Xanthine oxidase of Acinetobacter calcoaceticus RL2-M4: Production, purification and characterization.
Protein Expr Purif
; 160: 36-44, 2019 08.
Article
em En
| MEDLINE
| ID: mdl-30926462
ABSTRACT
Xanthine oxidase (EC 1.17.3.2) is a key enzyme of purine metabolism and has potential applications in food and pharmaceutical industries. In the present study, a new bacterial source of xanthine oxidase i.e. Acinetobacter calcoaceticus RL2-M4 with high oxidase activity was isolated from soil. The culture conditions were optimized with one variable at a time (OVAT) and response surface methodology (RSM) approaches included a minimal salt medium (MSM) of pH 7.0 supplemented with 0.8% yeast extract, 8.5â¯mM xanthine and incubation at 30⯰C for 24â¯h. Under these culture conditions 11.57 fold increase in the production of this enzyme was achieved. The enzyme was purified from A. calcoaceticus RL2-M4 using anion exchange chromatography to 8.18 fold with 31% yield and specific activity of 4.58 U/mg protein. SDS-PAGE analysis of the purified enzyme revealed that it was homodimer of 95â¯kDa and its native molecular mass was estimated to be 190â¯kDa. This enzyme was found to be stable at 35⯰C for 5â¯h. The purified xanthine oxidase of A. calcoaceticus RL2-M4 had Km 0.3â¯mM and Vmax 5.8 U/mg protein using xanthine as substrate. The activity and stability characteristic of xanthine oxidase of A. calcoaceticus RL2-M4 makes it a potentially good enzyme for industrial applications.
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MEDLINE
Assunto principal:
Proteínas de Bactérias
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Xantina Oxidase
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Acinetobacter calcoaceticus
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article