碱性电解水对籽瓜种仁蛋白质提取效果的影响
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摘要
传统的蛋白提取方法多以碱溶酸沉法为主,由于在提取过程中需要使用强酸或强碱调节pH值,蛋白质的结构与功能会产生不同程度的破坏,并使水体生态环境恶化。为探寻一种新型提取方法,本研究以电解水为提取溶剂,与传统碱溶酸沉法提取籽瓜种仁蛋白的效果进行对比。结果表明:在相同的pH值(pH 9.5)条件下,电解水提取的籽瓜种仁蛋白质(electrolyzed water extracted seed kernel protein,EWP)相比于超纯水提取的籽瓜种仁蛋白质(ultrapure water extracted seed kernel protein,UWP)无论是在提取率还是蛋白质的溶解性、乳化性和起泡性方面都有显著提高(P<0.05)。在必需氨基酸组成方面,EWP中半胱氨酸、缬氨酸、异亮氨酸、亮氨酸的含量相比于UWP有显著提高(P<0.05)。同时,EWP的二级结构更为有序,其α-螺旋和β-折叠有序结构比例分别为60.2%和2.3%,而β-转角和无规卷曲的无序结构比例则分别为35.8%和1.7%。由此可知,相比于超纯水提取法,碱性电解水提取法不仅可以提高籽瓜种仁蛋白质的提取率,而且对其结构和功能破坏较小,是一种高效、绿色、环保的提取方法。
Traditionally, alkali dissolution and acid precipitation is used for protein extraction. The structure and function of proteins are damaged due to the use of a strong acid or alkali to adjust the pH during the extraction process and, at the same time, the aqueous environment is deterioratedapplication. In order to develop a new extraction method, in the present study,electrolyzed water was used as a solvent to extract seed kernel protein from Citrullus lanatus var. megalaspermus and the extraction ef?ciency was compared with that of the alkali dissolution and acid precipitation method(using ultrapure water as extraction solvent). The results revealed that at pH 9.5, the yield, solubility, and emulsifying and foaming properties of electrolyzed water extracted seed kernel protein(EWP) were signi?cantly superior to those of ultrapure water extracted seed kernel protein(UWP)(P < 0.05). In the case of essential amino acid composition, cysteine, proline, isoleucine and leucine in EWP were signi?cantly more abundant than in UWP(P < 0.05). Moreover, andwith the secondary structure of EWP was more ordered; the relative contents α-helix and β-sheet(ordered) were 60.2% and 2.3%, and the relative contents of β-turn and random coil(disordered) were 35.8% and 1.7%. In conclusion, compared with ultrapure water extraction,electrolyzed water extraction could not only increase the extraction yield of protein from seed kernels of Citrullus lanatus var. megalaspermus, but also cause less damage to protein structure and function and was an ef?cient and environmentally friendly extraction method.
Traditionally, alkali dissolution and acid precipitation is used for protein extraction. The structure and function of proteins are damaged due to the use of a strong acid or alkali to adjust the pH during the extraction process and, at the same time, the aqueous environment is deterioratedapplication. In order to develop a new extraction method, in the present study,electrolyzed water was used as a solvent to extract seed kernel protein from Citrullus lanatus var. megalaspermus and the extraction ef?ciency was compared with that of the alkali dissolution and acid precipitation method(using ultrapure water as extraction solvent). The results revealed that at pH 9.5, the yield, solubility, and emulsifying and foaming properties of electrolyzed water extracted seed kernel protein(EWP) were signi?cantly superior to those of ultrapure water extracted seed kernel protein(UWP)(P < 0.05). In the case of essential amino acid composition, cysteine, proline, isoleucine and leucine in EWP were signi?cantly more abundant than in UWP(P < 0.05). Moreover, andwith the secondary structure of EWP was more ordered; the relative contents α-helix and β-sheet(ordered) were 60.2% and 2.3%, and the relative contents of β-turn and random coil(disordered) were 35.8% and 1.7%. In conclusion, compared with ultrapure water extraction,electrolyzed water extraction could not only increase the extraction yield of protein from seed kernels of Citrullus lanatus var. megalaspermus, but also cause less damage to protein structure and function and was an ef?cient and environmentally friendly extraction method.
