利用红外光谱与原子力显微技术揭示纳米级大豆肽结构及其与溶解性的关系
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摘要
以大豆分离蛋白为原料进行胃蛋酶水解,得到大豆肽。通过红外光谱、原子力显微镜对大豆蛋白和大豆肽的二级结构及微观结构进行分析,揭示结构与溶解性之间的关系。结果表明,与大豆分离蛋白相比,大豆肽二级结构主要以无规则卷曲为主,其α-螺旋和β-折叠结构含量低16.38%,而无序结构的β-转角和无规则卷曲含量高15.37%。大豆肽表面较平滑颗粒较分散,以单个分子状态存在,粒径为大豆分离蛋白的50%,在pH为4.0时,其溶解性为大豆分离蛋白的4.43倍。
Using the soy protein isolate after hydrolysis by pepsin as the substrate to obtain soybean peptides,the effects of secondary structure of soybean protein isolate and soybean peptide on the solubility of soy protein isolate were studied. The structure and microstructure of the two classes were analyzed by infrared spectroscopy and atomic force microscopy. The relationship between structure and function was analyzed by the functional analysis of the changes. The results showed that a-helix and β-sheet of the secondary structure of soybean peptide was 16.38% lower than that of soybean protein isolate,while the β-turn and random coil is about 15.37% higher than one.The surface of soybean peptide was smooth and the particle was dispersed,existing in single molecule state.The particle size was 50% of soybean protein isolate,and the solubility of soybean protein was 4.43 times of that of soybean protein isolate at pH4.0.
Using the soy protein isolate after hydrolysis by pepsin as the substrate to obtain soybean peptides,the effects of secondary structure of soybean protein isolate and soybean peptide on the solubility of soy protein isolate were studied. The structure and microstructure of the two classes were analyzed by infrared spectroscopy and atomic force microscopy. The relationship between structure and function was analyzed by the functional analysis of the changes. The results showed that a-helix and β-sheet of the secondary structure of soybean peptide was 16.38% lower than that of soybean protein isolate,while the β-turn and random coil is about 15.37% higher than one.The surface of soybean peptide was smooth and the particle was dispersed,existing in single molecule state.The particle size was 50% of soybean protein isolate,and the solubility of soybean protein was 4.43 times of that of soybean protein isolate at pH4.0.
引文
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[19]De-Bao Yuan,Wei Min,Xiao-Quan Yang,et al.An improved isolation method of soyβ-conglycinin subunitsand their characterization[J].J Am Oil Chem Soc,2010,87:997-1004.
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[24]Xiao-Zhong H U,Cheng Y Q,Fan J F,et al.Effects of drying method on physicochemical and functional properties of soy protein isolates[J].Journal of Food Processing and Preservation,2009,34(3):520-540.
[25]Bao X L,Lv Y,Yang B C,et al.A Study of the Soluble Complexes Formed during Calcium Binding by Soybean Protein Hydrolysates[J].Journal of Food Science,2008,73(3):117-121.
[26]刘永创,杨晓泉,郭健,等.等电点附近大豆分离蛋白乳化稳定性的研究[J].现代食品科技,2015(5):84-89.
[27]Ortiz S E M,An M C.Analysis of products,mechanisms of reaction,and some functional properties of soy protein hydrolysates[J].Journal of Oil and Fat Industries,2000,77(12):1293-1301.
[28]郭丽,王鹏,马松艳,等.枯草芽孢杆菌发酵大豆β-伴球蛋白产多肽的加工特性研究[J].食品工业,2013,34(4):18-21.
[29]Chen C,Chi Y J,Xu W.Comparisons on the Functional Properties and Antioxidant Activity of Spray-Dried and FreezeDried Egg White Protein Hydrolysate[J].Food and Bioprocess Technology,2011,5(6):1-11.
[30]Jbeily A C,Haubelt G,Myburgh J,et al.Results of an international ring test for the determination of water absorption capacity and physical properties of wheat dough using the Haubelt Flourgraph E 6(ICC standard no.179)[J].Molecular and General Genetics Mgg,2015,1(1):1-8.
[2]黄骊虹.大豆多肽的生理功能及应用(一)[J].食品科技,1999(30):50-51.
[3]Wiriyaumpaiwong S,Soponronnarit S,Prachayawarakorn S.Comparative study of heating processes for full-fat soybeans[J].Journal of Food Engineering,2004,65(3):371-382.
[4]Ortiz S E M,Wagner J R.Hydrolysates of native and modified soy protein isolates:structural characteristics,solubility and foaming properties[J].Food Research International,2002,35(6):511-518.
[5]Walsh D J,Cleary D,Mccarthy E,et al.Modification of the nitrogen solubility properties of soy protein isolate following proteolysis and transglutaminase cross-linking[J].Food Research International,2003,36(7):677-683.
[6]孙焕,张春红,陈海英.双酶复合改性增强大豆分离蛋白溶解性研究[J].粮食与油脂,2005(8):24-26.
