Isoenzymes of chloroperoxidase from Caldariomyces fumago

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Abstract
Chloroperoxidase (EC 1.11.1.10) was purified from Caldaromyces fumago strains ATCC 16373 and CM1 89362 grown in defined and complex media. Analysis of the enzyme by polyacrylamide gel electrophoresis at pH 3, sodium dodecyl sulfate – polyacrylamide gel electrophoresis, and isoelectric focussing in polyacrylamide gels revealed two forms of the enzyme, the major form representing greater than 80% of the total protein. When the fungi were grown in synthetic media, more of the enzyme was found in the major band than when the fungi were grown in the presence of malt extract. Enzyme activity was demonstrated both by staining gels in situ with 3,3′,5,5′-tetramethylbenzidine and by spectrophometric assay of material eluted from gel slices. There were no obvious differences between the enzyme from C. fumago ATCC 16373 and from C. fumago CMI 89362. Commercial samples, included for comparative purposes, contained three or four enzymically active species which separated on polyacrylamide gel electrophoresis at pH 3 and electrofocussing but migrated as a close double band on sodium dodecyl sulfate – polyacrylamide gel electrophoresis. It is believed that prolonged storage in the unfrozen state is responsible for the conversion to multiple forms.