引文
[1]曹承旭,马越,赵晓燕,等.麦芽糊精质量分数对籽瓜种子蛋白粉特性的影响[J].食品科技,2017,42(10):90-96.DOI:10.13684/j.cnki.spkj.2017.10.019.
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[9]TAMAKI S,BUI V N,NGO L H,et al.Virucidal effect of acidic electrolyzed water and neutral electrolyzed water on avian influenza viruses[J].Archives of Virology,2014,159(3):405-412.DOI:10.1007/s00705-013-1840-2.
[10]W A T A N A B E M,M A E D A I,K O Y A M A M,e t a l.Simultaneousrecoveryand purificationof riceproteinandphosphoru scompoundsfrom full-fatanddefattedricebranwith organicsolventfreeprocess[J].Journal of Fermentation and Bioengineering,2015,119(2):206-211.DOI:10.1016/j.jbiosc.2014.07.009.
[11]TOGE Y,MIYASHITA K.Lipid extraction with electrolyzed cathode water from marine products[J].Journal of Oleo Science,2003,52(1):1-6.DOI:10.5650/jos.52.1.
[12]NABOK M V,PLUTAHIN G A.Study and development of new technology for baking wheat bread by using electro-activated water as replacement to ordinary water[J].Scientific Communications of the Kuban State Agrarian University,2005,2005:23.
[13]WU Wei,ZHANG Caimeng,KONG Xiangzhen,et al.Oxidative modification of soy protein by peroxyl radicals[J].Food Chemistry,2009,116(1):295-301.DOI:10.1016/j.foodchem.2009.02.049.
[14]KATO A,NAKAI S.Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins[J].Biochimica et Biophysica Acta,1980,624(1):13-20.DOI:10.1016/0005-2795(80)90220-2.
[15]BEVERIDGE T,TOMA S J,NAKAI S.Determination of SH-and S-S groups in some food proteins using Ellman’s Reagent[J].Journal of Food Science,1974,39(1):49-51.DOI:10.1111/j.1365-2621.1974.tb00984.x.
[16]BERA M B,MUKHERJEE R K.Solubility,emulsifying,and foaming properties of rice bran protein concentrates[J].Journal of Food Science,1989,54(1):142-145.DOI:10.1111/j.1365-2621.1989.tb08587.x.
[17]PEARCE K N,KINSELLA J E.Emulsifying properties of proteins:evaluation of a turbidimetric technique[J].Journal of Agricultural and Food Chemistry,1978,26(3):716-723.DOI:10.1021/jf60217a041.
[18]BANDYOPADHYAY K,MISRA G,GHOSH S.Preparation and characterisation of protein hydrolysates from Indian defatted rice bran meal[J].Journal of Oleo Science,2008,57(1):47-52.DOI:10.5650/jos.57.47.
[19]LI Y,JONGBERG S,ANDERSEN M L,et al.Quinone-induced protein modifications:kinetic preference for reaction of 1,2-benzoquinones with thiol groups in proteins[J].Free Radical Biology and Medicine,2016,97:148-157.DOI:10.1016/j.freeradbiomed.2016.05.019.
[20]VENKATACHALAM M,ROUX K H,SATHE S K.Biochemical characterization of soluble proteins in pecan Carya illinoinensis(Wangenh.)K.Koch[J].Journal of Agricultural and Food Chemistry,2008,56(17):8103-8110.DOI:10.1021/jf801268k.
[21]PARK C,RAINES R T.Adjacent cysteine residues as a redox switch[J].Protein Engineering Design and Selection,2001,14(11):939-942.DOI:10.1093/protein/14.11.939.
[22]BOATRIGHT W L,HETTIARACHCHY N S.Effect of lipids on soy protein isolate solubility[J].Journal of the American Oil Chemists’Society,1995,72(12):1439-1444.DOI:10.1007/bf02577835.
[23]王国珍.新型蛋白质氧化折叠助剂的分子设计和复性方法研究[D].天津:天津大学,2011:3.
[24]张辉.离子强度对大豆分离蛋白结构及表面疏水性的影响[J].食品工业科技,2016,37(8):145-149;154.DOI:10.13386/j.issn1002-0306.2016.08.021.
