在不同分子量切断的透析袋中评估体外胃和胰腺水解过程中β -乳球蛋白/多糖的相互作用。

Biochimica et biophysica acta Pub Date : 2004-01-22
J Mouécoucou, C Villaume, C Sanchez, L Méjean
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引用次数: 0

摘要

研究了阿拉伯树胶、低甲基化(LM)果胶或木聚糖在0和50% wt.%水平下对β -乳球蛋白(β -lg)消化率的影响,以及这两种大分子在体外水解过程中的相互作用。蛋白水解是在一个涉及两步水解的系统中进行的:要么单独胃蛋白酶,要么胃蛋白酶之后是胰蛋白酶/凝乳胰蛋白酶(T/C)混合物,在透析袋中,分子量切断(MWCO)为1000或8000 Da。通过水解后两个透析袋的N释放和保留物的SDS-PAGE电泳来估计消化率。浊度测量监测了两步水解过程中混合物的结构演变。结果表明,β -lg几乎抵抗消化,多糖增加了N的释放,尽管降低了胃蛋白酶的活性。这是由于多糖和β -lg之间形成静电配合物,这减少了β -lg的聚集,增加了其溶解度。使用MWCO为1000 Da的透析袋测定,多糖显著降低了β -lg T/C的消化率,而不改变其酶活性。采用mwco8000da透析袋检测,多糖对β -lg消化率无显著影响。电泳模式没有显示保留物的轮廓与使用的透析袋有关的差异。这表明在多糖与小于8000 Da的氨基酸或肽水解的第二步可能发生非特异性相互作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Beta-lactoglobulin/polysaccharide interactions during in vitro gastric and pancreatic hydrolysis assessed in dialysis bags of different molecular weight cut-offs.

The effects of gum arabic, low methylated (LM) pectin or xylan at levels of 0 and 50 wt.% on beta-lactoglobulin (beta-lg) digestibility were studied as well as the interactions between the two macromolecules during in vitro hydrolysis. The proteolysis was performed in a system involving a two-step hydrolysis: either pepsin alone, or pepsin followed by a trypsin/chymotrypsin (T/C) mixture in dialysis bags with molecular weight cut-offs (MWCO) 1000 or 8000 Da. Digestibility was estimated by the N release and by a SDS-PAGE electrophoresis of retentates from the two dialysis bags after hydrolysis. Turbidimetric measurements monitored the structural evolution of mixtures during the two-step hydrolysis. Results showed that beta-lg was almost resistant to peptic digestion and that polysaccharides increased the N release despite a reduction of pepsin activity. This is due to the formation of electrostatic complexes between polysaccharides and beta-lg, which reduced beta-lg aggregation, increasing its solubility. The polysaccharides reduced significantly the beta-lg T/C digestibility as determined using a dialysis bag with a MWCO 1000 Da, without a modification of their enzymatic activities. No significant effect of polysaccharides on the beta-lg digestibility was detected using the dialysis bag with a MWCO 8000 Da. The electrophoresis pattern did not show differences in the profile of retentates in relation with the dialysis bag used. This suggests that non-specific interactions could occur during the second step of hydrolysis between polysaccharides and amino acids or peptides smaller than 8000 Da.

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