Characterization of a phenol oxidase from Cryptococcus neoformans var. neoformans.
In Cryptococcus neoformans, enzymic oxidation of various catechols leads to melanin, a proposed virulence factor. A phenol oxidase enzyme of Cryptococcus neoformans var. neoformans produced at 25 C has been purified from an ultracentrifugal supernatant of an extract of broken cells. Hydrophobic interaction chromatography followed by anion-exchange column chromatography allowed purification of the phenol oxidase. The molecular weight of the enzyme estimated by gel filtration was about 80,000 and a dimeric species (Mw = 160,000) was suggested. The isoelectric point of the protein was approximately 4.1. An NH2-terminal 31 amino acid sequence was determined using phenol oxidase electroblotted onto a PVDF membrane after nondenaturing gel electrophoresis. Upon searching the Peptide Institute (Osaka) data base, no proteins with high degrees of homology were found.
Ikeda
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R
Department of Microbiology, Meiji College of Pharmacy, Tokyo, Japan.
Shinoda
T
T
Morita
T
T
Jacobson
E S
ES
D000595:Amino Acid Sequence; D002847:Chromatography, Agarose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003455:Cryptococcus neoformans / Q000201:enzymology*; D007526:Isoelectric Point; D008543:Melanins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014442:Monophenol Monooxygenase / Q000302:isolation & purification*
D008543:Melanins; D014442:Monophenol Monooxygenase