Purification and properties of cytidine deaminase from escherichia coli.
作者： G W Ashley , P A Bartlett
DOI: 10.1016/S0021-9258(18)90738-6
关键词:
摘要: A convenient and efficient procedure for the purification of cytidine deaminase (EC 3.5.4.5) from Escherichia coli is reported. The key step involves adsorption enzyme a crude ammonium sulfate fraction onto cytidine-containing affinity resin, followed by elution with 0.5 M borate buffer. Subsequent chromatography on DEAE-Sepharose results in an overall 1690-fold purification, yielding specific activity 118 units/mg. Cytidine has apparent molecular weight 54,000 as determined gel filtration, whereas sodium dodecyl sulfate-polyacrylamide electrophoresis shows band at 35,000. inhibited 5-(chloromercuri)cytidine kinetic behavior typical active-site-directed inactivation, KD = 0.09 mM kinact 1.25 min-1. protected against inactivation presence substrate, inhibition reversed high concentrations mercaptoethanol. This suggests that result mercaptide formation between mercury active-site thiol.
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