The activities of some metabolic enzymes in the intestines of germ-free and conventional chicks.

Abstract

The metabolic enzymes alkaline phosphatase (EC 3.1.3.1), acid phosphatase (EC 3.1.3.2) and isocitrate dehydrogenase (EC 1.1.1.42) were measured in mucosal homogenates and these enzymes, together with glucose-6-phosphatase (EC 3.1.3.9), were measured in homogenates of isolated enterocytes from germ-free (GF) and conventional (CV) chicks which were either fed continuously until they were killed or were subjected to a 16 h fast before killing. The intestine of the GF chicks was generally lighter than that of the CV controls. The activity of alkaline phosphatase was greater in the mucosal homogenates of the CV chicks compared with the GF birds, but the concentrations of acid phosphatase and isocitric dehydrogenase were not different in the two groups. In the isolated enterocytes the concentration of all enzymes expressed per mg DNA, except alkaline phosphatase, was higher in the GF chicks. Expressed per mg protein there was no significant difference in enzyme activity in the two groups. Fasting caused a reduction in intestinal weight and total mucosal protein in both groups but the reduction was greater in the GF chicks compared with the CV controls. In the GF chicks, fasting caused a significant fall in acid phosphatase and isocitric dehydrogenase activities of the mucosal homogenate, whereas in the CV chicks only acid phosphatase fell to a significant extent. In the isolated enterocytes feeding caused a marked fall in protein per mg DNA in the CV chicks; fasting tended to reduce enzyme concentrations in the GF chicks but to have less effect in the CV group except for alkaline phosphatase where there was a marked rise in activity. It is suggested that the difference in enzyme activities in the mucosal homogenates and isolated enterocytes might resulted from (a) the presence of a much greater lamina propria in the CV compared with the GF chicks and (b) the greater mitotic activity in the fed CV chicks yielding a much larger number of smaller immature cells.