Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (22): 4894-4905.doi: 10.3864/j.issn.0578-1752.2021.22.015

• FOOD SCIENCE AND ENGINEERING • Previous Articles     Next Articles

Regulation Effects of Ultrasound on the Structure and Emulsification Properties of Pea Protein Isolate

LI ZhaoRui1(),HAN XinRui1,FAN Xin1,HUANG JunRong1,CAO YunGang1(),XIONG YouLing2()   

  1. 1School of Food and Biological Engineering, Shaanxi University of Science & Technology, Xi’an 710021, China
    2Department of Animal and Food Sciences, University of Kentucky, Lexington 40546, KY, United States
  • Received:2021-04-11 Accepted:2021-07-27 Online:2021-11-16 Published:2021-11-19
  • Contact: YunGang CAO,YouLing XIONG E-mail:1762903310@qq.com;caoyungang@sust.edu.cn;ylxiong@uky.edu

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Abstract:

【Objective】 This study was designed to investigate the effects of ultrasonic treatment on the structure and physicochemical properties of pea protein isolate (PPI), and to explore the regulation mechanism of ultrasonic treatment on the emulsifying characteristics of PPI, so as to provide the theoretical basis for the application of PPI as natural emulsions or related products in the food field. 【Method】 The ultrasonic-pea protein isolate (U-PPI) was prepared by ultrasonic wave with frequency of 20 kHz and power of 600 W under different times (0, 20, 30, 40 and 60 min), and then U-PPI emulsions were prepared by high pressure homogenization. The effects of ultrasonic treatment on the physicochemical properties of PPI were investigated by the tests of free amino group, total sulfhydryl group, particle size, solubility and SDS-PAGE. The changes in secondary and tertiary structure of U-PPI were analyzed by circular dichroism and intrinsic tryptophan fluorescence, respectively. The emulsifying ability and emulsion stability of U-PPI were characterized by emulsion activity index, particle size, protein distribution at the interface of emulsion, Zeta potential and apparent viscosity. The microstructure of the emulsions was observed by using laser confocal fluorescence microscopy. 【Result】 Ultrasonic treatment significantly modified the structure of PPI, i.e., a short time (30 to 40 min) ultrasonic treatment significantly reduced the α-helix content and improved the β-sheet content of PPI, which made the structure of the PPI more flexible and more hydrophobic groups exposed to the interface. Simultaneously, the depolymerization effect of ultrasonic also caused the decrease of the average particle size and the significant increase of the solubility of PPI. Therefore, the modification of PPI by a short time ultrasonic treatment was conducive to the formation of a dense and stable protein film at the oil/water interface, which effectively improved the emulsifying activity of PPI and the stability of the emulsions, and the microstructure of the emulsions also showed that the particle sizes of emulsions stabilized with U-PPI were smaller, and the droplet distribution was more uniform. However, a long time (60 min) ultrasonic treatment caused the hydrophobic repolymerization and insolubility of PPI, which was not conducive to the adsorption and rearrangement of PPI in the oil/water interface and thus reduced the emulsifying activity and emulsion stability. 【Conclusion】 The cavitation effect and mechanical effect produced by a short time (30, 40 min) ultrasonic treatment had significant depolymerization effect on PPI, which promoted the protein structure more flexible and was beneficial to the adsorption and rearrangement of PPI at the oil/water interface, and thus significantly improved the emulsifying properties of PPI.

