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二硫键和非共价键在热诱导乳清蛋白凝胶功能特性中的作用

The roles of disulphide and non-covalent bonding in the functional properties of heat-induced whey protein gels.

作者信息

Havea Palatasa, Carr Alistair J, Creamer Lawrence K

机构信息

Fonterra Research Centre, Private Bag 11029, Palmerston North, New Zealand.

出版信息

J Dairy Res. 2004 Aug;71(3):330-9. doi: 10.1017/s002202990400024x.

Abstract

Heat-induced gelation (80 degrees C, 30 min or 85 degrees C, 60 min) of whey protein concentrate (WPC) solutions was studied using transmission electron microscopy (TEM), dynamic rheology and polyacrylamide gel electrophoresis (PAGE). The WPC solutions (150 g/kg, pH 6.9) were prepared by dispersing WPC powder in water (control), 10 g/kg sodium dodecyl sulphate (SDS) solution or 10 mM-dithiothreitol (DTT) solution. The WPC gels containing SDS were more translucent than the control gels, which were slightly more translucent than the gels containing DTT. TEM analyses showed that the SDS-gels had finer aggregate structure (approximately equal to 10 nm) than the control gels (approximately equal to 100 nm), whereas the DTT-gels had a more particulate structure (approximately equal to 200 to 300 nm). Dynamic rheology measurements showed that the control WPC gels had storage modulus (G) values (approximately equal to 13,500 Pa) that were approximately equal to 25 times higher than those of the SDS-gels (approximately equal to 550 Pa) and less than half those of the DTT-gels after cooling. Compression tests showed that the DTT-gels were more rigid and more brittle than the control gels, whereas the SDS-gels were softer and more rubbery than either the control gels or the DTT-gels. PAGE analyses of WPC gel samples revealed that the control WPC solutions heated at 85 degrees C for 10 min contained both disulphide bonds and non-covalent linkages. In both the SDS-solutions and the DTT-solutions, the denatured whey protein molecules were in the form of monomers or small aggregates. It is likely that, on more extended heating, more disulphide linkages were formed in the SDS-gels whereas more hydrophobic aggregates were formed in the DTT-gels. These results demonstrate that the properties of heat-induced WPC gels are strongly influenced by non-covalent bonding. Intermolecular disulphide bonds appeared to give the rubbery nature of heat-induced WPC gels whereas non-covalent bonds their rigidity and brittle texture.

摘要

采用透射电子显微镜(TEM)、动态流变学和聚丙烯酰胺凝胶电泳(PAGE)对乳清蛋白浓缩物(WPC)溶液的热诱导凝胶化(80℃,30分钟或85℃,60分钟)进行了研究。通过将WPC粉末分散在水(对照)、10g/kg十二烷基硫酸钠(SDS)溶液或10mM二硫苏糖醇(DTT)溶液中来制备WPC溶液(150g/kg,pH6.9)。含有SDS的WPC凝胶比对照凝胶更半透明,对照凝胶比含有DTT的凝胶稍半透明。TEM分析表明,SDS凝胶的聚集结构比对照凝胶更精细(约10nm),而对照凝胶约为100nm,而DTT凝胶具有更颗粒状的结构(约200至300nm)。动态流变学测量表明,冷却后,对照WPC凝胶的储能模量(G)值(约13500Pa)约为SDS凝胶(约550Pa)的25倍,不到DTT凝胶的一半。压缩试验表明,DTT凝胶比对照凝胶更坚硬、更脆,而SDS凝胶比对照凝胶或DTT凝胶更软、更具弹性。对WPC凝胶样品的PAGE分析表明,在85℃加热10分钟的对照WPC溶液中同时含有二硫键和非共价键。在SDS溶液和DTT溶液中,变性的乳清蛋白分子均为单体或小聚集体形式。很可能在更长时间的加热过程中,SDS凝胶中形成了更多的二硫键,而DTT凝胶中形成了更多的疏水聚集体。这些结果表明,热诱导WPC凝胶的性质受到非共价键的强烈影响。分子间二硫键似乎赋予了热诱导WPC凝胶橡胶状的性质,而非共价键赋予了它们刚性和脆性质地。

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