Isolation and Identification of Microorganism Producing Glutary 7-Aminodeacetoxycephalosporanis Acid Acylase

Glutary 7-Aminodeacetoxycephalosporanis Acid Acylase 생산균의 분리 및 동정

Microorganism producing glutaryl 7-aminodeacetoxycephalosporanic acid (GL-7-ADCA) acylase was screened from soil. The microorganism was identified as Alcaligenes sp. J-421 by its morphology and biochemical properties. Cultural conditions of Alcaligenes sp. J-421 were investigated for the production of GL-7-ADCA acylase. Optimum medium composition was 1% glucose, 1% beef extract, 0.5% yeast extract, 0.2% monosodium L-glutamate, 0.1% glutaric acid, 0.2% NaCl, 0.5% $K_2$ $HPO_4$, and 0.05% $CuSO _4{\cdot}5H_2O$. Optimum cultivation conditions for the production of the enzyme in 5 l jar fermentor were $37^{\circ}C$, tip speed 300 rpm, aeration 1 vvm. Optimum reaction pH of the enzyme was 8.0 and the enzyme was stable at pH7.0-11.0.

Glutaryl 7-aminodeacetoxycephalosporanic acid(GL-7-ADCA)로부터 7-aminodeacetoxycephalosporanic acid(7-ADCA)를 생성하는 GL-7-ADCA acylase생산균을 자연계에서 폭넓게 검색하여 한균주를 선정하고 분리균의 형태 및 생리적 성질들을 조사하여 Alcaligenes sp.로 동정하였다. 분리균의 효소생산을 위한 배양조건을 검토한 결과 탄소원으로는 glucose, 질소원으로는 yeast extract, monosodium L-glutamate가 우수하였다. 효소의 작용최적 pH는 8.0이며, pH7.0~pH11.0 번위에서 비교적 안정하였다. 5l jar fermentor에서 배양경과를 조사한 결과 효소이 활성은 $37^{\circ}C$, 교반속도 300rpm, 통기량 1vvm 조건에서 20시간~30시간 배양시 가장 높았다.