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从荧光葡聚糖探针的扩散推断出玉米淀粉颗粒内部结构的异质性。

Heterogeneity in maize starch granule internal architecture deduced from diffusion of fluorescent dextran probes.

机构信息

Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, ARC Centre of Excellence in Plant Cell Walls, The University of Queensland, St. Lucia, Brisbane, Queensland 4072, Australia.

出版信息

Carbohydr Polym. 2013 Apr 2;93(2):365-73. doi: 10.1016/j.carbpol.2012.12.017. Epub 2013 Jan 7.

Abstract

Heterogeneity in maize starch granules was investigated by studying the diffusion of fluorescent dextran probes (20, 70 and 150kDa) inside granules using fluorescence recovery after photobleaching combined with confocal microscopy. Access of probes to the interior of granules was greatly enhanced by limited (2.4%) amylolysis. The diffusion of probes within granules was found to be either 'fast' with diffusion coefficients in the order of 10(-6)cm(2)s(-1) or 'slow' with diffusion coefficients in the order of 10(-7)cm(2)s(-1), independent of the size of dextran probes or prior treatment of the granules by α-amylase. Results were compared with observations of pores and channels in granules by electron microscopy and by confocal microscopy after labelling with 8-amino-1,3,6-pyrenetrisulfonic acid. It is proposed that there is an inherent heterogeneity of internal architecture in maize starch granules due to the presence or absence in individual granules of (a) pores leading to a central cavity, resulting in 'fast' diffusion of dextran probes and (b) accessibility of the starch polymer matrix to dextran probes, leading to 'slow' diffusion behaviour. The observed heterogeneity of maize starch granule porosity has implications for chemical modification reactions and the kinetics of digestion with amylases.

摘要

使用荧光恢复后光漂白结合共聚焦显微镜研究了荧光葡聚糖探针(20、70 和 150 kDa)在颗粒内部的扩散,以研究玉米淀粉颗粒的异质性。通过有限的(2.4%)淀粉水解极大地促进了探针进入颗粒内部。发现探针在颗粒内的扩散要么是“快速”的,扩散系数在 10(-6)cm(2)s(-1)左右,要么是“缓慢”的,扩散系数在 10(-7)cm(2)s(-1)左右,与葡聚糖探针的大小或α-淀粉酶对颗粒的预处理无关。结果与电子显微镜观察到的颗粒中的孔和通道以及用 8-氨基-1,3,6-三嗪磺酸标记后通过共聚焦显微镜观察到的孔和通道进行了比较。提出由于个体颗粒中存在或不存在(a)导致中心腔的孔,导致葡聚糖探针的“快速”扩散,以及(b)葡聚糖探针对淀粉聚合物基质的可及性,导致“缓慢”扩散行为,因此玉米淀粉颗粒内部结构存在固有异质性。观察到的玉米淀粉颗粒孔隙率的异质性对化学修饰反应和与淀粉酶的消化动力学具有重要意义。

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