Key words: ultrasonic modification, pea protein isolate, structural modification, emulsion stability

Fig. 1

Effect of different ultrasonic treatment time on free amino group and total sulfhydryl group contents of PPI Con: Unultrasonic treatment. Different letters a-e or A-D indicate the same indexes are significantly different among different treatments (P<0.05), respectively. The same as below"

Fig. 2

Effect of different ultrasonic treatment time on the secondary structure of PPI"

Fig. 3

Effects of different ultrasonic treatment time on the intrinsic tryptophan fluorescence of PPI"

Fig. 4

Effects of different ultrasonic treatment time on the mean particle size and solubility of PPI"

Fig. 5

Effects of different ultrasonic treatment time on the protein subunits of PPI"

Fig. 6

Effects of different ultrasonic treatment time on the emulsifying activity of PPI"

Fig. 7

Effects of different ultrasonic treatment time on the particle size distribution and Zeta potential of PPI emulsions"

Table 1

Effect of different ultrasonic treatment time on the protein distribution at the interface of PPI emulsions"

时间
Time (min)		 水相蛋白
Aqueous protein (%)		 界面蛋白
Interface protein (%)		 分配系数
Partition coefficient
Con 38.0±0.0e 62.0±0.0e 14.75
20 29.7±1.2b 70.3±2.1b 21.63
30 27.3±1.2a 72.7±1.5a 23.95
40 31.7±0.6c 68.3±2.1c 19.51
60 33.0±0.0d 67.0±0.0d 18.19

Fig. 8

Effects of different ultrasonic treatment time on the apparent viscosity of PPI emulsions"

Fig. 9

Laser confocal micrograph of emulsions stabilized with PPI treated with different ultrasonic time Nile red and Nile blue are used to stain oil droplets (red) and water phase proteins (blue), respectively; The figures above show the distributions of individual oil droplets and the figures below show the distributions of oil droplets in the aqueous phase"

