PURIFICATION AND PHYSICOCHEMICAL PROPERTIES OF β‐GALACTOSIDASE FROM Kluyveromyces fragilis
Authors:
- 1 March 1978
- journal article
- research article
- Published by Wiley in Journal of Food Science
- Vol. 43 (2) , 584-591
Abstract
β‐Galactosidase was purified to apparent homogeneity from cell‐free extracts of Kluyveromyces fragilis by acetone precipitation, pseudo‐affinity chromatography, hydroxylapatite chromatography and DEAE‐Sephadex A‐50 chromatography. The molecular weight was 201,000 and the partial specific volume was 0.715. No carbohydrate was detected. Electron microscopy indicated a molecular diameter of about 160Å with 9‐10 subunits present. Isoelectric point of the major isozyme was 5.1. The extinction coefficient was 1.58 cm 3 mg protein ‐1 at 280 nm. The amino‐acid composition was quite different from the β‐galactosidase of Escherichia coli. Enzyme activity was rapidly lost on incubation with p ‐chloromercuribenzoate; formation of 5‐6 moles of mercaptide per mole of protein gave complete loss of activity.Keywords
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