초록
근해 연안 토양으로부터 Chitinase 활성이 우수한 균주를 분리하여 Aeromonas salmonicida로 동정하였으며, 분리균주의 효소생산 최적조건은 colloidal chitin 1.26 , tryptone 2.95 , $MgSO_4-7H_20$0.15 , $K_2HP0_4$, 0.15 , pH8.5, 27'C에서 48시간 진탕배양하였을 때였다. 효소의 정제는 배양액으로부터 ammonium sulfate 침전, affinity adsorption, hydroxylapatite chromatography, gel filtration을 통해 수율 29.7 , 정제도 18.5배의 정제효소를 얻었다. 정제된 chitinase의 최적온도와 pH는 $50^{\circ}C$와 7.0이었고 pH 안정성은 pH5.0-9.0 사이였고 $50^{\circ}C$까지 안정하였으며 Km값은 1.276mg/ml, 분자량은 200,000 daltons으로 확인되었다.
A chitinase-producing bacterium, Aeromonas salmonicida YA7-625, was isolated from domestic seashore muds. The preferable medium composition for the production of chitinase was as follows: colloidal chitin 1.26% (w/v), tryptone 2.95% (w/v), $MgSO_4-7H_20$ 0.15% (w/v) and $K_2HP0_4$, 0.15% (w/v) (pH 8.5). The highest enzyme production was observed after cultivation of 48 hours at 27OC. The chitinase of Aeromonas salmonicida YA7-625 was purified successively by ammonium sulfate precipitation, affinity adsorption, hydroxylapatite column chromatography and gel filtration. The optimal temperature and pH for the activity of purified chitinase were $50^{\circ}C$ and 7.0, respectively. The molecular weight of purified chitinase was ca. 200,000 daltons and apparent Km value of it was 1.276 mglml on colloidal chitin.