본 연구는 rotifer, B. rotundiformis를 대상으로 소화효소 실험을 하기 위해 이들이 가지고 있는 소화효소의 최고 활성 조건을 확인하기 위해 수행하였다. rotifer, B. rotundiformis의 $alpha$-amylase, total alkaline Protease, trypsin 및 TG-lipase는 Tris-HCl buffer 보다 phosphate-NaOH buffer 안정적인 효소활성을 보였다. $alpha$-amylase, trypsin 및 TG-lipase는 pH 8.0에서, total alkaline proteaset pH 7.0에서 높은 효소 활성을 나타내었다. $alpha$-amylase 활성은 $40^{circ}C$에서 가장 높은 활성을 보였으며, total alkaline pretense와 trypsin은 $55{~}60^{circ}C$의 온도에서 높은 활성을 나타내었다. 반면 TG-lipase 활성은 $25{~}30^{circ}C$의 낮은 온도에서 활성이 높았다. $alpha$-amylase, total alkaline pretense, trypsin 및 TG-lipase의 활성의 적정 기질 농도는 $3.5\%$ starch, $ .6%$ azo-casein, $87.5{mu}M$ BApNA and 81.2 mM olive oil이었다. $alpha$-amylase, total alkaline protease, trypsin 및 TG-lipase의 활성의 적정 반응시간은 40, 60, 30 and 25 min으로 나타났다. 본 연구 결과에서 얻어진 자료는 rotifer, B. rotundiformis의 소화효소 연구를 위한 기초 자료로 이용될 것이다.
This study was investigated the condition of their maximum activity to assay the enzymes of rotifer, Brachionus rotundiformis의 $alpha$-amylase, total alkaline Protease, trypsin and TG-lipase activities of rotifer were higher and more sensitive in phosphate-NaOH buffer than Tris-HCl buffer. $alpha$-amylase, trypsin and TG-lipase activities were appeared the maximum at pH 8.0, and total alkaline protease activity showed the maximum activity at pH 7.0. $alpha$-amylase activity showed the highest activity at $40^{circ}C$, and total alkaline protease and trypsin activities were assayed the highest at $55{~}60^{circ}C$. However, TG-lipase activity was appeared the highest at $25{~}30^{circ}C$. The optimum substrate concentration of enzyme activity of a-amylase, total alkaline protease, rypsin and TG-lipase were $3.5\%$ starch, $ .6%$ azo-casein, $87.5{mu}M$ BApNA and 81.2 mM olive oil, respectively. The optimum reaction time of enzyme activity of $alpha$-amylase, total alkaline protease, trypsin and TG-lipase were increased up to 40, 60, 30 and 25 min., respectively. The data obtained in this study could be used for the digestive enzyme research of rotifer, B. rotundiformis.
