Production, purification and partial characterization of an endopolygalacturonase fromCryptococcus albidus var.albidus
- 1 July 1985
- journal article
- Published by Springer Nature in Antonie van Leeuwenhoek
- Vol. 51 (2) , 139-150
- https://doi.org/10.1007/bf02310007
Abstract
Cryptococcus albidus var.albidus produced an extracellular endo-polygalacturonase (poly (1,4-α-d-galacturonide) glycanohydrolase EC 3.2.1.15) when grown in a synthetic medium containing one of a variety of pectic substances or galacturonic acid. The highest level of enzyme activity (15.5 VU-ml−1) was obtained after 72 h of growth on 1.0% low-methoxyl pectin. The enzyme, purified by gel filtration (Sephadex G-100) after repeated ammonium sulphate precipitation and dialysis, showed only one band by polyacrylamide gel electrophoresis and had the following properties: mol wt (MWr) 41000 dal; isoelectric point (pl) = 8.10 ± 0.10; optimum temperature and pH for activity around 37°C and pH 3.75, respectively; pH stability in the pH range 4.0 to 8.0; complete heat inactivation after 10 min at 55°C; Km and Vmax values 5.7· 10−1 mg·ml−1 and 5.1 · 10−1 mmoles·min−1, respectively.Keywords
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