Susceptibility of phaseolin ( Phaseolus vulgaris) subunits to trypsinolysis and influence of dietary level of raw phaseolin on protein digestion in the small intestine of rats
Montoya, Carlos A. | Lallès, Jean-Paul | Beebe, Stephen | Souffrant, Wolfgang B. | Mölle, Daniel | Leterme, Pascal
The aim of the present work was (a) to investigate trypsinolysis of denatured purified T phaseolin (Phaseolus vulgaris) subunits by MS and (b) to test the effect of raw T phaseolin inclusion level in diets fed chronically to rats on digestion in the small intestine. The diets contained casein as the sole protein source, or casein substituted with 33, 67 and 100 % of purified T phaseolin. Rats were fed for 10 d and then euthanised. Digesta and tissues from the first and second halves of the small intestine were prepared for electrophoresis, immunoblotting and densitometry. α-Phaseolin subunit for the T phaseolin was more resistant to trypsinolysis than β-phaseolin subunit. Nearly intact phaseolin subunits (molecular weight, MW 44-54 kDa) and partially digested phaseolin fragments (MW 17-19 and 20-24 kDa) were identified in small intestinal digesta. The concentration of intact phaseolin and of most undigested phaseolin fragments in digesta increased in the second half of the small intestine with increasing phaseolin intake (P < 0·05-0·01). The concentration of phaseolin fragments of a MW of 21-22·5 and 23-24·5 kDa in the mucosa increased linearly (P = 0·016-0·084) when the level of the T phaseolin was increased in the diet. In conclusion, the present work provides evidence that denatured T phaseolin subunits display different trypsinolysis patterns in vitro. Moreover, a high intake of raw T phaseolin impacts digestion in the small intestine of rats.
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