柠檬黄与玉米醇溶蛋白的相互作用研究
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摘要
为了对玉米醇溶蛋白的改性研究及柠檬黄色素的安全使用评估提供理论依据,借助于荧光光谱法、紫外光谱法、全内反射-傅里叶红外光谱(ATR-FTIR)自去卷积计算及核磁共振氢谱研究了人工合成色素柠檬黄对玉米醇溶蛋白构象的影响。结果表明,柠檬黄对玉米醇溶蛋白有明显的荧光猝灭作用,猝灭机理较复杂。柠檬黄可以与玉米醇溶蛋白结合,分子间主要以疏水作用力结合,利用Stern-Volmer方程和Van't Hoff公式计算获得结合比为1∶1,结合常数K_a值较大。通过FTIR的自去卷积计算分析,这种结合反应导致玉米蛋白二级结构中α螺旋结构、β折叠和β转角均发生显著改变。核磁共振氢谱分析表明由于玉米醇溶蛋白具有两亲性,当溶剂为二甲基亚砜时,混合溶液为低极性环境,化学位移略向高磁场移动;当溶剂为重水D_2O时,化学位移明显地向低磁场移动,而且这种作用与反应时间长短无关。进一步说明在极性和低极性的环境下柠檬黄都会引起玉米醇溶蛋白构象的改变。
The effect of tartrazine on the zein conformation was studied by fluorescence spectroscopy, UV spectroscopy, ATR-FTIR self-deconvolution calculations and ~1H NMR spectroscopy, in order to provide a theoretical basis for the modification study of zein and the safe use assessment of tartrazine. The results showed that tartrazine had obvious fluorescence quenching effect on zein, and the quenching mechanism was complicated. Tartrazine can bind to zein, and the molecules were mainly combined by hydrophobic interaction. Using the Stern-Volmer equation and Van't Hoff formula, their binding ratio was calculated to be 1∶1 and the binding constant K_a value was larger. By self-deconvolution analysis of Fourier transform infrared spectroscopy, this binding reaction resulted in significant changes in the α-helix structure, β-folding and β-turn angle in the secondary structure of zein. Nuclear magnetic resonance spectroscopy indicated that due to the amphipathic nature of zein, when the solvent was dimethyl sulfoxide, the mixed solution was in a low-polarity environment, the chemical shift shifted slightly to a high magnetic field; when the solvent was heavy water D_2O, chemical shifts significantly shift to low magnetic fields, and this effect was independent of the length of reaction time. It was further illustrated that tartrazine can cause changes in the zein conformation in a polar and low polarity environment.
The effect of tartrazine on the zein conformation was studied by fluorescence spectroscopy, UV spectroscopy, ATR-FTIR self-deconvolution calculations and ~1H NMR spectroscopy, in order to provide a theoretical basis for the modification study of zein and the safe use assessment of tartrazine. The results showed that tartrazine had obvious fluorescence quenching effect on zein, and the quenching mechanism was complicated. Tartrazine can bind to zein, and the molecules were mainly combined by hydrophobic interaction. Using the Stern-Volmer equation and Van't Hoff formula, their binding ratio was calculated to be 1∶1 and the binding constant K_a value was larger. By self-deconvolution analysis of Fourier transform infrared spectroscopy, this binding reaction resulted in significant changes in the α-helix structure, β-folding and β-turn angle in the secondary structure of zein. Nuclear magnetic resonance spectroscopy indicated that due to the amphipathic nature of zein, when the solvent was dimethyl sulfoxide, the mixed solution was in a low-polarity environment, the chemical shift shifted slightly to a high magnetic field; when the solvent was heavy water D_2O, chemical shifts significantly shift to low magnetic fields, and this effect was independent of the length of reaction time. It was further illustrated that tartrazine can cause changes in the zein conformation in a polar and low polarity environment.
引文
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[2] AN B Z,WU X C,LI M J,et al..Hydrophobicity-modulating self-assembled morphologies of α-zein in aqueous ethanol [J].Int.J.Food Sci.Technol.,2016,51:2621-2629.
[3] 许辰琪,袁芳,高彦祥.玉米醇溶蛋白作为传递载体研究进展 [J].中国食品添加剂,2015(7):156-161.XU C Q,YUAN F,GAO Y X.Research progress of carrier material zein in delivery system [J].China Food Addit.,2015(7):156-161.(in Chinese)
[4] JOYE I J,DAVIDOV-PARDO G,LUDESCHER R D,et al..Fluorescence quenching study of resveratrol binding to zein and gliadin:towards a more rational approach to resveratrol encapsulation using water-insoluble proteins [J].Food Chem.,2015,185:261-267.
[5] SUN C X,DAI L,GAO Y X.Binary complex based on zein and propylene glycol alginate for delivery of quercetagetin [J].Biomacromolecules,2016,17(12):3973-3985.
[6] GOMEZ-ESTACA J,BALAGUER M P,GAVARA R,et al..Formation of zein nanoparticles by electrohydrodynamic atomization:effect of the main processing variables and suitability for encapsulating the food coloring and active ingredient curcumin [J].Food Hydrocoll.,2012,28(1):82-91.