[1] NESTERENKO A, ALRIC I, SILVESTRE F, DURRIEU V. Influence of soy protein's structural modifications on their microencapsulation properties: α-Tocopherol microparticle preparation. Food Research International, 2012, 48(2): 387-396.
doi: 10.1016/j.foodres.2012.04.023
[2] 李慧, 王琪, 付鑫, 王鑫烁, 周泉城. 豌豆蛋白挤出物制备抗氧化肽工艺优化及其分离. 食品工业科技, 2018, 39(6): 165-169.
LI H, WANG Q, FU X, WANG X S, ZHOU Q C. Optimization of preparation technology and separation of antioxidant peptides from pea protein extrudate. Science and Technology of Food Industry, 2018, 39(6): 165-169. (in Chinese)
[3] HAFIZ R S, PETER A. W, MIAN K S, SHABBAR A, HAMID M, KINGSLEY G M, WASEEM S, FANG Z. Current progress in the utilization of native and modified legume proteins as emulsifiers and encapsulants-A review. Food Hydrocolloids, 2018, 76: 2-16.
doi: 10.1016/j.foodhyd.2017.01.002
[4] 孙英杰. 超声波处理对大豆分离蛋白结构和功能性质影响研究[D]. 哈尔滨: 东北农业大学, 2014.
SUN Y J. Study on the effect of ultrasonic treatments on the structure and functional properties of SPI[D]. Harbin: Northeast Agricultural University, 2014. (in Chinese)
[5] PENG W W, KONG X Z, CHEN Y M, ZHANG C M, YANG Y X, HUA Y F. Effects of heat treatment on the emulsifying properties of pea proteins. Food Hydrocolloids, 2016, 52: 301-310.
doi: 10.1016/j.foodhyd.2015.06.025
[6] YIN S W, TANG C H, WEN Q B, YANG X Q, LI L. Functional properties and in vitro trypsin digestibility of red kidney bean (Phaseolus vulgaris L.) protein isolate: Effect of high-pressure treatment. Food Chemistry, 2008, 110(4): 938-945.
doi: 10.1016/j.foodchem.2008.02.090
[7] SILVA J L, WEBER G. Pressure stability of proteins. Annual Review of Physical Chemistry, 1993, 44: 89-113.
doi: 10.1146/physchem.1993.44.issue-1
[8] SHEN L, TANG C H. Microfluidization as a potential technique to modify surface properties of soy protein isolate. Food Research International, 2012, 48(1): 108-118.
doi: 10.1016/j.foodres.2012.03.006
[9] 李燕燕. 改性提高大米蛋白体外消化率的研究[D]. 无锡: 江南大学, 2015.
LI Y Y. Study on improving digestibility in vitro of rice protein by modification[D]. Wuxi: Jiangnan University, 2015. (in Chinese)
[10] WANG F, ZHANG Y Z, XU L, MA H L. An efficient ultrasound- assisted extraction method of pea protein and its effect on protein functional properties and biological activities. LWT-Food Science and Technology, 2020, 127: 109348.
doi: 10.1016/j.lwt.2020.109348
[11] O′SULLIVAN J, PARK M, BEEVERS J. The effect of ultrasound upon the physicochemical and emulsifying properties of wheat and soy protein isolates. Journal of Cereal Science, 2016, 69: 77-84.
doi: 10.1016/j.jcs.2016.02.013
[12] 李笑笑. 高场强超声波处理对大豆分离蛋白结构及乳化性的影响[D]. 广州: 华南理工大学, 2020.
LI X X. Effect of high intensity ultrasonic treatment on structure and emulsification of soy protein isolate[D]. Guangzhou: South China University of Technology, 2020. (in Chinese)
[13] CHANDRAPALA J, OLIVER C, KENTISH S, ASHOKKUMAR M. Ultrasonics in food processing. Ultrasonics Sonochemistry, 2012, 19(5): 975-983.
doi: 10.1016/j.ultsonch.2012.01.010
[14] 谢为峰, 罗丰收, 解超男, 王胜, 孔令明. 超声波处理对杏仁粕蛋白质理化和功能性质的改性研究. 农产品加工, 2016(9): 1-3, 7.
XIE W F, LUO F S, XIE C N, WANG S, KONG L M. Modification of ultrasonic treatment of almond meal protein physicochemical and functional properties. Farm Products Processing, 2016(9): 1-3, 7.(in Chinese)
[15] JIANG J, CHEN J, XIONG Y L. Structural and emulsifying properties of soy protein isolate subjected to acid and alkaline pH-shifting processes. Journal of Agricultural and Food Chemistry, 2009, 57(16): 7576-7583.
doi: 10.1021/jf901585n
[16] 丁芳芳, 田少君, 常慧敏, 章绍兵. 超声改性豌豆蛋白乳状液稳定性的研究. 中国油脂, 2020, 45(2): 76-110, 116.
