The Comparison of the Characteristics of Partially Purified Internal Invertase by Mating Type in the Heterobasidiomycetous Yeast

이담자 효모균의 성접합형에 따른 세포내 Invertase의 성질 비교

  • Published : 1992.12.01

Abstract

This work was carried out to study physiological characteristics of Rhodosporidium toru[oides cells having two different mating types. The mating type A produces internal. cell wall-bound, and external invertases while type a produces only two invertases except external invertase. Comparing their characteristics after partial purification of internal invertases from both mating type cells, invertase from type a has decreased 15% of invertase activity only by $Mn^{2+}$ I while invertase from type A has been increased 11% of invertase activity by $Zn^{2+}$ and decreased 15% of invertase activity by $Mn^{2+}$ On the effect of enzyme inhibitor, invertase of type a was inhibited from 12% to 57% by 2-mercaptoethanol, sodium dodecyl sulfate, phenol. but invertase of type A was slightly inhibited only by phenol. The thermal stability of both invertases has showed steep inactivation at above $80^{\circ}C$ and their optimal temperatures were similar at $60^{\circ}C$ . Invertase from type A showed stability only on condition of acid from pH 3 to 6 and its opimal pH was 5.0, while invertase from type a showed stability at the wide range of pH 3-10 and its optimal pH was 4.0. And the $K_m$ values of invertases from type A and type a were $2.5{\times}10^3$M and$3.4{\times}10^3$M, respectively.

서로 다른 성접합형 A와 a를 가지는 Rhodosporidium toruloides가 생산하는 invertase를 조사한 결과, 접합형 A는 세포내, 세포벽 결합, 세포의 분비성 invertase의 3가지를 생산하였으나, 접합형 a는 세포외 invertase를 생산하지 않는 접합형 특이성이 있는 것을 알았다. 양 접합형 세포로부터 세포내 invertase를 부분정제하여 이들의 성질을 비교한 결과, 접합형 A세포 유래의 효소는 $Zn^{2+}$에 의하여 11의 활성 상승효과와Mn^{2+}$에 의한 42.2%의 감소를 보이는 반면, a세포 유래의 효소는 $Mn^{2+}$에 의한 15의 활성 감소만을 보였다. 효소 저해제의 효과에서 a세포의 invertase는 2-mercaptoethanol, sodium dodecyl sulfate, pheonl에만 약간 저해를 받을 뿐이었다. 이들 양 효소의 열안정성은 모두 $80^{\circ}C$ 이상에서 급격히 실활되는 양상을 보였으며 최적 온도가 $60^{\circ}C$로 둘다 비슷하였다. a세포 유래의 invertase는 pH 3에서 pH 10까지 넓은 pH의 범위에서 안정하며 최적 pH가 4.0인반면, A 세포의 invertase는 pH 3에서 pH 6까지 산성의 조건에서만 안정하였으며 최적 pH는 5.0으로 나타났다.그리고, A 세포와 atpvh 유래 invertase의 $K_m$값은 각각 $2.5{\times}10^3$M 과 $3.4{\times}10^3$M과 이였다.

Keywords

References

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