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Journal of microbiology and biotechnology v.15 no.3, 2005년, pp.652 - 655   피인용횟수: 2

Cloning and Expression of Isocitrate Lyase, a Key Enzyme of the Glyoxylate Cycle, of Candida albicans for Development of Antifungal Drugs

SHIN DONG-SUN    (Natural Products Research Institute, College of Pharmacy, Seoul National University   ); KIM SANGHEE    (Natural Products Research Institute, College of Pharmacy, Seoul National University   ); YANG HYEONG-CHEOL    (Department of Dental Biomaterials Science, College of Dentistry, Seoul National University   ); OH KI-BONG    (Natural Products Research Institute, College of Pharmacy, Seoul National University, Graduate School of Agricultural Biotechnology, Seoul National University, Center for Agricultural Biomaterials, Seoul National University  );
  • 초록

    This paper describes the development of an enzymatic assay system for the identification of inhibitors of isocitrate lyase (ICL), one of the key enzymes of the glyoxylate cycle that is considered as a new target for antifungal drugs. A 1.6 kb DNA fragment encoding the isocitrate lyase from Candida albicans ATCC10231 was amplified by PCR, cloned into a vector providing His-Patch-thioredoxin-tag at the N-terminus, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The molecular mass of the purified ICL was approximately 62 kDa, as determined by SDS-PAGE, and the enzyme activity was directly proportional to incubation time and enzyme concentration. The effects of itaconate-related compounds on ICL activity were also investigated. Among them, itaconic acid, 3-nitropropionate, and oxalate had strong inhibitory activities with $IC_{50}$ values of 5.8, 5.4 and $8.6\;{mu}g/ml$ , respectively. These inhibitors also exhibited antifungal activity on YPD agar media containing acetate as a sole carbon source, albeit at high concentration. The results indicate that the C. albicans ICL may be a regulatory enzyme playing a crucial role in fungal growth and is a prime target for antifungal agents.

  • 주제어

    Candida albicans .   glyoxylate cycle .   isocitrate lyase .   gene expression .   in vitro assay .   inhibitors.  

  • 참고문헌 (20)

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  • 이 논문을 인용한 문헌 (2)

    1. 2006. "" Journal of microbiology and biotechnology, 16(10): 1597~1604     
    2. 2007. "" Archives of pharmacal research : a publication of the Pharmaceutical Society of Korea, 30(8): 955~961     

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