Disaccharidase activities in camel small intestine: biochemical investigations of maltase-glucoamylase activity.
Disaccharidases (maltase, cellobiase, lactase, and sucrase), alpha-amylase, and glucoamylase in the camel small intestine were investigated to integrate the enzymatic digestion profile in camel. High activities were detected for maltase and glucoamylase, followed by moderate levels of sucrase and alpha-amylase. Very low activity levels were detected for lactase and cellobiase. Camel intestinal maltase-glucoamylase (MG) was purified by DEAE-Sepharose and Sephacryl S-200 columns. The molecular weight of camel small intestinal MG4 and MG6 were estimated to be 140,000 and 180,000 using Sephacryl S-200. These values were confirmed by SDS-PAGE, where the two enzymes migrated as single subunits. This study encompassed characterization of MGs from camel intestine. The Km values of MG4 and MG6 were estimated to be 13.3 mM and 20 mM maltose, respectively. Substrate specificity for MG4 and MG6 indicated that the two enzymes are maltase-glucoamylases because they catalysed the hydrolysis of maltose and starch with alpha-1,4 and alpha-1,6 glycosidic bonds, but not sucrose with alpha-1,2 glycosidic bond which was hydrolyzed by sucrase-isomaltase. Camel intestinal MG4 and MG6 had the same optimum pH at 7.0 and temperature optimum at 50 degrees C and 40 degrees C, respectively. The two enzymes were stable up to 50 degrees C and 40 degrees C, followed by strong decrease in activity at 60 degrees C and 50 degrees C, respectively. The effect of divalent cations on the activity of camel intestinal MG4 and MG6 was studied. All the examined divalent cations Ca(2+), Mn(2+), Mg(2+), Co(2+) and Fe(3+) had slight effects on the two enzymes except Hg(2+) which had a strong inhibitory effect. The effect of different inhibitors on MG4 and MG6 indicated that the two enzymes had a cysteine residue.
Mohamed
Saleh A
SA
Molecular Biology Department, National Research Centre, Cairo, Egypt. saleh38@hotmail.com
Fahmy
Afaf S
AS
Salah
Hala A
HA
D000818:Animals; D002162:Camelus / Q000378:metabolism*; D002413:Cations, Divalent / Q000378:metabolism / Q000494:pharmacology; D004186:Disaccharidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D065089:Glycoside Hydrolase Inhibitors; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007421:Intestine, Small / Q000737:chemistry / Q000201:enzymology*; D008670:Metals / Q000378:metabolism / Q000494:pharmacology; D013379:Substrate Specificity; D000520:alpha-Glucosidases / Q000302:isolation & purification / Q000378:metabolism*
D002413:Cations, Divalent; D004791:Enzyme Inhibitors; D065089:Glycoside Hydrolase Inhibitors; D008670:Metals; D004186:Disaccharidases; D000520:alpha-Glucosidases