DING F F, TIAN S J, CHANG H M, ZHANG S B. Modification of emulsion stability of pea protein by ultrasonic technology. China Oils and Fats, 2020, 45(2): 76-110, 116. (in Chinese)
[17] LERTITTIKUL W, BENJAKUL S, TANAKA M. Characteristics and antioxidative activity of Maillard reaction products from a porcine plasma protein-glucose model system as influenced by pH. Food Chemistry, 2007, 100(2): 669-677.
doi: 10.1016/j.foodchem.2005.09.085
[18] 曹云刚. 植物多酚对肉蛋白氧化稳定性和功能特性的影响机理及应用[D]. 无锡: 江南大学, 2016.
CAO Y G. Effects of plant-derived polyphenols on oxidative stability and functional properties of meat proteins: mechanism and application[D]. Wuxi: Jiangnan University, 2016. (in Chinese)
[19] LIU G, XIONG Y L. Electrophoretic pattern, thermal denaturation, and in vitro digestibility of oxidized myosin. Journal of Agricultural and Food Chemistry, 2000, 48(3): 624-630.
doi: 10.1021/jf990520h
[20] JIANG J, ZHANG X, TRUE A D, ZHOU L, XIONG Y L. Inhibition of lipid oxidation and rancidity in precooked pork patties by radical-scavenging licorice (Glycyrrhiza glabra) extract. Journal of Food Science, 2013, 78(11): C1686-C1694.
doi: 10.1111/1750-3841.12273
[21] WANG Q L, JIANG J, XIONG Y L. High pressure homogenization combined with pH shift treatment: a process to produce physically and oxidatively stable hemp milk. Food Research International, 2018, 106: 487-494.
doi: 10.1016/j.foodres.2018.01.021
[22] LI Y Y, LIU H T, LIU Q, KONG B H, DIAO X P. Effects of zein hydrolysates coupled with sage (Salvia officinalis) extract on the emulsifying and oxidative stability of myofibrillar protein prepared oil-in-water emulsions. Food Hydrocolloids, 2019, 87: 149-157.
doi: 10.1016/j.foodhyd.2018.07.052
[23] CARBONARO M, MASELLI P, NUCARA A. Relationship between digestibility and secondary structure of raw and thermally treated legume proteins: a Fourier transform infrared (FT-IR) spectroscopic study. Amino Acids, 2012, 43(2): 911-921.
doi: 10.1007/s00726-011-1151-4
[24] 夏轩泽, 李言, 钱海峰, 张晖, 齐希光, 王立. 豌豆蛋白乳化性及其改善研究进展. 食品与发酵工业, 2021, 47(2): 279-284.
XIA X Z, LI Y, QIAN H F, ZHANG H, QI X G, WANG L. Research progress on improvement the emulsification property of pea protein. Food and Fermentation Industries, 2021, 47(2): 279-284. (in Chinese)
[25] 包中宇. 超声波技术对大豆分离蛋白功能性质、结构及凝胶特性的影响[D]. 南昌: 南昌大学, 2015.
BAO Z Y. Effects of functional, structural and gelation property of soybean protein isolate treated by ultrasound[D]. Nanchang: Nanchang University, 2015. (in Chinese)
[26] ARZENI C, MARTINEZ K, ZEMA P, ARIAS A, PÉREZ O E, PILOSOF A M R. Comparative study of high intensity ultrasound effects on food proteins functionality. Journal of Food Engineering, 2012, 108(3): 463-472.
doi: 10.1016/j.jfoodeng.2011.08.018
[27] O'SULLIVAN J, MURRAY B, FLYNNC C, NORTONA I. The effect of ultrasound treatment on the structural, physical and emulsifying properties of animal and vegetable proteins. Food Hydrocolloids, 2016, 53: 141-154.
doi: 10.1016/j.foodhyd.2015.02.009
[28] BEN-HARB S, PANOUILLÉ M, HUC-MATHIS D, MOULIN G, SAINT-EVE A, IRLINGER F, BONNARME P, MICHON C, SOUCHON I. The rheological and microstructural properties of pea, milk, mixed pea/milk gels and gelled emulsions designed by thermal, acid, and enzyme treatments. Food Hydrocolloids, 2018, 77: 75-84.
doi: 10.1016/j.foodhyd.2017.09.022
[29] 卢菊慧. 豌豆蛋白的乳化特性及其影响机制研究[D]. 无锡: 江南大学, 2018.
LU J H. Study on the emulsifying property of pea protein and its influencing mechanism[D]. Wuxi: Jiangnan University, 2018. (in Chinese)
[30] GÜLSEREN I, GÜZEY D, BRUCE B D, WEISS J. Structural and functional changes in ultrasonicated bovine serum albumin solutions. Ultrasonics Sonochemistry, 2007, 14(2): 173-183.
