Application of high-performance chromatographic and electrophoretic methods to the purification and characterization of glucose oxidase and catalase from penicillium chrysogenum

Kjell ove Eriksson, Iordanka Kourteva, Kunquan Yao, Jia li Liao, Frence Kilár, Stellan Hjertén, Grigori Chaga

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21 Citations (Scopus)

Abstract

The high resolving power of the preparative and analytical high-performance chromatographic and electrophoretic methods resently developed in this laboratory for the separation of biopolymers has been demonstrated by the purification and characterization of glucose oxidase and catalase from Penicillium chrysogenum. Crude glucose oxidase was purified to homogeneity in one step by high-performance hydrophobic-interaction chromatography )HIC) on a pentylagarose column. Crude catalase was purified by a combination of HIC and high-performance anion-exchange chromatography on 3-diethylamino-2-hydroxypropylagarose. The homogeneity of the enzymes was monitored by high-performance electrophoresis and free zone electrophoresis. the p values of these two enzymes determined by isoelectric focusing in the high-performance electrophoresis apparatus were 4.2 and 6.5, respectively. Their molecular weights were determined by high-performance molecular sieve chromatography on an agarose column. Glucose oxidase has a molecular weight of 175 000 and probably consists of two identical subunits, as sodium dodecyl sulphate polyacrylamide gel electrophoresis gave a molecular weight of around 72 000. The molecular weight of catalase, which is probably composed of non-dentical subunits, as indicated by sodium dodecyl sulphate electrophoresis, is around 320 000. Some other characteristics of these two enzymes were also investigated, e.g., electrophoretic mobility, pH stability and optimum pH.

Original languageEnglish
Pages (from-to)239-249
Number of pages11
JournalJournal of Chromatography A
Volume397
Issue numberC
DOIs
Publication statusPublished - Jun 26 1987

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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