Simulation and optimization of two-stage continuous stirred tank enzyme reactor system : Penicillin amidohydrolase for production of 6-anmino-penicillanic acid
반합성페니실린의 출발물질인 6-아미노페니실란산 (6-APA)의효소적 제조방법에 있어, Bacillus megaterium(ATCC 14976)균주로부터 얻는 penicillin amidohydrolase (EC 3,5,1,11)의 생산수율이 배지조성의 변화, 효소유발물질 첨가농도의 증가등을 포함한 균체배양 방법의 개량에 의하여 증가되었음을 보고하였다. 또한 효소반응에 있어서 반응 생성물에 의한 경쟁적 및 비경쟁적 2중 저해를 동시에 고려한 ...
반합성페니실린의 출발물질인 6-아미노페니실란산 (6-APA)의효소적 제조방법에 있어, Bacillus megaterium(ATCC 14976)균주로부터 얻는 penicillin amidohydrolase (EC 3,5,1,11)의 생산수율이 배지조성의 변화, 효소유발물질 첨가농도의 증가등을 포함한 균체배양 방법의 개량에 의하여 증가되었음을 보고하였다. 또한 효소반응에 있어서 반응 생성물에 의한 경쟁적 및 비경쟁적 2중 저해를 동시에 고려한 반응속도식을 도출하였고 여기에 2단계 연속교반조반응기의 reactor kinetics 를 더하여서 computer simulation 기법에 의하여 그 반응장치의 최적조작 조건을 찾아 내었다. 그 결과 penicillin amidohydrolase 효소의 고정화 형태를쓰는 것이 수용성 효소를 그대로 사용하는 것보다 동일한 반응조건하에서 훨씬 높은 생산성을 나타냄을 알았으며 또한 2단계 연속 교반조 반응장치를 사용하는 것이 1단계만을 사용하는 것보다도보다 향상된 생산성을 가짐을 알게 되었다.
반합성 페니실린의 출발물질인 6-아미노페니실란산 (6-APA)의효소적 제조방법에 있어, Bacillus megaterium(ATCC 14976)균주로부터 얻는 penicillin amidohydrolase (EC 3,5,1,11)의 생산수율이 배지조성의 변화, 효소유발물질 첨가농도의 증가등을 포함한 균체배양 방법의 개량에 의하여 증가되었음을 보고하였다. 또한 효소반응에 있어서 반응 생성물에 의한 경쟁적 및 비경쟁적 2중 저해를 동시에 고려한 반응속도식을 도출하였고 여기에 2단계 연속교반조반응기의 reactor kinetics 를 더하여서 computer simulation 기법에 의하여 그 반응장치의 최적조작 조건을 찾아 내었다. 그 결과 penicillin amidohydrolase 효소의 고정화 형태를쓰는 것이 수용성 효소를 그대로 사용하는 것보다 동일한 반응조건하에서 훨씬 높은 생산성을 나타냄을 알았으며 또한 2단계 연속 교반조 반응장치를 사용하는 것이 1단계만을 사용하는 것보다도보다 향상된 생산성을 가짐을 알게 되었다.
Penicillin amidohydrolase, an enzyme which hydrolyzes benzylpenicillin to give 6-Aminopenicillanic acid and phenylacetic acid, was produced by using $\mbox{\underline{Bacillus}}$ $\mbox{\underline{megaterium}}$ culture. Production of the enzyme from the strain was increased 20 fold by modification o...
Penicillin amidohydrolase, an enzyme which hydrolyzes benzylpenicillin to give 6-Aminopenicillanic acid and phenylacetic acid, was produced by using $\mbox{\underline{Bacillus}}$ $\mbox{\underline{megaterium}}$ culture. Production of the enzyme from the strain was increased 20 fold by modification of culture medium and the fermentation method. Larger amount of phenyl acetic acid than reported by other investigators was added to the actively growing cell to induce the maximum production of the enzyme. When casitone-containing medium was used, the highest enzyme yield was obtained. Optimal temperature and $P^H$ for the activity of bentonite-immobilized penicillin amidohydrolase was 40-45$^\circ$C and 8-9, respectively. The performance of two-stage continuous stirred tank enzyme reactor (CSTR) system was evaluated from the results of computer simulation study of the system. The productivity of the reactor system using immobilized enzyme was much higher than using soluble enzyme, which was mainly due to the higher value of product inhibition constant of immobillzed enzyme than that of soluble enzyme. The productivity obtained with the two-stage CSTR system was found to be 9.5 m mole of 6-APA per liter of reactor working volume per hour and it was 50\% higher than that obtained with the single stage system under the given operational condition. The optical feed concentration of substrate that give the maximum productivity was found to be 0.18 M for 95\% conversion requirement and corresponding the first and second stage space time were 10 hours and 8 hours, respectively.
Penicillin amidohydrolase, an enzyme which hydrolyzes benzylpenicillin to give 6-Aminopenicillanic acid and phenylacetic acid, was produced by using $\mbox{\underline{Bacillus}}$ $\mbox{\underline{megaterium}}$ culture. Production of the enzyme from the strain was increased 20 fold by modification of culture medium and the fermentation method. Larger amount of phenyl acetic acid than reported by other investigators was added to the actively growing cell to induce the maximum production of the enzyme. When casitone-containing medium was used, the highest enzyme yield was obtained. Optimal temperature and $P^H$ for the activity of bentonite-immobilized penicillin amidohydrolase was 40-45$^\circ$C and 8-9, respectively. The performance of two-stage continuous stirred tank enzyme reactor (CSTR) system was evaluated from the results of computer simulation study of the system. The productivity of the reactor system using immobilized enzyme was much higher than using soluble enzyme, which was mainly due to the higher value of product inhibition constant of immobillzed enzyme than that of soluble enzyme. The productivity obtained with the two-stage CSTR system was found to be 9.5 m mole of 6-APA per liter of reactor working volume per hour and it was 50\% higher than that obtained with the single stage system under the given operational condition. The optical feed concentration of substrate that give the maximum productivity was found to be 0.18 M for 95\% conversion requirement and corresponding the first and second stage space time were 10 hours and 8 hours, respectively.
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