乳清浓缩蛋白对松仁蛋白溶解度及其乳液稳定性的影响
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(1.东北林业大学 生命科学学院, 黑龙江 哈尔滨 150040;2.黑龙江省林业科学研究所, 黑龙江 哈尔滨 150081;3.黑龙江省森林食品资源利用重点实验室, 黑龙江 哈尔滨 150040)

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Effect of Whey Protein Concentrate on Solubility and Emulsion Stability of Pine Kernel Protein
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(1. School of Life Science, Northeast Forestry University, Harbin 150040, China;2. Forestry Research Institute of Heilongjiang Province, Harbin 150081, China;3. Key Laboratory of Forest Food Resources Utilization of Heilongjiang Province, Harbin 150040, China)

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    摘要:

    为研究蛋白质-蛋白质相互作用对松仁蛋白(PKP)溶解度、结构和乳液性质的影响,以PKP和乳清浓缩蛋白(WPC)为研究对象,采用pH循环法制备PKP-WPC复合蛋白,利用SDS-PAGE、内源性荧光光谱、紫外-可见光谱、圆二色谱、荧光探针和ζ-电位等方法分析了复合蛋白结构特性和表面特性,再以PKP-WPC复合蛋白为原料,分别制备了油相体积分数为3%、10%和50%的乳液,并对制备的乳液性质进行了检测。结果表明:当WPC与PKP的质量比为1.0∶1.0,且体系pH值经历由7.0到12.0再回到7.0时的1次pH循环后,PKP的水溶性可从48.53%提高到92.43%。SDS-PAGE结果显示,PKP-WPC复合蛋白完整保留了PKP和WPC的亚基。内源性荧光光谱、紫外-可见光谱和圆二色谱结果表明,静电相互作用、疏水相互作用和氢键是驱动PKP和WPC相互作用的主要作用力,PKP与WPC相互作用使复合蛋白具有较高的结构韧性,抵抗酸诱导的构象折叠;WPC的加入改变了PKP的二级结构,α-螺旋、β-转角和无规卷曲结构的含量增加,而β-折叠结构相对含量降低。PKP-WPC复合蛋白具有较高的表面电荷(-34.74mV)来抵抗蛋白质的聚集。与由PKP制备的乳液相比,由PKP-WPC制备的乳液平均粒径和乳层析指数减小,ζ-电位绝对值增大,稳定性显著提高。乳液的性质因油相体积分数的不同而有较大的差异。油相体积分数为3%的复合乳液液滴小且分布均匀,稳定性好于油相体积分数为10%和50%的复合乳液。通过pH循环法,通过添加WPC,提高了PKP的溶解度,获得了稳定性较佳的PKP乳液,研究可为新型蛋白产品的研发提供理论基础,拓宽松仁蛋白在加工食品中的应用范围,推动PKP-WPC双蛋白乳液研究的发展。

    Abstract:

    To study effects of protein-protein interactions on solubility, structure and emulsion properties of pine kernel protein (PKP), PKP and whey protein concentrate (WPC) were used as research objects, and PKP-WPC complex protein was prepared by pH-cycle method. The structure and surface properties of complex protein were analyzed by SDS-PAGE, intrinsic fluorescence spectra, ultraviolet-visible spectra, circular dichroism, fluorescent probe and ζ- potential, and then emulsions with oil phase volume fractions of 3%, 10% and 50% were prepared using PKP-WPC complex protein as material, respectively. Finally, emulsion properties were tested. The results showed that when mass ratio of WPC addition to PKP was 1.0∶1.0 and pH of system went through a pH-cycle from 7.0 to 12.0 and back to 7.0, water solubility of PKP improved from 48.53% to 92.43%. SDS-PAGE results showed that PKP-WPC complex protein completely retained subunits of PKP and WPC. Intrinsic fluorescence spectra, ultraviolet-visible spectra and circular dichroism showed that electrostatic interaction, hydrophobic interaction and hydrogen bond were major forces driving the interaction between PKP and WPC, and interactions of PKP and WPC endowed complex protein with high structural toughness and resistance to acid-induced conformational folding. The addition of WPC changed secondary structure of PKP, the amount of α-helix, β-turn and random coil structures increased, while relative content of β-sheet structures decreased. PKP-WPC complex protein had higher surface charges (-34.74mV) to resist protein aggregation. Compared with emulsions prepared from PKP, emulsions prepared from PKP-WPC showed reduced mean particle diameter and creaming index, increased absolute value of ζ-potential, and significantly improved stability. Emulsions properties varied considerably depending on oil phase volume fraction. The composite emulsion with 3% oil phase volume fraction had small and uniformly distributed droplets, and its stability was better than composite emulsion with 10% and 50% oil phase volume fraction. The study improved PKP solubility by adding WPC through pH-cycle method, and obtained PKP emulsions with better stability. The study could provide a theoretical basis for the development of new protein products, broaden application of pine kernel protein in processed foods, and promote development of PKP-WPC double protein emulsion research.

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刘文超,杨凯,赵玉红.乳清浓缩蛋白对松仁蛋白溶解度及其乳液稳定性的影响[J].食品科学技术学报,2024,(1):57-68, 105.

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  • 收稿日期:2022-09-26
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  • 在线发布日期: 2024-02-06
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