[7] 王永辉,杨晓泉,王金梅,等.蛋白水解物及多糖负载姜黄素制备纳米颗粒及其稳定性 [J].农业工程学报,2015,31(10):296-302.WANG Y H,YANG X Q,WANG J M,et al..Preparation of curcumin nanoparticles by protein hydrolysates and polysaccharids and its stabilization [J].Trans.Chin.Soc.Agric.Eng.,2015,31(10):296-302.(in Chinese)
[8] WANG H L,HAO L L,WANG P,et al..Release kinetics and antibacterial activity of curcumin loaded zein fibers [J].Food Hydrocoll.,2017,63:437-446.
[9] YILMAZ A,BOZKURT F,CICEK P K,et al..A novel antifungal surface-coating application to limit postharvest decay on coated apples:molecular,thermal and morphological properties of electrospun zein-nanofiber mats loaded with curcumin [J].Food Sci.Emerg.Technol.,2016,37:74-83.
[10] 魏莹,吴之翔.简述着色剂在调味品、食品中的应用 [J].中国调味品,2014,39(5):126-128.WEI Y,WU Z X.Brief description of application of colorants in condiments and foods [J].China Cond.,2014,39(5):126-128.(in Chinese)
[11] 陈启振,曾勇明,林惠真,等.表面增强拉曼光谱在食品人工合成色素的现场快速筛查中的应用 [J].厦门大学学报(自然科学版),2016,55(5):754-759.CHEN Q Z,ZENG Y M,LIN H Z,et al..Developing on-site,quick screening platform for artificial pigments in food using surface-enhanced Raman spectroscopy [J].J.Xiamen Univ.(Nat.Sci.),2016,55(5):754-759.(in Chinese)
[12] SAXENA B,SHARMA S.Food color induced hepatotoxicity in Swiss albino rats,Rattus norvegicus [J].Toxicol.Int.,2015,22(1):152-157.
[13] AL-SHABIB N A,KHAN J M,ALSENAIDY M A,et al..Unveiling the stimulatory effects of tartrazine on human and bovine serum albumin fibrillogenesis:spectroscopic and microscopic study [J].Spectrochim.Acta Part A:Mol.Biomol.Spectrosc.,2018,191:116-124.
[14] AL-SHABIB N A,KHAN J M,KHAN M S,et al..Synthetic food additive dye “Tartrazine” triggers amorphous aggregation in cationic myoglobin [J].Int.J.Biol.Macromol.,2017,98:277-286.
[15] PANJA S,HALDER M.Exploration of electrostatic interaction in the hydrophobic pocket of lysozyme:importance of ligand-induced perturbation of the secondary structure on the mode of binding of exogenous ligand and possible consequences [J].J.Photochem.Photobiol.B Biol.,2016,161:253-265.
[16] 赵芳,黄超锋,梁慧,等.大黄酸锌配合物与牛血清白蛋白相互作用的研究 [J].分析化学,2011,39(3):401-404.ZHAO F,HUANG C F,LIANG H,et al..Interaction of Zn(Ⅱ) complex of rhein with bovine serum albumin [J].Chin.J.Anal.Chem.,2011,39(3):401-404.(in Chinese)
[17] BYLER D M,SUSI H.Examination of the secondary structure of proteins by deconvolved FTIR spectra [J].Biopolymers,1986,25(3):469-487.
[18] 卢雁,张玮玮,王公轲.FTIR用于变性蛋白质二级结构的研究进展 [J].光谱学与光谱分析,2008,28(1):88-93.LU Y,ZHANG W W,WANG G K.Progress in study of secondary structure of denaturized protein by FTIR [J].Spectrosc.Spectr.Anal.,2008,28(1):88-93.(in Chinese)
[19] 刘保生,李彤彤,张秋菊,等.不同光谱法用于药物与蛋白相互作用的比较研究 [J].发光学报,2016,37(7):866-872.LIU B S,LI T T,ZHANG Q J,et al..Multi-spectrophotometric methods for the investigation of the interactions of bovine serum albumin and drugs [J].Chin.J.Lumin.,2016,37(7):866-872.(in Chinese)
[20] 张颖,钟莉,杜静,等.多酚与蛋白质相互作用的荧光内滤效应校正方法的选择 [J].光谱学与光谱分析,2014,34(1):116-121.ZHANG Y,ZHONG L,DU J,et al..Assessment of inner filter effect corrections in fluorimetry of the interaction between polyphenols and proteins [J].Spectrosc.Spectr.Anal.,2014,34(1):116-121.(in Chinese)
[21] 许金钩,王尊本.荧光分析法 [M].3版.北京:科学出版社,2006.XU J G,WANG Z B.Fluorimetry [M].3rd ed.Beijing:Science Press,2006.(in Chinese)
[22] LI J F,LI J Z,YONG J,et al..Spectroscopic analysis and molecular modeling on the interaction of jatrorrhizine with human serum albumin (HSA) [J].Spectrochim.Acta Part A:Mol.Biomol.Spectrosc.,2014,118:48-54.