길항세균 C. violaceum strain C-61과 strain C-72, S. marcescens, A. hydrophila 및 A. caviae의 배양여액을 ammonium sulfate으로 침전시켜 chitin 분해효소의 생산 및 특성을 조사하였다. C. violaceum strain들은 28℃에서 3일, S. marcescens는 6일, A. hydrophila 및 A. caviae는 2일 배양하였을 경우에 가장 많은 chitinase를 생산하였다. 최적 배양 조건에서, C. violaceum strain C-61의 chitinase 활성은 strain C-72 보다 약 1.5배, S. marcescens 보다 약 5.5배, A. hydrophila 및 A. caviae 보다 각각 12.0배 및 11.3배 더 높았다. 그러나, N,N’-diacetylchitobiase 활성은 C. violaceum strain C-61 보다 S. marcescens에서 3.2배 더 높았고, A. hydrophila 및 A. caviae에서는 매우 낮았다. glycol chitin이 함유된 gel에서, C. violaceum strain들의 chitinase는 54-, 52-, 50-, 37-kDa의 4 isoform이 검출되었는데, S. marcescens는 58-, 52-, 48-, 38-kDa의 4 isoform, A. hydrophila는 70-, 58-, 54-kDa의 3 isoform, 그리고 A. caviae는 90-, 79-, 71-, 63-, 58-, 38-kDa의 6 isoform이 검출되었다. C. violaceum strain C-61의 chitinase는 pH 7.0과 50℃에서 가장 활성이 높았고, pH 5.0~10.0에 2시간, 0~50℃에 30분 동안 두었을 경우 활성의 변화를 나타내지 않았다.
Production and some properties of chitinolytic enzymes were investigated by 80% ammonium sulfate precipitates (crude enzymes) from culture supernatant of antagonistic bacteria, Chromobacterium violaceum strain C-61 and strain C-72, Aeromonas hydrophila, Aeromonas caviae, and Serratia marcescens, The maximum production of chitinase was obtained from the 3-day culture at 28℃ in C. violaceum strains, the 6-day culture in S. marcescens, and the 2-day culture in A. hydrophila and A. caviae. In the optimum culture periods, chitinase activity of C. violaceum strain C-61 was 1.5, 5.5, 12.0 and 11.3 times higher than those of strain C-72, S. marcescens, A. hydrophila and A. caviae, respectively. However, N,N'-diacetylchitobiase activity was 3.2 times higher in S. marcescens than in C. violaceum strain C-61, and that of Aeromonas spp. was very low. On gels containing glycol chitin, chitinase of C. violaceum strains showed four isoforms of 54-, 52-, 50- and 37-kDa, whereas there were four isoforms of 58-, 52-, 48- and 38-kDa in S. marcescens, three isoforms of 70-, 58- and 54-kDa in A. hydrophila and six isoforms of 90-, 79-, 71-, 63-, 58- and 38-kDa in A. caviae. The chitinase of C. violaceum strain C-61 was most active at pH 7.0 and at 50℃ and was stable in ranges of pH 5.0-10.0 for 2 hours and of 0~50℃ for 30 min.