Heat inactivation and sephadex chromatography of the small-intestine disaccharidases of the chick.
1. The maltase, sucrase, isomaltase and palatinase activities of the chick small intestine are localized in particles that sediment when centrifuged at 100000g for 90min. 2. Solubilization of the particle-bound disaccharidases without loss of activity was achieved by digestion with papain. Trypsin was less effective and caused a preferential solubilization of the sucrase, isomaltase and palatinase activities. 3. On Sephadex G-200 columns, the solubilized preparations yielded two disaccharidase peaks. The first peak was eluted close to the void volume of the column and contained all the sucrase, isomaltase and palatinase activities and some of the maltase activity. The remainder of the maltase activity was eluted beyond the total volume of the column. 4. Precipitation with ethanol did not affect the behaviour of the disaccharidases of gel filtration. 5. The maltase activity of the second peak on rechromatography in a buffer containing 0.01m-maltose was eluted close to the void volume. 6. Similar pH optima but different K(m) values were obtained for the maltase activities of the two peaks. 7. Heat-inactivation studies showed that the first peak contained two disaccharidase enzymes; one hydrolysed sucrose and maltose and the other hydrolysed isomaltose, palatinose and maltose. The second peak contained three disaccharidase enzymes all specific for the hydrolysis of maltose. 8. It is proposed that the intestinal disaccharidases of the chick exist in the form of two complexes: a sucrase-isomaltase complex and a maltase complex.
D000818:Animals; D011232:Chemical Precipitation; D002645:Chickens; D002845:Chromatography; D002850:Chromatography, Gel; D000431:Ethanol; D006026:Glycoside Hydrolases / Q000032:analysis*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007421:Intestine, Small / Q000201:enzymology*; D007700:Kinetics; D008320:Maltose / Q000378:metabolism; D010206:Papain / Q000378:metabolism; D013393:Sucrase / Q000032:analysis; D014357:Trypsin / Q000378:metabolism
D000431:Ethanol; D008320:Maltose; D006026:Glycoside Hydrolases; D013393:Sucrase; D014357:Trypsin; D010206:Papain