[25]刘伯业.小麦蛋白低苦味肽的制备及其脱苦机理研究[D].无锡:江南大学,2017:4.
[26]WHITMORE L,WALLACE B A.Protein secondary structure analyses from circular dichroism spectroscopy:methods and reference databases[J].Biopolymers,2008,89(5):392-400.DOI:10.1002/bip.20853.
[27]CARBONARO M,NUCARA A.Secondary structure of food proteins by Fourier transform spectroscopy in the mid-infrared region[J].Amino Acids,2010,38(3):679-690.DOI:10.1007/s00726-009-0274-3.
[28]沈琼,黄滨,邵嘉亮,等.运用圆二色谱研究酶与化合物相互作用的机理[J].中山大学学报(自然科学版),2006,45(4):62-64.DOI:10.3321/j.issn:0529-6579.2006.04.015.
[29]JIANG J,CHEN J,XIONG Youling L..Structural and emulsifying properties of soy protein isolate subjected to acid and alkaline pH-shifting processes[J].Journal of Agricultural and Food Chemistry,2009,57(16):7576-7583.DOI:10.1021/jf901585n.
[30]魏晓芳,刘会洲.热变性对蛋白质结构及泡沫行为的影响[J].化工冶金,2000,21(4):379-383.DOI:10.3321/j.issn:1009-606X.2000.04.010.
[31]王一博.高起泡性蛋清液制备与应用研究[D].武汉:华中农业大学,2016:33.
[2]陆晓婷,张超,张晖,等.黑籽瓜种子蛋白酶解工艺及抗氧化性研究[J].食品科技,2015,40(5):73-78.DOI:10.13684/j.cnki.spkj.2015.05.015.
[3]张超,郭晓飞,李武,等.籽瓜种子蛋白质组成及其提取工艺优化的研究[J].中国粮油学报,2013,28(9):119-123.DOI:10.3969/j.issn.1003-0174.2013.09.023.
[4]SHAPOSHNIK V A,KESORE K.An early history of electrodialysis with permselective membranes[J].Journal of Membrane Science,1997,136(1/2):35-39.DOI:10.1016/S0376-7388(97)00149-X.
[5]陶国枢,胡亚卓,韩志涛,等.碱性电解水对血脂影响的实验研究[J].中国老年保健医学杂志,2003,1(2):18-20.DOI:10.3969/j.issn.1672-4860-B.2003.02.005.
[6]王晓娟,冯浩,李志义.电解水预处理对玉米秸秆和柳枝稷酶解产糖的研究[J].高校化学工程学报,2012,26(1):169-174.
[7]裴海生,孙君社.强电解水及其在食品工业中的应用[J].中国食品学报,2011,11(9):268-271.DOI:10.3969/j.issn.1009-7848.2011.09.029.
[8]KOBAYASHI K,HARA Y,KATSUBE T.Extraction of mulberry leaf components using an ethanol solution with electrolyzed water[J].Nippon Shokuhin Kogyo Gakkaishi,2006,53(10):548-550.DOI:10.3136/nskkk.53.548.
[9]TAMAKI S,BUI V N,NGO L H,et al.Virucidal effect of acidic electrolyzed water and neutral electrolyzed water on avian influenza viruses[J].Archives of Virology,2014,159(3):405-412.DOI:10.1007/s00705-013-1840-2.
[10]W A T A N A B E M,M A E D A I,K O Y A M A M,e t a l.Simultaneousrecoveryand purificationof riceproteinandphosphoru scompoundsfrom full-fatanddefattedricebranwith organicsolventfreeprocess[J].Journal of Fermentation and Bioengineering,2015,119(2):206-211.DOI:10.1016/j.jbiosc.2014.07.009.
[11]TOGE Y,MIYASHITA K.Lipid extraction with electrolyzed cathode water from marine products[J].Journal of Oleo Science,2003,52(1):1-6.DOI:10.5650/jos.52.1.
[12]NABOK M V,PLUTAHIN G A.Study and development of new technology for baking wheat bread by using electro-activated water as replacement to ordinary water[J].Scientific Communications of the Kuban State Agrarian University,2005,2005:23.
