Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Substrate Variety of a Non-metal Dependent Tagatose-6-phosphate Isomerase from Staphylococcus aureus

Staphylococcus aureus 유래 비금속성 이성화효소인 Tagatose-6-phosphate Isomerase의 기질다양성

  • Oh Deok-Kun (Department of Bioscience and Biotechnology, Sejong University) ;
  • Ji Eun-Soo (Major in Biotechnology, The Catholic University of Korea) ;
  • Kwon Young-Deok (Major in Biotechnology, The Catholic University of Korea) ;
  • Kim Hye-Jung (Department of Bioscience and Biotechnology, Sejong University) ;
  • Kim Pil (Major in Biotechnology, The Catholic University of Korea)
  • 오덕근 (세종대학교 생명공학과) ;
  • 지은수 (가톨릭대학교 생명공학전공) ;
  • 권영덕 (가톨릭대학교 생명공학전공) ;
  • 김혜정 (세종대학교 생명공학과) ;
  • 김필 (가톨릭대학교 생명공학전공)
  • Published : 2005.06.01
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Abstract

To investigate the substrate variety of a putative non-metal dependent isomerase, the tagatose-6-phosphate isomerase (E.C. 5.3.1.26) structural genes (lacB; 510bp and lacA; 430bp) of Staphylococcus aureus were subcloned and co-expressed. Based on the substrate configuration, various aldoses were surveyed for substrate of ketose isomerization. Among the 10 aldoses tested, D-ribose and D-allose were isomerized by the enzyme. The subunit A and B showed more than $95\%$ activity for D-ribose and $75\%$ for D-allose in the presence of 1mM EDTA compared with non-EDTA conditions, which implying tagatose-6-phosphate isomerase is a non-metal dependent isomerase. Each of subunit A or subunit B alone showed no activity for any of the substrates tested. The affinity constant ($K_m$) of tagatose-6-phosphate isomerase against D-ribose and D-allose were 26 mM and 142 mM, respectively.

비금속성 이성화효소로 추정되는 Staphylococcus aureus의 tagatose-6-phosphate isomerase(E.C. 5.3.1.26)의 기질다 양성을 조사하기 위해서 그 구조유전자(lacB;510bp와 lacA;430bp)를 대장균에서 동시발현하였다. 알려진 기질 이외에 D-ribose와 D-allose에 대해 이성화활성이 새롭게 관찰되었다. EDTA 1 mM 존재하에서도 D-ribose와 D-allose에 대하여 각각 EDTA 비존재 조건에 대비하여 $95\%,\;75\%$의 이성화활성을 나타내는 것으로 미루어 tagatose-6-phosphate isomerase가 비금속성 이성화효소임을 밝혔다. 이때 lacA 또는 lacB의 단독발현시에는 이성화활성이 전혀 밝견되지 않았다. D-Ribose와 D-allose에 대한 기질친화상수 ($K_m$)은 각각 26 mM와 142 mM였다.

Keywords

References

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