키토산을 효소로 분해하여 생리활성물질이 우수한 chitooligo당을 분리하여 정제를 시도하였다. Chitosan을 탈 아세틸화 시킨 후 탈 아세틸화도(DAC)도가 DAC-45%, DAC-70%, DAC-75% 및 DAC-99%를 기질로 하여 chitosanase(Bacillus pumilus BN-282)로 분해한 결과 DAC-99가 분해능이 가장 우수하였다. 따라서 DAC-90을 이용하여 chitosanase로 분해한 다음 gel filteration하여 Biol-2 gel chromatography로 분획한 획분을 HPLC로 분리 정제한 결과 chitooligo당의 조성은 GlcN, GlcN_2, GlcN_3, GlcN_5 및 GlcN_6임을 확인하였다.
This studies were carried out to purification and separation of chitooligosacchariches which containing excellent biological active substance. After deacetylation of chitosan (DAC%), DAC-45%, DAC-70%, DAC-95% and DAC-99% were used substrates and hydrolyzed by chitosanase (Bacillus pumilus BN-262) DAC-99% has excellent hydrolyzate which contained several chitooligosaccharides. Therfore, chitosan was hydrolyzed DAC-90 as substrate by chitosanase, and then purified and seperated of chitooligosaccharides Gel filteration and HPLC. This oligosaccharides composed with GlcN_0, GlcN_2, GlcN_3, GlcN_5 and GlcN_6.