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在不同截留分子量的透析袋中评估β-乳球蛋白/多糖在体外胃和胰水解过程中的相互作用。

Beta-lactoglobulin/polysaccharide interactions during in vitro gastric and pancreatic hydrolysis assessed in dialysis bags of different molecular weight cut-offs.

作者信息

Mouécoucou J, Villaume C, Sanchez C, Méjean L

机构信息

Laboratoire des Sciences Animales, ENSAIA-INPL, USC INRA 2, Avenue de la Forêt de Haye BP 172, F-54505 Vandoeuvre-lès-Nancy Cedex 5, France.

出版信息

Biochim Biophys Acta. 2004 Jan 22;1670(2):105-12.

Abstract

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.

摘要

研究了阿拉伯胶、低甲基化(LM)果胶或木聚糖在0和50 wt.%水平对β-乳球蛋白(β-lg)消化率的影响,以及体外水解过程中两种大分子之间的相互作用。蛋白水解在一个涉及两步水解的系统中进行:要么单独使用胃蛋白酶,要么先使用胃蛋白酶,然后在截留分子量(MWCO)为1000或8000 Da的透析袋中使用胰蛋白酶/胰凝乳蛋白酶(T/C)混合物。通过氮释放以及水解后两个透析袋截留物的SDS-PAGE电泳来评估消化率。比浊法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几乎对胃蛋白酶消化具有抗性,尽管胃蛋白酶活性降低,但多糖增加了氮释放。这是由于多糖与β-lg之间形成了静电复合物,减少了β-lg的聚集,增加了其溶解度。使用MWCO为1000 Da的透析袋测定时,多糖显著降低了β-lg的T/C消化率,且未改变其酶活性。使用MWCO为8000 Da的透析袋时,未检测到多糖对β-lg消化率有显著影响。电泳图谱未显示与所用透析袋相关的截留物谱有差异。这表明在水解的第二步中,多糖与小于8000 Da的氨基酸或肽之间可能发生非特异性相互作用。

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