Microbial Transglutaminase의 비연속 분리공정 최적화

Membranes are widely used to separate target solute molecules such as proteins on the basis of their size in cell broth mixture to minimize the loss of target compounds. In this study, membrane separation system using ultrafilters of MWCO 100 K and 50 K, was operated for concentration and purification of microbial transglutaminase. Fermentation broth containing MTGase was prefiltered by using pore size 1.6 and $0.7\;{\mu}m$ pre-filter made of cellulose fiber and $0.45\;{\mu}m$ microfilter made of cellulose acetate. The prefiltered solution was concentrated by 100 K and 50 K ultrafilter. The final enzyme concentration was 1.29 units/ml and enzyme specific activity was 0.2 units/mg protein. This specific activity were 3.7 times higher than that of initial cell broth mixture. Membrane separation process of microfiltration and ultrafiltration was proved to be very economic, energy efficient and effective separation method used to concentrate MTGase.

대량발효 후 미세여과막과 MWCO 100K, 50K의 한외여과막을 이용한 막분리시스템을 적용하여 MTGase를 분리한 결과 pore size 1.6, $0.7\;{\mu}m$의 cellulose fiber 재질의 예비여과막과 $0.45\;{\mu}m$ cellulose acetate재질의 미세여과막을 이용하여 얻어진 분리액을 MWCO 100K와 50K를이용하여 농축시킨 결과 enzyme의 농도가 1.29 units/ml, 효소 비활성도는 약 0.2 units/mg protein 으로 나타났으며 초기 배양액에 비해 3.7배의 농축효과를 보였다.