Purification and characterization of arginine decarboxylase from cucumber (Cucumis sativus) seedlings

Prasad, G. L. ; Adiga, P. R. (1985) Purification and characterization of arginine decarboxylase from cucumber (Cucumis sativus) seedlings Journal of Biosciences, 7 (3-4). pp. 331-343. ISSN 0250-5991

PDF - Publisher Version

Official URL: http://www.ias.ac.in/jarch/jbiosci/7/331-343.pdf

Related URL: http://dx.doi.org/10.1007/BF02716795


A simple, reproducible and rapid protocol for the purification of arginine decarboxylase fromCucumis sativus seedlings has been standardised. The purification steps involved ion-exchange chromatography on diethylaminoethyl-cellulose followed by gel filtration on Sephadex G-l 50. The purified enzyme preparation migrated as a single stainable band on Polyacrylamide gels at both basic and acidic pH, but under denaturing and reducing conditions on sodium dodecyl sulphate-polyacrylamide gels resolved into polypeptides of molecular weight 48,000,44,000 and 15,000. However, in the absence of 2-mercaptoethanol on electrophoresis on sodium dodecyl sulphate-polyacrylamide gels, the enzyme moved as single band with a molecular weight of 150,000. Evidence was obtained to indicate that these three polypeptides were probably derived from a single larger molecular weight enzyme. On storage of the purified protein, the 48,000 species was preferentially degraded to smaller polypeptides. The preliminary data suggested that the 48,000 and 44,000 species shared many common tryptic peptides as revealed by finger printing of the [125I ]-labelled protein. The purified enzyme was a glycoprotein and had a Km of 0.5 mM for arginine. Its activity was stimulated by dithiothrietol and pyridoxal phosphate. EDTA did not inhibit the enzyme activity. Mn2+ at 1 mM stimulated arginine decarboxylase activity but was inhibitory at higher concentration.

Item Type:Article
Source:Copyright of this article belongs to Indian Academy of Sciences.
Keywords:Arginine Decarboxylase; Peptide Mapping; Purification
ID Code:26765
Deposited On:08 Dec 2010 13:13
Last Modified:17 May 2016 10:04

Repository Staff Only: item control page