Immobilization of human intestinal mucosa enzymes on sepharose

✍ Scribed by P. Würsch

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 485 KB
• Volume: 77
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90312-8

✦ Synopsis

Digestive enzymes from human intestinal mucosa were solubilized by Triton X-100 and papain and covalently bound to cyanogen bromide-activated Sepharose 4-B gel. Triton X-100 solubilized most of the activities, and 39.1 to 63.5% were immobilized on the carrier. The other enzymes, still bound on the microvilli, were subsequently solubilized by papain but then the yield of immobilization reached only 11.0 to 17.6%. The enzyme-Sepharose gel was freeze-dried with a filler and stored without loss of activity. The rate of hydrolysis of di-and hisaccharides, dipeptides, andp-nitrophenylphosphate was measured by incubation on a small column containing less than 0.03 U of immobilized activities. The enzymatic multiplicity and catalytic properties of the intestinal mucosa enzymes were fully recovered on the carrier. This method is proposed for routine evaluation of the digestibility of dipeptides and synthetic disaccharides. 265