Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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A Study on the Purification and Characteristics of Branched-Chain Amino Acid Aminotransferase in Cultural Mycelia of Cordyceps militaris

번데기동충하초 균사 중의 Branched-Chain Amino Acid Aminotransferase의 분리정제 및 그 특성에 관한 연구

  • Kim, Sung-Tae (Department of Laboratory Medicine, Dongguk Univercity Ilsan Bulkyo Hospital) ;
  • Park, Chung-Oh (Department of Biomedical Laboratory Science, Seoul Health College)
  • 김성태 (동국대학교 일산불교병원 진단검사의학과) ;
  • 박정오 (서울보건대학 임상병리과)
  • Published : 2005.08.31
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Abstract

The optimum conditions of Cordyceps militaris mycelial growth, purification and characteristics of branched-chain amino acid aminotransferase [BCAT(EC 2.6.1.42)] in this mycelium were studied. Optimum pH, temperature and medium of culture of mycelia were 5.5, $22.5^{\circ}C$ and Hamada medium (HM), respectively. BCAT in homogenate of this mycelia was precipitated by 20-40% saturated solution of ammonium sulfate and then purified by DEAE (diethylaminoethyl)-Sephadex A-50 column chromatography with linear concentration gradient and Sephadex G-200 gel filtration. A single band of purified enzyme was detected on SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis). Optimum pH and temperature of BCAT were found to be 7.8 and $29^{\circ}C$, respectively. It showed activity toward L-leucine, L-isoleucine and L-valine as a substrate. The Km values of this enzyme for L-leucine were determined to be 5.88 mM for L-leucine.

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