Purification and properties of phenylalanyl aminopeptidase synthesised by Pseudomonas sp.

Abstract:

:Intracellular aminopeptidase synthesized by a soil strain of Pseudomonas sp. was purified 323-fold using the following procedure: saturation with ammonium sulfate, separation by preparative electrophoresis, anion-exchange chromatography and gel filtration chromatography. Molecular weight of the enzyme determined according to the latter method was 57 kDa. Aminopeptidase showed a high substrate specificity and affinity to Phe-beta-naphtylamide (Phe-beta-NA) as a substrate. A considerable inhibition of the enzymatic activity by iodoacetamide and p-chloromercuribenzoate (p-CMB) led to the conclusion that it was a cysteine aminopeptidase. Hydrosulphide compounds markedly stabilised the enzyme. Ethylenediaminetetra-acetic acid (EDTA), a metalloenzyme inhibitor, caused a double increase in the phenylalanyl aminopeptidase activity.( )Mg(2+) ions activated the enzyme to a negligible extent, whereas Co(2+), Cu(2+), Cd(2+) and Pb(2+) ions contributed to its inhibition. The highest enzymatic activity was observed at 37 degrees C and pH 7.0.

journal_name

J Basic Microbiol

authors

Jankiewicz U,Bielawski W

doi

10.1002/1521-4028(200208)42:4<260::AID-JOBM260>3.0

subject

Has Abstract

pub_date

2002-01-01 00:00:00

pages

260-7

issue

4

eissn

0233-111X

issn

1521-4028

journal_volume

42

pub_type

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