[7]Zhang Z H,Yu-Fei A H.Determination of Interaction of Urea and Soybean Protein Isolate by Fourier Transform Infrared Spectroscopy[J].Soybean Science,2008,27(1):134-270.
[8]莫其逢,黄创高,田建民,等.原子力显微镜与表面形貌观察[J].广西物理,2007(2):46-49.
[9]Qi M,Hettiarachchy N S,Kalapathy U.Solubility and emulsifying properties of soy protein isolates modified by pancreatin[J].Journal of Food Science,1997,62(6):1110-1115.
[10]Baldauf N A,Rodriguez-Romo L A,Mannig A,et al.Effect of selective growth media on the differentiation of Salmonella enterica serovars by Fouriertransform mid-infrared spectroscopy[J].Journal of Microbiological Methods,2007 68(1):106-114.
[11]Dong A,Huang P,Caughey W S.Protein secondary structures in water from second-derivative amide I infrared spectral[J].Biochemistry,1990,29(13):3303-3308.
[12]Zhao J,Xiong Y L,McNear D H.Changes in structural characteristics of antioxidative soy protein hydrolysates resulting from scavenging of hydroxyl radicals[J].Journal of Food Science,2013,72(2):C152-C160.
[13]Alfredo Morales,Jozef L Kokini.Class transition of soy globulin using differential scanning calorimetry and mechanical spectrometry[J].Biotechnology Progress,1997,13(5):624-629.
[14]Subirade M,Kelly I,Gueguen J,et al.Molecular basis of film formation from a soybean protein:comparison between the conformation of glycinin in aqueous solution and in films[J].International Journal of Biological Macromolecules,1998,23(4):241-249.
[15]Jahaniaval F,Kakuda Y,Abraham V,et al.Soluble protein fractions from p H and heat treated sodium caseinate:physicochemical and functional properties[J].Food Research International,2000,33(8):637-647.
[16]Yu T,Zhao Y,Yu S.Progress on the EF-Hand proteins[J].Biophysics,2013(1):1-10.
[17]Wu W U,Hettiarachchy N S,Qi M.Hydrophobicity,solubility,and emulsifying properties of soy protein peptides prepared by papain modification and ultrafiltration[J].Journal of Oil and Fat Industries,1998,75(7):845-850.
[18]Akkermans C,Goot A J V D,Venema P,et al.Micrometersized fibrillar protein aggregates from soy glycin in and soy protein isolate[J].Journal of Agricultural and Food Chemistry,2007,55(24):9877-9882.
[19]De-Bao Yuan,Wei Min,Xiao-Quan Yang,et al.An improved isolation method of soyβ-conglycinin subunitsand their characterization[J].J Am Oil Chem Soc,2010,87:997-1004.
[20]Qi M,Hettiarachchy N S,Kalapathy U.Solubility and emulsifying properties of soy protein isolates modified by pancreatin[J].Journal of Food Science,1997,62(6):1110-1115.
[21]Tang C H&Wang C S.Formation and characterization of amyloid-like fibrils from soy b-conglycinin and glycinin[J].Journal of Agricultural and Food Chemistry,2010,58:11058-11066.
[22]刘骞.食品中蛋白质的功能(九)肉制品中蛋白质的功能特性[J].肉类研究,2009(12):58-66.
[23]石彦国,马永强,赵毅.大豆蛋白水解物物化特性的研究[J].中国粮油学报,2001,16(1):59-62.
[24]Xiao-Zhong H U,Cheng Y Q,Fan J F,et al.Effects of drying method on physicochemical and functional properties of soy protein isolates[J].Journal of Food Processing and Preservation,2009,34(3):520-540.
[25]Bao X L,Lv Y,Yang B C,et al.A Study of the Soluble Complexes Formed during Calcium Binding by Soybean Protein Hydrolysates[J].Journal of Food Science,2008,73(3):117-121.
[26]刘永创,杨晓泉,郭健,等.等电点附近大豆分离蛋白乳化稳定性的研究[J].现代食品科技,2015(5):84-89.
[27]Ortiz S E M,An M C.Analysis of products,mechanisms of reaction,and some functional properties of soy protein hydrolysates[J].Journal of Oil and Fat Industries,2000,77(12):1293-1301.
[28]郭丽,王鹏,马松艳,等.枯草芽孢杆菌发酵大豆β-伴球蛋白产多肽的加工特性研究[J].食品工业,2013,34(4):18-21.
[29]Chen C,Chi Y J,Xu W.Comparisons on the Functional Properties and Antioxidant Activity of Spray-Dried and FreezeDried Egg White Protein Hydrolysate[J].Food and Bioprocess Technology,2011,5(6):1-11.
[30]Jbeily A C,Haubelt G,Myburgh J,et al.Results of an international ring test for the determination of water absorption capacity and physical properties of wheat dough using the Haubelt Flourgraph E 6(ICC standard no.179)[J].Molecular and General Genetics Mgg,2015,1(1):1-8.