Serratia marcescen ATCC 25419 protease를 ammonium sulfate treatment, DEAE-cellulose anion exchange chromatography등의 방법으로 정제하였는데 최종 단계에서 667.5 unit/mg 이었으며 회수율은 43%이었고 448배 정제되었다. 정제한 S. marcescens protease로부터 아포효소를 만든 후 금속 재활성화에 대해 조사하였다. S. marcescens protease는 EDTA에 의해 완전히 활성을 잃는 metalloenzyme이며 Hg, Fe, Cu 등에 의해서 효소 활성을 70% 이상 잃은 반면, Co는 효소 활성을 약 20% 정도 증가시켰다. 아포효소의 재활성화는 pH 6~8에서 Mn, Co, Zn 등이 효과적이었다. Mn, Co, Zn등을 아포효소에 가하여 만든 효소들 중에서 Zn-효소는 효소 활성도, 알칼리-불활성화, 열-안정성 면에서 원래 protease와 유사하였다.
Serratia marcescens ATCC 25419 protease was purified to homogeneity by ammonium sulfate treatment, and DEAE-cellulose anion exchange chromatography. The specific activity of the enzyme was increased 448-fold during purification with an overall yield of 43.0%. Metal reactivation on the purified protease from S. marcescens was studied. S. marcescens protease was a metalloenzyme to be completely inhibited its activity by EDTA and the enzyme outstandingly inhibited by Hg, Fe, Cu, but the activity was increased approximately 20% by Co. The reactivation of the apoenzyme was effective with Mn, Co, Zn in pH range from 6 to 8. Among metalloenzymes prepared to the addition of Mn, Co, Zn to restore the degree of activity of native enzyme, Zn-enzyme was similar to the native enzyme in respects with enzyme activity, alkali-inactivation, thermo-stability.
