An in vitro digestion method to assess the quality of proteins based on enzymatic hydrolysis is presented. The method consists of a two-step proteolysis at 37 degrees, a 30-minute incubation of the protein with pepsin at pH 1.9, followed by pancreatin digestion at pH 8 for 24 hours in a dialysis bag with a 1000 molecular weight cutoff for the continuous elimination of the digested products into a replaceable buffer. Three types of buffer replacement were tried. These were, in increasing order of efficiency: intermittent with either six changes (infrequent buffer replacement) or eleven changes (frequent buffer replacement) or continuously circulated at the rate of 212 ml/hour (continuous buffer replacement). By using three protein sources, i.e., casein, soybean and rapeseed proteins, it was found that the degree of digestion (dialyzed N) as well as the regularity of the process were markedly improved by increasing the frequency of buffer replacement. In spite of different pepsin digestibilities, as determined from the production of trichloroacetic acid-soluble N, the digestion of the three proteins gave equivalent results when the best procedure of buffer replacement was used (continuous buffer replacement). The deleterious effect of heat and alkali treatment on protein digestion was readily shown by this procedure and indicated casein to be a very heat-sensitive protein as compared to the others.
An in vitro digestion method was proposed for studying protein digestion. Protein (casein) was submitted to peptic proteolysis in a closed system followed by hydrolysis with pancreatic enzymes in a "digestion cell" with continuous elimination of.digested products by dialysis. The peptic digestion was performed with a pepsin source of high specific activity (3152 units/mg protein) with an enzyme:substrate (E:S) ratio of 1:250. The second-step of proteolysis was carried out with pancreatin for 6 hr at an E:S ratio of 1:25. A 10 mM sodium phosphate buffer, pH 7.5, was used as the circulating dialysis buffer.
The effects of purified and semipurified dietary fiber supplements on glycemia and insulinemia were measured simultaneously with their effects on digestive tract function in the rat. An insoluble fiber (cellulose) and four soluble fibers (guar gum, carboxymethylcellulose, mustard mucilage, and oat beta-glucan) were added separately to a fiber-free solid diet and fed to Sprague-Dawley rats for 10 days. Guar gum and oat beta-glucan reduced the food intake, whereas cellulose increased it. Guar gum reduced weight gain. Cellulose increased the protein efficiency ratio. After a 13-h fast, glycemia and insulinemia were measured 45, 90, 210, and 360 min after the beginning of a voluntary short meal. Addition of fibers did not change the glycemic response, but soluble fibers significantly decreased insulinemia 45 min after the meal. All fibers significantly delayed gastric emptying, cellulose and mustard mucilage being the most effective. Dry matter contents of the small intestine were increased especially by guar gum and oat beta-glucan. All fibers seemed to slow down small intestinal transit and decreased intestinal absorption. In the present experimental situation, both gastric and intestinal components played a role in the hypoinsulinic effect of dietary fibers. The intestinal component appeared to be more determinant for all soluble fibers, except mustard mucilage where the gastric component was more important.
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