L-asparaginase of Thermus thermophilus: purification, properties and identification of essential amino acids for its catalytic activity.

Abstract:

:L-asparaginase EC 3.5.1.1 was purified to homogeneity from Thermus thermophilus. The apparent molecular mass of L-asparaginase by SDS-PAGE was found to be 33 kDa, whereas by its mobility on Sephacryl S-300 superfine column was around 200 kDa, indicating that the enzyme at the native stage acts as hexamer. The purified enzyme showed a single band on acrylamide gel electrophoresis with pI = 6.0. The optimum pH was 9.2 and the Km for L-asparagine was 2.8 mM. It is a thermostable enzyme and it follows linear kinetics even at 77 degrees C. Chemical modification experiments implied the existence ofhistidyl, arginyl and a carboxylic residues located at or near active site while serine and mainly cysteine seems to be necessary for active form.

journal_name

Mol Cell Biochem

authors

Pritsa AA,Kyriakidis DA

doi

10.1023/a:1011066129771

keywords:

subject

Has Abstract

pub_date

2001-01-01 00:00:00

pages

93-101

issue

1-2

eissn

0300-8177

issn

1573-4919

journal_volume

216

pub_type

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