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Original Article

Identification of yeast phase of pathogenic fungi by the specificity of their aminopeptidase(s)

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Pages 132-141 | Published online: 09 Jul 2009
 

Abstract

Specificity of aminopeptidase(s) was fluorimetrically determined in the yeast phase of Histoplasma capsulatum, H. duboisii, H. farciminosum, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Candida albicans and Cryptococcus neoformans. After individually incubating each of 26 amino acid-β-naphthylamides with each yeast, the amount of each amino acid-β-naphthylamide hydrolyzed was determined by measuring the free naphthylamine. This resulted in a reproducible profile of the aminopeptidase(s) for each fungus when medium, growth time, size of inoculum and incubation period were standardized. This technique provided a rapid and specific means of identification and differentiation among these yeasts. Specific amino acids were identified from the profile of each yeast as those rapidly liberated by the aminopeptidase(s) of that yeast. These amino acids delineated the amino acid requirement for the normal growth of each yeast.

Zusammenfassung

Die Spezifität der Aminopeptidasen wurde fluorometrisch bei der Hefephase von Histoplasma capsulatum’ H. duboissii, H. farciminosum, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Candida albicans und Cryptococcus neoformans bestimmt.

Nachdem jede dieser Hefen auf jede einzelne von 26 Aminosäure-β-naphthylamidverbindungen eingewirkt hatte, bestimmten wir jeweils die Menge des hydrolisierten Aminosäure-β-naphthylamides, ndem wir das freie Naphthylamin ma βen. Das führte zu einem reproduzierbaren “Leistungsprofil” der Aminopeptidase (n) jedes Pilzes, wenn Nährboden, Wachstumszeit, Inokulatmenge und Bebrütungsdauer standardisiert waren. Diese Technick lieferte eine schnelle und spezofische Möglichkeit der Identifizierung und Differenzierung der verschiedenen Hefen. Durch das “Leistungsprofil” jeder Hefeart indentifizierten wir spezifische Aminosäuren, und zwar jene, die durch die Aminopeptidase jeder einzelnen Hefe rasch freigesetzt wurden. Diese Aminosäuren kennzeichnen das Aminosäurebedürfnis für das normale Wachstum jeder Hefeart.

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