Abstract
Cultural conditions for the production of extracellar $\beta$-galactosidase by Loctobacillus sporogenes, a spore forming lactic acid bacterium, were investigated with shaken flask and jar fermenter cultures. The fermentation medium giving maximum $\beta$-galactosidase yield was found to consist of 1 % lactose as a carbon source, 1.5% peptone as an organic nitrogen source. 0.2% ammonium sulfate as an inorganic nitrogen source, 0.8% ammonium phosphate dibasic as a phosphorus source, and 0.05% potassium chloride and 0.001% ferric chloride as mineral source. Optimal initial pH of the medium was 7.0 and the highest enzyme excretion was observed after 40 hours of cultivation at 37$^{\circ}C$. In this experiment, the 500$m\ell$ conical flask containing 50-200$m\ell$ of medium was shaken at 140 strokes per minute with 7cm amplitude in a reciprocating shaker. The maximum enzyme value attained was 38 U/$m\ell$ of the culture broth which was found to be slightly higher than the highest intermolecular enzyme activity (30 U/$m\ell$) observed after 24 hours of incubation. In the fermentor culture, the fermentation profile was shown to be similar to that observed in the shaken flask experiment. But the maximum extracellular enzyme activity was 45 U/$m\ell$ to be even higher than the value obtained with the shaken flask culture.
유포자 유산균인 Lactobacillus sporogenes 가 특이하게 균체외 $\beta$-galactosidase를 다량 생산하는 것을 발견하여 효소생산을 위한 영양요구성과 배양조건을 조사하였다. 영양요구성으로는 탄소원으로서 lactose 1%, 유기질소원으로서 peptone 1.5%, 무기질소원으로서 ammonium sulfate 0.2%, phosphorus 원으로 ammonium phosphate, dibasic을 0.8%. mineral은 potassium chloride 0.05%, ferric chloride 0.001% 첨가했을 때 최대의 효소생산을 나타내었다. 배지의 최적 initial pH는 7.0, 최적배양온도는 37$^{\circ}C$, aeration효과는 500$m\ell$용 진탕 flask에 배지량을 50~200$m\ell$ 주입, 140 strokes/min (진폭 7cm)으로 진탕 배양하였을 때 최고의 효소생산을 나타냈다. 상기의 최적조건에서 균체증식은 24시간 배양에 최고에 달하는 반면 $\beta$-galactosidase 생산은 균체내 효소생산의 경우 균체증식 속도와 비례적으로 증가하여 배양 24시간 전후에서 30U/$m\ell$로 최고치를 나타내고 균체외 효소는 배양 40시간 전후 즉, 균증식의 후기 정지기에서 38U/$m\ell$로 가장 높은 효소생산량을 보였다. 또한 fermentor 실험 역시 flask배양과 거의 비슷한 배양 양상을 보였으며 균체외 효소역가는 45U/$m\ell$로 진탕 flask 배양 결과보다 다소 양호하였다.