Isolation of psychrotrophic microorganism producing soymilk-clotting enzyme from marine fish

생선으로부터 분리한 두유 응고 효소 생산 호냉성 미생물

A psychrotrophic microorganism isolated from Alaska pollack (Theragra chalcoramma) produced soymilk-clotting enzyme(s) with relatively low proteolytic activity. The isolate No. 268 was tentatively identified as Pseudomonas sp. Soymilk-clotting activity of the crude enzyme solution was observed at temperatures ranging from 20 to $60^{\circ}C$ and the optimum temperature was $40^{\circ}C$. When the crude enzyme solution was preincubated for 30 minutes, the clotting activity was stable at temperatures up to $30^{\circ}C$ and 75% of the activity was retained at $40^{\circ}C$. The clotting activity was decreased as the pH of soymilk was increased from 5.8 to 7.3.

생선 시료로부터 대두유를 배지원으로 $10^{\circ}C$에서 증식배양하고, 조효소액의 효소활성을 조사하여 대두유 응고활성이 높고 단백질 분해활성이 낮은 호냉성 세균을 최종 선발하여 Pseudomonas sp.로 잠정 동정하였다. 조효소액의 대두유 응고활성은 $20{\sim}60^{\circ}C$에서 관찰되었고 최적온도는 $40^{\circ}C$였으며, 단백질 분해활성의 최적온도는 $40^{\circ}C$였고 $30^{\circ}C$이하 및 $60^{\circ}C$ 이상에서는 단백질 분해활성이 나타나지 않았다. 조효소의 활성은 pH$5.8{\sim}7.3$ 범위에서 pH가 증가함에 따라 감소되었다.