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Purification and characterization of an extracellular adenosine deaminase from Nocardioides sp. J‐326TK
Author(s) -
HK Jun,
TS Kim,
Yeeh Y.
Publication year - 1994
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1994.tb00317.x
Subject(s) - chemistry , sephadex , adenosine deaminase , enzyme , adenosine , chromatography , extracellular , size exclusion chromatography , stereochemistry , biochemistry
An extracellular adenosine deaminase was isolated from the culture supernatant of Nocardioides sp. J‐326TK and purified 193‐fold to homogeneity. It had a specific activity of 4677 units/mg at 37 degrees C, was a monomeric protein as judged by SDS/PAGE, and was characterized with respect to M(r) (80,000 and 72,000 by gel filtration on Sephadex G‐200 and SDS/PAGE respectively), pH optimum (6.0), temperature optimum (50 degrees C) and pI (7.6). The adsorption spectrum of the enzyme had a maximum at 280 nm and a minimum at 250 nm. The enzyme was stable at pH 6.5‐7.5 and at temperatures below 30 degrees C. Adenosine and 2'‐deoxyadenosine were deaminated and the respective Km values were 0.22 and 0.20 mM, but the enzyme was not active on adenine and 6‐(gamma gamma'‐dimethylallylamino)purine riboside. The enzyme reaction was promoted by Fe3+ and Sn2+, but potently inhibited by Hg2+, Ag2+, o‐phenanthroline and pentachlorophenol, and noticeably inhibited by 8‐bromoadenosine, theobromine and theophylline.