[13]WU Wei,ZHANG Caimeng,KONG Xiangzhen,et al.Oxidative modification of soy protein by peroxyl radicals[J].Food Chemistry,2009,116(1):295-301.DOI:10.1016/j.foodchem.2009.02.049.
[14]KATO A,NAKAI S.Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins[J].Biochimica et Biophysica Acta,1980,624(1):13-20.DOI:10.1016/0005-2795(80)90220-2.
[15]BEVERIDGE T,TOMA S J,NAKAI S.Determination of SH-and S-S groups in some food proteins using Ellman’s Reagent[J].Journal of Food Science,1974,39(1):49-51.DOI:10.1111/j.1365-2621.1974.tb00984.x.
[16]BERA M B,MUKHERJEE R K.Solubility,emulsifying,and foaming properties of rice bran protein concentrates[J].Journal of Food Science,1989,54(1):142-145.DOI:10.1111/j.1365-2621.1989.tb08587.x.
[17]PEARCE K N,KINSELLA J E.Emulsifying properties of proteins:evaluation of a turbidimetric technique[J].Journal of Agricultural and Food Chemistry,1978,26(3):716-723.DOI:10.1021/jf60217a041.
[18]BANDYOPADHYAY K,MISRA G,GHOSH S.Preparation and characterisation of protein hydrolysates from Indian defatted rice bran meal[J].Journal of Oleo Science,2008,57(1):47-52.DOI:10.5650/jos.57.47.
[19]LI Y,JONGBERG S,ANDERSEN M L,et al.Quinone-induced protein modifications:kinetic preference for reaction of 1,2-benzoquinones with thiol groups in proteins[J].Free Radical Biology and Medicine,2016,97:148-157.DOI:10.1016/j.freeradbiomed.2016.05.019.
[20]VENKATACHALAM M,ROUX K H,SATHE S K.Biochemical characterization of soluble proteins in pecan Carya illinoinensis(Wangenh.)K.Koch[J].Journal of Agricultural and Food Chemistry,2008,56(17):8103-8110.DOI:10.1021/jf801268k.
[21]PARK C,RAINES R T.Adjacent cysteine residues as a redox switch[J].Protein Engineering Design and Selection,2001,14(11):939-942.DOI:10.1093/protein/14.11.939.
[22]BOATRIGHT W L,HETTIARACHCHY N S.Effect of lipids on soy protein isolate solubility[J].Journal of the American Oil Chemists’Society,1995,72(12):1439-1444.DOI:10.1007/bf02577835.
[23]王国珍.新型蛋白质氧化折叠助剂的分子设计和复性方法研究[D].天津:天津大学,2011:3.
[24]张辉.离子强度对大豆分离蛋白结构及表面疏水性的影响[J].食品工业科技,2016,37(8):145-149;154.DOI:10.13386/j.issn1002-0306.2016.08.021.
[25]刘伯业.小麦蛋白低苦味肽的制备及其脱苦机理研究[D].无锡:江南大学,2017:4.
[26]WHITMORE L,WALLACE B A.Protein secondary structure analyses from circular dichroism spectroscopy:methods and reference databases[J].Biopolymers,2008,89(5):392-400.DOI:10.1002/bip.20853.
[27]CARBONARO M,NUCARA A.Secondary structure of food proteins by Fourier transform spectroscopy in the mid-infrared region[J].Amino Acids,2010,38(3):679-690.DOI:10.1007/s00726-009-0274-3.
[28]沈琼,黄滨,邵嘉亮,等.运用圆二色谱研究酶与化合物相互作用的机理[J].中山大学学报(自然科学版),2006,45(4):62-64.DOI:10.3321/j.issn:0529-6579.2006.04.015.
[29]JIANG J,CHEN J,XIONG Youling L..Structural and emulsifying properties of soy protein isolate subjected to acid and alkaline pH-shifting processes[J].Journal of Agricultural and Food Chemistry,2009,57(16):7576-7583.DOI:10.1021/jf901585n.
[30]魏晓芳,刘会洲.热变性对蛋白质结构及泡沫行为的影响[J].化工冶金,2000,21(4):379-383.DOI:10.3321/j.issn:1009-606X.2000.04.010.
[31]王一博.高起泡性蛋清液制备与应用研究[D].武汉:华中农业大学,2016:33.