外加旋转流场对蛋白分子空间构象的影响
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(浙江工商大学食品与生物工程学院 杭州 310018)

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国家自然科学基金项目(31171713,31701650);浙江省自然科学基金项目(LY22C200009,LY17C200004)


Effect of An Applied Rotational Flow Field on the Spatial Conformation of Protein Molecules
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(The School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018)

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    摘要:

    目的:研究不同外加旋转流场对溶菌酶分子空间结构的影响,有利于高蛋白质含量的食品加工。方法:借助旋转流变仪在溶菌酶溶液中施加剪切流场,采用圆二色光谱、紫外吸收及二阶导数光谱、荧光发射光谱以及动态光散射技术进行表征,研究剪切流场对溶菌酶分子二级结构、三级结构的影响。结果:在剪切流场转速高于200 s-1或持续作用时间大于1.5 h时,溶菌酶分子α-螺旋结构比例降低以及无序结构比例增高,紫外吸收光谱发生蓝移,原本存在于分子内部的色氨酸残基极性增强。这种变化是不可逆的且随着剪切流场的转速增大、作用时间的延长和溶菌酶分子的浓度降低表现得更为显著。结论:剪切流场会对溶菌酶分子造成不可逆的构象改变,且剪切速率越大,作用时间越长,构象改变越显著。

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    Objective: This paper mainly studies the influence of different applied rotating flow fields on the spatial structure of lysozyme, which can be used to guide the food processing of high protein content food. Methods: A rotational rheometer was used to apply a shear flow field in lysozyme solution. Circular dichroism spectroscopy, ultraviolet absorption and second derivative spectroscopy, fluorescence emission spectroscopy and dynamic light scattering were used to characterize the effects of shear flow field on the secondary structure and tertiary structure of lysozyme molecules. Results: When the shear flow speed was higher than 200 s-1 or the shear flow duration was longer than 1.5 h, the proportion of α-helix structure decreased and the proportion of disorder structure increased, the UV absorption spectrum changed blue shift, and the polarity of tryptophan residue in the molecular was enhanced. This change was irreversible and became more significant with the increase of shear speed, the extension of action time and the decrease of lysozyme concentration. Conclusion: The shear flow field can cause an impossible inverse conformational change of lysozyme, and the larger the shear rate and the longer the interaction time, the more significant the conformational change.

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潘伟春,顾斐娜,赵萌迪,蔡枫烨,温娅,徐海星,牛付阁.外加旋转流场对蛋白分子空间构象的影响[J].中国食品学报,2024,24(12):48-56

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  • 收稿日期:2023-12-26
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  • 在线发布日期: 2025-01-23
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