한국 재래 간장에서 분리한 Scopulariopsis brevicaulis가 생성하는 ${\alpha}-galactosidase$의 특성 및 작용양상

Characteristics and Action Pattern of ${\alpha}-galactosidase$ from Scopulariopsis brevicaulis in Korean Traditional Meju

한국 재래 간장으로부터 분리한 Scopulariopsis brevicaulis가 생성하는 ${\alpha}-galactosidase$의 최적 생산조건은 tryptone 1.5%, $NH_4NO_3$ 0.2%, raffinose 2.5%, $KH_2PO_4$ 0.5%, yeast extract 0.5%, pH 7.0, $27^{\circ}C$에서 3일간 배양 했을 때였다. ${\alpha}-galactosidase$의 최적 pH는 7.0, 최적온도는 $27^{\circ}C$였으며, $pH\;6.0{\sim}8.0$범위와 $40^{\circ}C$이하에서 안정하였다. $Ag^{2+},\;Hg^{2+},\;Cu^{2+}$ 등의 금속이온 및 p-chloromercuribenzoic acid, Iodine에 의해 강하게 저해되어 본 효소의 catalytic부위에 -SH기가 있음이 추정되었다. Km값은 1.9 mM, Vmax값은 $9.66{\times}10^2\;{\mu}M/min$이었다. 배양기간에 따른 raffinose의 분해양상을 high performance liquid chromatography로 살펴본 결과 배양초기에 raffinose가 분해되어 sucrose, glucose, fructose가 관찰되었다. 배양기간이 경과함에 따라 raffinose가 감소하였고 sucrose, glucose, fructose도 소실됨을 확인하였다.

The optimum culture condition of Scopulariopsis brevicaulis for the production of ${\alpha}$-galactosidase was as follows: Tryptone 1.5%, $NH_4NO_3$ 0.2%, Raffinose 2.5%, $KH_2PO_4$ 0.5%, yeast extract 0.5%, pH 7.0, $27^{\circ}C$. The optimum pH and temperature for the enzyme activity of ${\alpha}$-galactosidase producing Scopulariopsis brevicaulis were pH 7.0 and $27^{\circ}C$, respectively. The enzyme was relatively stable at $pH\;6.0{\sim}8.0$ and at temperature below $40^{\circ}C$. The activity of the enzyme was inhibited by $Ag^{2+},\;Hg^{2+},\;Cu^{2+}$, p-chloromercuribenzoic acid and Iodine. These results would indicate the presence of -SH groups in the catalytic site of the enzyme. Km value was 1.9 mM for $p-nitrophenyl-{\alpha}-D-galactopyranoside$ and Vmax value was $9.66{\times}10^2\;{\mu}M/min$. Sugar constituents of culture broth were identified by HPLC that the enzyme liberated sucrose, glucose and fructose from raffinose and raffinose was significantly decreased.