CHROMATOGRAPHIC BEHAVIOR OF GLUCOSE-1-OXIDASE AND GLUCOSE-2-OXIDASE FROM FUNGAL STRAINS ON IMMOBILIZED METAL-CHELATES

Citation
V. Pacheco et A. Karmali, CHROMATOGRAPHIC BEHAVIOR OF GLUCOSE-1-OXIDASE AND GLUCOSE-2-OXIDASE FROM FUNGAL STRAINS ON IMMOBILIZED METAL-CHELATES, Journal of industrial microbiology & biotechnology, 21(1-2), 1998, pp. 57-64
Citations number
34
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biothechnology & Applied Migrobiology
Volume
21
Issue
1-2
Year of publication
1998
Pages
57 - 64
Database
ISI
SICI code
Abstract
Glucose 2-oxidase (EC 1.1.3.10) from Coriolus versicolor and Phaneroch aete chrysosporium and glucose 1-oxidase (EC 1.1.3.4) from Aspergillus niger bound to a CU(II)-IDA column in the pH range of 6-8. However, g lucose 1-oxidase from Penicillium amagasakiense bound only partially t o a CU(II)-IDA column at pH 8.0. Metal chelates containing either Ni(I I) or Zn(II) were useful in the adsorption of glucose P-oxidase from P hanerochaete chrysosporium. The binding of glucose 2-oxidase from P. c hrysosporium to Ni(II) and Zn(II)-IDA agarose columns increases as a f unction of pH of the buffer system. The adsorption of glucose oxidases on metal(II)-IDA chelates was due to the available histidine residues on enzyme molecules since the addition of imidazole in the buffer sys tem abolished the binding of glucose oxidases to these columns. Both g lucose oxidases from C.versicolor, P. chrysosporium and A, niger were purified in one step by immobilized metal affinity chromatography on m etal(II)-IDA agarose columns with a recovery of enzyme activity in the range of 80-91%. Purified preparations of glucose oxidases from funga l strains were apparently homogeneous on native PAGE and SDS-PAGE. Imm obilized metal affinity chromatography was used to separate glucose 1- oxidase from the 2-oxidase on metal(II)-IDA agarose columns which was confirmed by analysis of the reaction products by HPLC. The different chromatographic behaviour of glucose oxidases on metal(II)-IDA chelate s is apparently due to the number and spatial distribution of availabl e histidine residues on these enzyme molecules.