013.
[9]李强. 草鱼冷藏过程中肌肉蛋白质的组成和理化特性变化研究[D]. 长沙：长沙理工大学, 2013：11-21.
[10]朱道立,陈佩林,王康乐等.骨骼肌肌球蛋白Ⅱ的研究进展[J].生物学教学,2010,35(11):8-10.DOI:10.3969/j.issn.1004-7549.2010.11.004.
[11]PARK J A E W, LANIER T C. Scanning calorimetric behavior of tilapia myosin and actin due to processing of muscle and protein purification[J]. Journal of Food Science, 1989, 54(1): 49-51.DOI: 10.1111/j.1365-2621.1989.tb08564.x
[12]Lowry O H, Rosebrough N J, Farr A L, et al. Protein measurement with the Folin phenol reagent[J]. Journal of biological chemistry, 1951, 193(1): 265-275.
[13]YE J, FAN F, XU X, et al. Interactions of black and green tea polyphenols with whole milk[J]. Food Research International, 2013, 53(1):449-455. DOI: 10.1016/j.foodres.2013.05.033
[14]TAHA M, QUENTAL M V, CORREIA I, et al. Extraction and stability of bovine serum albumin (BSA) using cholinium-based Good's buffers ionic liquids[J]. Process Biochemistry, 2015, 50(7):1158-1166. DOI: 10.1016/j.procbio.2015.03.020
[15]王婧. 多酚类化合物与牛血清蛋白相互作用的研究[D]. 杭州：浙江工业大学, 2013：5-10.
[16]Bevington P R, Robinson D K, Blair J M, et al. Data reduction and error analysis for the physical sciences[J]. Computers in Physics, 1993, 7(4): 415-416. DOI: 10.1063/1.4823194
[17]Mandal P, Bardhan M, Ganguly T. A detailed spectroscopic study on the interaction of Rhodamine 6G with human hemoglobin[J]. Journal of Photochemistry and Photobiology B: Biology, 2010, 99(2): 78-86. DOI: 10.1016/j.jphotobiol.2010.02.009
[18]张淼,王道营,张牧焓等.一磷酸腺苷与肌动球蛋白相互作用的荧光光谱研究[J].食品科学,2016,37(17):32-37.DOI:10.7506/spkx1002-6630-201617006.
[19]林海彬,郑琳,林玉琴等.邻菲啰啉和氨三乙酸钴配合物与牛血清白蛋白相互作用的荧光分析[J].高等学校化学学报,2013,34(8):1818-1825.DOI:10.7503/cjcu20130098.
[20]Ross P D, Subramanian S. Thermodynamics of protein association reactions: forces contributing to stability[J]. Biochemistry, 1981, 20(11): 3096-3102.
[21]Klajnert B, Bryszewska M. Fluorescence studies on PAMAM dendrimers interactions with bovine serum albumin[J]. Bioelectrochemistry, 2002, 55(1): 33-35. DOI: 10.1016/S1567-5394(01)00170-0
[22]Azimi O, Emami Z, Salari H, et al. Probing the interaction of human serum albumin with norfloxacin in the presence of high-frequency electromagnetic fields: Fluorescence spectroscopy and circular dichroism investigations[J]. Molecules, 2011, 16(12): 9792-9818. DOI: 10.3390/molecules16129792.
[23]Chamani J, Tafrishi N, Momen-Heravi M. Characterization of the interaction between human lactoferrin and lomefloxacin at physiological condition: Multi-spectroscopic and modeling description[J]. Journal of Luminescence, 2010, 130(7):1160-1168. DOI: 10.1016/j.jlumin.2010.02.014
[24]尹燕霞,向本琼,佟丽.荧光光谱法在蛋白质研究中的应用[J].实验技术与管理, 2010, 27(2):33-36,40.DOI:10.3969/j.issn.1002-4956.2010.02.011.
[25]余晶梅. 荧光探针法和疏水相互作用层析法分析蛋白表面疏水性[D]. 杭州：浙江大学, 2014：28-37.
[26]王辰, 江连洲, 魏冬旭,等. 不同品种大豆分离蛋白结构与表面疏水性的关系[J]. 食品科学, 2012, 33(9):54-57.