Culture Conditions of E. coli CK1092 for the Production of 2,3-Dihydroxybiphenyl Dioxygenase

2,3-Dihydroxybiphenyl Dioxygenase 생산을 위한 E. coli CK1092의 배양조건

To obtain higher yield of 2,3-dihydroxybiphenyl(2,3-DHBP) dioxygenase by recombinant E. coli CK1092 carrying pcbC gene of Pseudomonas sp. P20, the environmental and physiological factors were investigated and the cultural conditions using jar fermentor were studied. E. coli CKl092 was grown in LB medium supplemented with 2% sucrose, as a basal medium. The effect of various metal ions on the enzyme production was investigated. In particular, the enzyme production increased in the presence of $Fe^{3+}$ and $Fe^{2+}$, and showed the maxium at the concentration of $10^{-5}M$. The enzyme production was increased by 55% in the medium containing $Fe^{3+}$ ($10^{-5}M$) ion. The optimal temperature and initial pH for cell growth were $37^{\circ}C$ and 7.0, respectively. In the culture using a fermentor at $37^{\circ}C$, the optimal conditions for the enzyme production were obtained at the initial pH 7.0, 1 v/v/m of aeration rate, 200 rpm of agitation speed. It was found that enzyme activity was higher when cultivated without pH control than with pH control.

PCBs를 분해하는 Pseudomonas sp. P20 균주의 pcbC gene이 재조합 된 E. coli CK1092 균주를 이용하여 2,3-dihydroxybiphenyl dioxygenase 생산에 미치는 배양조건을 검토하였다. E. coli CK1092를 2% sucrose가 포함된 LB 배지를 기본배지로 하여 질소원, 금속이온 등의 영향을 조사한 결과, $Fe^{3+}$와 $Fe^{2+}$이 $10^{-5}M$의 농도일때 효소생산이 증가되었으며 배양최적온도는 $37^{\circ}C$, 배양초기 pH 7.0일때 효소생산이 우수하였다. 배양조에서의 배양조건은 초기 pH 7.0, 통기량 1 v/v/m, 교반속도 200rpm에서 가장 효소생성이 우수하였다.