Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Characterization and Effect of Metal Ion on Activity of Phytase from Rat intestinal Mucosa

흰쥐 소장 점막 phytase의 특성 및 활성에 미치는 금속 이온의 영향

  • 양원진 (동아대학교 유전공학연구소) ;
  • 손흥대 (동아대학교 생명자원과학대학 농생물학과)
  • Published : 1997.06.01
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Abstract

Phytase(myo-inositol hexkisphosphate phosphohydrolase ; EC 3.1.3.8) was purified from the mucoas of rat intestinal. The molecular weight of enzyme was determined to be 160kDa by sephacryl S-200 gel filtration. Analysis of the purified enzyme o SDS-polyacrylamide gel electrophoresis(SDS-PAGE) showen that it was composed of two different subunits and the molecular weight of its subunit was found to be 70kDa and 90kDa respectively, indicating that this enzyme is hetrodimer. The enzyme activities were activated in the presence of $ MgCl_{2}$, but inhibited by $ZnCl_{2}$, $MnCl_{2}$, and EDTA. The substrates tested, phytase showed the highest affinity for the enzyme at the physiological ph. The Km value for phytic acid(inositol-hexakisphosphate)was 0.31 mM at pH 7.4. rat intestinal mucosa phytase seems to play an important in the metabolism of inositol.

Phytase(myo-inositol hexakisphosphate phosphohydro-lase;EC 3.1.3.8)는 흰주 소장 점막으로 부터 분리${\cdot}$정제 하였다. 이 정제된 효소를 Sephacryl S-200 gel filtration방법으로 측정한 분자량으니 160kDa이고, 순도 및 이 효소의 서브유니트를 SDS-polycrylamide gel전기영동법(SDS-PAGE)으로 조사한 결과 서브유니트 구조는 분자량이 10kDa와 90kDa으로 구성된 hetrodimer(이종이량체)임을 알 수 있었다. 그리고 $ MgCl_{2}$ 존재 하에서는 효소 활성이 증가하나 $ZnCl_{2}$, $MnCl_{2}$, 및 EDTA존재 하에서는 효소 활성이 저해되었다. 기질 트이성과 pH범위에서 기질인 phytic acid(inositol-hexakispho-sphate)에 대해 높은 친화력을 보였다. Phytic acid에 대한 Km값은 pH 7.4에서 0.31mM이다. 따라서 흰쥐의 소장 점막 phytase는 주로 inositol의 대사계에서 중요한 역할을 하는 것으로 생각된다.

Keywords

References

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