Purification and properties of polyphosphate phosphohydrolase from chlorella ellipsoidea

Chlorellaellipsoidea로 부터 polyphosphate phosphohydrolase의 분리, 정제 및 성질

  • 임영복 (고려대학교 생물학과) ;
  • 이영록 (고려대학교 생물학과)
  • Published : 1983.09.01

Abstract

The presence of polyphosphate phosphohydrolase (PPPH) and tripolyphosphate phosphohydrolase (TPPH) in Chlorella ellipsoidea were confirmed from the cell-free extract of the algal cells and three forms of PPPH were isolated, purified, and measured Km-Vmax value and inhibitory effect by metal ions, respectively. PPPH was most active at pH7.2, whereas TPPH at pH 7.6. Both enzymes exhibited their maximum activity at $37^{\circ}C$. For the manifestation of catalytic activity, divalent, divalent metal ions are needed, and the best activator for both enzymes was $Co^{++}\;ions\;(10^{-3}M)$. These enzymes were inhibited by $Hg^{++}\;ions\;(10^{-3}M)$ considerably. PPPH from Chlorella ellipsoidea was purified by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-25, and gel filtration on Sephadex G-100, and some properties of the three different fraction with PPPH activity $(PPPH_1,\;PPPH_2,\;and\;PPPH_3)$ were found, i.e, PPPH has multiple form. The Km values of $PPPH_1,\;PPPH_2,\;and\;PPPH_3$ obstained were $6.25{\times}10^{-4}M,\;10^{-4}M-4/M,\;and\;3.33{times}10^{-4}M$ and Vmax were 3.33 mM/min, 3.33 mM/min, and 2.67 mM/min, respectively. It was shown that the types of inhibition of $Hg^{++} on the activities of three forms of PPPH were competitive inhibition.

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