Production of Intracellular Invertase from Alkalophilic and Thermophilic Bacillus sp. TA-11 in the Recombinant E. coli

The intracellular invertase gene of alkalophilic and thermophilic Bacillus sp. TA-11 which was isolated from compost was cloned and expressed in E. coil HB101 using pUC19 as a vector. The invertase of the recombinant E. coli (pYC 17) was maximally produced when it was incubated at 37$^{\circ}C$ for 9 h in a SY medium containing 0.25% sucrose, 0.5% yeast extract, 0.1% each of $K_2HPO_4$ and $KH_2PO_4$, with an initial pH of 8.0. The final enzyme activity under the above condition was 47.7 U per ml of cell-free extract.

재조합 대장균에서 호알칼리성,고온성 Bacillus sp. TA-11의 세포내 Invertase의 생산

알칼리와 고온에 대한 내성을 가진 invertase를 대량생산하기 위하여 호알칼리성이며 고온성 세균인 Bacillus sp. TA-11의 세포내 invertase유전자를 pUC 19벡터를 이용하여 E. coli HB101에 클로닝 시키고 발현시켜 invertase를 강력하게 생산하는 재조합 E. coli (pYC17)를 얻었다. 재조합 E. coli(pYC17)를 이용한 invertase 생산 최적조건을 검토한 결과 E. coil(pYC17)를 0.25% sucrose, 0.5% yeast extract, 0.1% $K_2HPO_4$와 0.1% $KH_2PO_4$을 함유한 SY배지(초기 pH 9.0)에 접종하여 37$^{\circ}C$에서 9시간 배양하였을 때 친주보다도 많은 47.7 U/ml-cell free extract의 invertase가 생산되었다.