Characterization of Human ${\beta}-Carotene$ 15,15-dioxygenase Isolated from Recombinant Escherichia coli

유전자 재조합 기술에 의하여 제조된 인간 ${\beta}-carotene$ 15,15'-dioxygenase의 반응특성

  • Published : 2004.06.30

Abstract

Characteristics of human ${\beta}-carotene$ 15,15'-dioxygenase isolated by recombinant DNA technology was elucidated. Optimal pH and temperature were 9.0 and $40^{\circ}C$, respectively. Enzyme activity was temperature-sensitive. Enzyme was stable at pH 6.0-9.0 for 24 hr and under $5^{\circ}C$. Half-life of enzyme at $35^{\circ}C$ was 40 min. Crude preparations of enzyme were inhibited by ferrous ion-chelating agent and sulfhydryl-binding agent. GSH offsets inhibitory effect of PCMB. With increasing substrate concentrations, enzyme activity gave typical Michaelis-Menten curve, Based on Hanes-Woolf plot of data, $K_{m}\;and\;V_{max$ were $3.39{\times}10^{6}\;M\;and\;1.2\;pmol/mg$ protein/min, respectively.

Keywords

${\beta}-carotene$ 15,15'-dioxygenase;recombinant DNA technology;optimal pH and temperature;$K_m$;$V_{max}$

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