Determination of Rennin and Bovine Pepsins in Commercial Rennets and Abomasal Juices

A DEAE cellulose column chromatographic method is described to separate and to titrate rennin and bovine pepsin II from a mixture. Microgranular DEAE cellulose was equilibrated in .02m piper-azine buffer pH 5.3 and was placed on a 1×8cm column. The enzyme solution was poured on the column, as such,...

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Published inJournal of dairy science Vol. 55; no. 12; pp. 1641 - 1650
Main Authors Garnot, Pascaline, Thapon, J.L., Mathieu, C.M., Maubois, J.L., Dumas, B. Ribadeau
Format Journal Article
LanguageEnglish
Published Champaign Elsevier Inc 01.12.1972
Am Dairy Sci Assoc
American Dairy Science Association
Subjects
dairy products
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Summary:A DEAE cellulose column chromatographic method is described to separate and to titrate rennin and bovine pepsin II from a mixture. Microgranular DEAE cellulose was equilibrated in .02m piper-azine buffer pH 5.3 and was placed on a 1×8cm column. The enzyme solution was poured on the column, as such, or, if necessary, after dialysis against the same buffer. Elution of enzymes was by a linear gradient of NaCl (100ml of .15m NaCl in the first chamber and 100ml of .5m NaCl in the second chamber). The flow was about 80ml per hour. Rennin was eluted in the first fraction and pepsin II in the second fraction. This method was used to determine the enzymatic composition of several commercial rennet preparations and abomasal juices collected from fistulated calves.
ISSN:0022-0302
1525-3198
DOI:10.3168/jds.S0022-0302(72)85740-0