doi: 10.1016/j.ultsonch.2005.07.006
[31] HU H, WU J H, EUNICE C Y, ZHU L, ZHANG F, XU X Y, FAN G, WANG L F, HUANG X J, PAN S Y. Effects of ultrasound on structural and physical properties of soy protein isolate (SPI) dispersions. Food Hydrocolloids, 2013, 30(2): 647-655.
doi: 10.1016/j.foodhyd.2012.08.001
[32] LIANG H N, TANG C H. Emulsifying and interfacial properties of vicilins: Role of conformational flexibility at quaternary and/or tertiary levels. Journal of Agricultural and Food Chemistry, 2013, 61(46): 11140-11150.
doi: 10.1021/jf403847k
[33] LIU G, XIONG Y L, BUTTERFIELD D A. Chemical, physical, and gel-forming properties of oxidized myofibrils and whey-and soy-protein isolates. Journal of Food Science, 2000, 65(5): 811-818.
doi: 10.1111/jfds.2000.65.issue-5
[34] CHEN N N, ZHAO Q Z, SUN W Z, ZHAO M M. Effects of malondialdehyde modification on the in vitro digestibility of soy protein isolate. Journal of Agricultural and Food Chemistry, 2013, 61(49): 12139-12145.
doi: 10.1021/jf404099y
[35] CHEN L, CHEN J S, REN J Y, ZHAO M M. Effects of ultrasound pretreatment on the enzymatic hydrolysis of soy protein isolates and on the emulsifying properties of hydrolysates. Journal of Agricultural and Food Chemistry, 2011, 59(6): 2600-2609.
doi: 10.1021/jf103771x
[36] 常慧敏, 杨敬东, 田少君. 超声辅助木瓜蛋白酶改性对米糠蛋白溶解性和乳化性的影响. 中国油脂, 2019, 44(4): 35-40.
CHANG H M, YANG J D, TIAN S J. Influence of ultrasound-assisted papain modification on solubility and emulsification of rice bran protein. China Oils and Fats, 2019, 44(4): 35-40.(in Chinese)
[37] 吴凡. 物理加工对牡蛎(Ostrea edulis)蛋白结构和功能性质的影响[D]. 大连: 大连工业大学, 2019.
WU F. Effects of physical processing on structure and functional properties of oysters protein[D]. Dalian: Dalian University of Technology, 2019. (in Chinese)
[38] TANG S Q, DU Q H, FU Z. Ultrasonic treatment on physicochemical properties of water-soluble protein from Moringa oleifera seed. Ultrasonics Sonochemistry, 2021, 71: 105357.
doi: 10.1016/j.ultsonch.2020.105357
[39] WANG Y S, ZHAO J, ZHANG W W, LIU C Q, JAUREGI P, HUANG M G. Modification of heat-induced whey protein gels by basic amino acids. Food Hydrocolloids, 2020, 100: 105397.
doi: 10.1016/j.foodhyd.2019.105397
[40] MAITY I, RASALE D B, DAS A K. Sonication induced peptide-appended bolaamphiphile hydrogels for in situ generation and catalytic activity of Pt nanoparticles. Soft Matter, 2012, 8(19): 5301-5308.
doi: 10.1039/c2sm25126d
[41] SUI X N, BI S, QI B K, WANG Z J, ZHANG M, LI Y, JIANG L Z. Impact of ultrasonic treatment on an emulsion system stabilized with soybean protein isolate and lecithin: Its emulsifying property and emulsion stability. Food Hydrocolloids, 2017, 63: 727-734.
doi: 10.1016/j.foodhyd.2016.10.024
[42] 王耀松, 张唯唯, 马天怡, 蔡敏, 张怡帆, 胡荣蓉, 唐长波. 丙二醛氧化对核桃分离蛋白结构及乳化性的影响. 中国农业科学, 2020, 53(16): 3372-3384.
WANG Y S, ZHANG W W, MA T Y, CAI M, ZHANG Y F, HU R R, TANG C B. Influence of oxidative modification by malondialdehyde on structure and emulsifying properties of walnut protein. Scientia Agricultura Sinica, 2020, 53(16): 3372-3384. (in Chinese)
[43] SHA L, KOOSIS A O, WANG Q L, TRUE A D, XIONG Y L. Interfacial dilatational and emulsifying properties of ultrasound- treated pea protein. Food Chemistry, 2021, 350: 129271.
doi: 10.1016/j.foodchem.2021.129271
[44] TAHA A, HU T, ZHANG Z, BAKRY A M, KHALIFA I, PAN S Y, HU H. Effect of different oils and ultrasound emulsification conditions on the physicochemical properties of emulsions stabilized by soy protein isolate. Ultrasonics Sonochemistry, 2018, 49: 283-293.
doi: 10.1016/j.ultsonch.2018.08.020
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