Blood Lysate Staining, a New Microscopic Method for Diagnosis of Fungemia Using Peripheral Blood

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  • YAMAMOTO Tetsuro
    Alcoholic Beverages Research Laboratories, Takara Shuzo Co., Ltd. Mie Institute for Microbiology, Nichinichi Food Co., Ltd.
  • NOHARA Kumiko
    Research Center for Medical Mycology, Teikyo University School of Medicine
  • UCHIDA Katsuhisa
    Research Center for Medical Mycology, Teikyo University School of Medicine
  • YAMAGUCHI Hideyo
    Research Center for Medical Mycology, Teikyo University School of Medicine Department of Microbiology, Teikyo University School of Medicine

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  • Purification and Characterization of Secretory Proteinase of <i>Candida albicans</i>

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Proteolytic activity of medically important yeasts was tested in both YCB-BSA agar and medium. All of 134 strains of Candida albicans, 13 of 18 strains of Candida tropicalis and 11 of 18 strains of Candida parapsilosis had this activity, while none of 52 Candida glabrata strains or of 11 Cryptococcus neoformans strains tested had proteolytic activity. Strains of C. albicans fell into five groups based on the level and time-course of in vitro proteinase productivity. Five strains randomly selected from each group were tested for pathogenicity in mice. The strain possessing the strongest pathogenicity was used to purify proteinase. The molecular weight of the proteinase was approximately 44, 000 daltons and its isoelectric point was pH 4.2. Optimal pH of the proteinase was 3.2 and the enzyme was stable below pH 7.0 and lost its activity above pH 8.0 at 37C in a 60-min incubation. The 23 amino acid sequence of the proteinase N-terminus was determined.

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