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LOCALIZATION OF DISACCHARIDASES IN EXTRACTS OF NEUROSPORA AFTER ELECTROPHORESIS IN POLYACRYLAMIDE GELS

F. I. EILERS 1, J. ALLEN 1, E. P. HILL 1, and A. S. SUSSMAN 1

1 Departments of Botany and Zoology, University of Michigan, Ann Arbor, Michigan

A technique for localizing the activity of disaccharidases on acrylamide gels is described. The technique is a modification of that of Dahlqvist and Brun and employs as the chromogenic reagent a mixture of glucose oxidase, nitro BT and phenazine methosulfate, with a disaccharide as the substrate. A formazan is deposited at the site of enzyme activity, thereby permitting localization. Through this means the invertase activity of several strains of Neurospora crassa has been investigated. Two of the "standard" strains used showed two invertases whereas a variant derived from one of these had only one. A trehalase which runs ahead of the invertases also has been demonstrated. A band is formed with cellobiose but is at the position of one of the invertases so that this enzyme may be of mixed specificity. No band was formed with lactose as substrate.

Submitted on October 31, 1963


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