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The journal of microbiology v.35 no.2, 1997년, pp.109 - 116  

Purification and Characterization of Extracellular Aspartic Proteinase of Candida albicans

Na, Byoung-Kuk    (Department of Biology, Faculty of Natural Science, Chung-Ang University   ); Lee, Seong-Il    (Department of Biology, Faculty of Natural Science, Chung-Ang University   ); Kim, Sin-Ok    (Korean Institute of Tuberculosis   ); Park, Young-Kil    (Korean Institute of Tuberculosis   ); Bai, Gill-Han    (Korean Institute of Tuberculosis   ); Kim, Sang-Jae    (Korean Institute of Tuberculosis   ); Song, Chul-Yong    (Department of Biology, Faculty of Natural Science, Chung-Ang University  );
  • 초록

    An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45 $^{\circ}C$ . The addition of divalent cations, $Ca^{2+}$ , Zn $^{2+}$ and $Mg^{2+}$ , resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe $^{2+}$ , Ag $^{2+}$ and Cu $^{2+}$ . With BSA as substrate, an apparent $K_m$ was determined to be 7 $\times$ 10 $^{-7}$ M and $K_i$ , using pepstatin A as an inhibitor, was 8.05 $\times$ 10 $^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P $_1$ position, but the enzyme activity was highly reduced when the P $_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.


  • 주제어

    Candida albicans .   Aspartic proteinase .   Purification .   Characterization.  

  • 참고문헌 (34)

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