Studies on Microbial Penicillin Amidase (II) Characteristics and the Reactor Performance of Whole Cell Immobilized Penicillin Amidase of Escherichia coli

미생물 페니실린 아미다제에 관한 연구 (II) E. coli의 균체 고정화 페니실린 아미다제의 특성 및 반응조에 관한 연구

  • Seong, Baik-Lin (Biotechnology Research Department, Korea Institute of Science and Technology) ;
  • Kim, Bong-Hee (Biotechnology Research Department, Korea Institute of Science and Technology) ;
  • Mheen, Tae-Iek (Biotechnology Research Department, Korea Institute of Science and Technology) ;
  • Moon H. Han (Biotechnology Research Department, Korea Institute of Science and Technology)
  • Published : 1981.03.01

Abstract

Whole cell penicillin amidase of Escherichia coli was immobilized by entrapment in gelatin followed by extrusion and crosslinking with glutaraldehyde. The immobilized engyme preparation demonstrated the recovery yield of activity up to 70% and good stability during storage and operation. The half life of activity decay during the operation was estimated to be about 50 days. The optimum pH and temperature for both of immobilized and soluble enzyme are 8.5 and 5$0^{\circ}C$, respectively. No significant change was demonstrated in the effect of pH and temperature, but the increase in heat stability at high temperature was observed in the case of the immobilized enzyme. It was found that the plug flow reactor could be operated favorably since the pH drop along the column path due to tile reaction product was minimized by employing substrate solution with moderate buffer strength. The optimal condition of reactor operation was discussed with regard to the effect of substrate concentration and the residence time on the conversion efficiency and productivity.

대장균이 생산하는 페니실린 아미다제를 젤라틴에 포괄시켜 사출한 후 글루트알데히드로 가교하여 고정화하였다. 이렇게 하여 만들어진 고정화효소는 약 70%의 높은 효소역가를 나타내었고 보관 및 반응조에서 좋은 안정성을 보여주었다. 반응조 내에서의 효소역가 반감기는 약 50일이었으며 최적 PH 및 온도는 각각 8.5와 5$0^{\circ}C$로 나타났다. 효소역가에 미치는 pH 및 온도의 영향은 고정화하기 전과 큰 차이가 없었으나 고온에서의 안정성이 증가되었다. 기질용액으로 완충액을 사용하여 column을 사용하는 관형식 반응조에서의 반응생성률에 기인하는 pH 감소효과를 최소한으로 줄이므로써 효소반응조를 최적화하였다. 반응조 조작상의 중요한 인자 즉 기질농도, 체류시간, 반응 생성물로의 전환율 및 이에 따르는 생산성을 pH 감소효과와 연관시켜 최적반응조건을 논의하였다.

Keywords