Abstract
Dye materials and cross linking agents were used for the determination of $\beta$-glucanase activities. The objective of this study was to prepare the blue coloured substrates which are sensitive, specific and simple for the determination of $\beta$-glucanase in malt and Bacillus subtilis K-4-3 enzymes. This method is based on the principle of measuring colorimetrically the split product of coloured and cross linked substrate. The best coupling of dye stuff of $\beta$-glucan was cibacron blue 3G-A and the colour released can suitably be measured at 623nm. Optimal concentration of dye and cross linking agents was 1.5g and 1.25$m\ell$ under 0.1N NaOH. The sensitivity comparison proved that the stained $\beta$-glucan method is much more sensitive than the DNS method to determine reducing sugar released by the enzyme.
$\beta$-Glucan 분해효소의 간편하며 예민한 활성도 측정방법으로 $\beta$-glucan에 색소와 cross linking agent를 접합시키는 변형기질 제조시 영향을 미치는 조건을 조사하고 맥아와 세균의 $\beta$-glucan분해효소 측정에 적응시켜 활성도를 측정한 결과는 다음과 같다. 1.0g $\beta$-glucan은 0.1N NaOH용액에서 색소 cibacron blue 3 G-A 1.5g과 cross linking agent인 1,4-butanedioldiglycidyl ether 1.25 $m\ell$을 90분간 끓여서 색소 접합기질로 제조하였을 때 $\beta$-glucanase 활성도 측정에 최적조건이었다. 변형기질은 pH5.3 에서 안정성을 보였으며 Bacillus subtilis K-4-3에서 추출한 효소액에 변형기질을 반응시켰을 때 간편하고 정확한 효소활성 측정이 가능하였다. 또한 색소방법을 DNS방법과 비교한 결과 색소방법이 $\beta$-glucan 분해효소 측정에 적당한 방법이었음이 입증되었다.