Aspergillus niger의 acid phosphatase(EC 3.1.3.2) 는 무기인산 억제 효소이다. 즉 배지에 무기인산이 풍부할 땐 acid phosphatase가 매우 적게 생성되고, 무기인산 결핍 배지일때 abid phosphatase의 생성은 급격히 증가한다. 배지 100ml당 무기인산 10mg일때가 효소생성이 최대였으며, 무기인산의 양이 60mg일때의 약20배 정도 생성되었다. 배지 100ml당 무기인산 10mg인 조건하에서 배양한 균사로 부터 acid phosphatase를 추출하였으며 DEAE-cellulose이온 교환수지 크로마토그라피에 의하여 I과 II로 분리시켰고 Sephadex G-150에 의하여 더 정제하였다. 위의 방법에 의해 각각 40배와 15배 정도 정제되었다. 이렇게 정제된 효소의 p-nitrophenylphosphate(...
Aspergillus niger의 acid phosphatase(EC 3.1.3.2) 는 무기인산 억제 효소이다. 즉 배지에 무기인산이 풍부할 땐 acid phosphatase가 매우 적게 생성되고, 무기인산 결핍 배지일때 abid phosphatase의 생성은 급격히 증가한다. 배지 100ml당 무기인산 10mg일때가 효소생성이 최대였으며, 무기인산의 양이 60mg일때의 약20배 정도 생성되었다. 배지 100ml당 무기인산 10mg인 조건하에서 배양한 균사로 부터 acid phosphatase를 추출하였으며 DEAE-cellulose이온 교환수지 크로마토그라피에 의하여 I과 II로 분리시켰고 Sephadex G-150에 의하여 더 정제하였다. 위의 방법에 의해 각각 40배와 15배 정도 정제되었다. 이렇게 정제된 효소의 p-nitrophenylphosphate(NPP)에 대한 비활성도는 각각 1300과 463이었고 Km값은 각각 $5.3 \times 10^{-4} M$과 $2.5 \times 10^{-4}M$이었다. 이 효소의 최적 pH 는 각각 5.0과 2.0이고 효소 활성을위해 금속 이온을 필요로 하지 않으며 무기인산과 불소이온에 대해 competitive inhibition을 받는다. Acid phosphatase I은 NPP,phosphoenolpyruvate,5'-AMP, glucose 6-phosphate를, acid phosphatase II는 NPP, phosphoenolpyruvate,glucose 6-phosphate,pyridoxal 5'-phosphate 를 잘 가수분해 시키는 반면 ATP,ADT,Adenosine 3'5'-(또는3',2'-) cyclic phosphate는 거의 가수분해하지 않는다. Acid phosphatase I과 II의 분자량은 각각 200,000과 300,000이었다. 그리고 이 효소의 기능은 아직 확실하지 않다.
Aspergillus niger의 acid phosphatase(EC 3.1.3.2) 는 무기인산 억제 효소이다. 즉 배지에 무기인산이 풍부할 땐 acid phosphatase가 매우 적게 생성되고, 무기인산 결핍 배지일때 abid phosphatase의 생성은 급격히 증가한다. 배지 100ml당 무기인산 10mg일때가 효소생성이 최대였으며, 무기인산의 양이 60mg일때의 약20배 정도 생성되었다. 배지 100ml당 무기인산 10mg인 조건하에서 배양한 균사로 부터 acid phosphatase를 추출하였으며 DEAE-cellulose이온 교환수지 크로마토그라피에 의하여 I과 II로 분리시켰고 Sephadex G-150에 의하여 더 정제하였다. 위의 방법에 의해 각각 40배와 15배 정도 정제되었다. 이렇게 정제된 효소의 p-nitrophenylphosphate(NPP)에 대한 비활성도는 각각 1300과 463이었고 Km값은 각각 $5.3 \times 10^{-4} M$과 $2.5 \times 10^{-4}M$이었다. 이 효소의 최적 pH 는 각각 5.0과 2.0이고 효소 활성을위해 금속 이온을 필요로 하지 않으며 무기인산과 불소이온에 대해 competitive inhibition을 받는다. Acid phosphatase I은 NPP,phosphoenolpyruvate,5'-AMP, glucose 6-phosphate를, acid phosphatase II는 NPP, phosphoenolpyruvate,glucose 6-phosphate,pyridoxal 5'-phosphate 를 잘 가수분해 시키는 반면 ATP,ADT,Adenosine 3'5'-(또는3',2'-) cyclic phosphate는 거의 가수분해하지 않는다. Acid phosphatase I과 II의 분자량은 각각 200,000과 300,000이었다. 그리고 이 효소의 기능은 아직 확실하지 않다.
The acid phosphatase(EC 3.1.3.2) of $\mbox{\underline{Aspergillus}}$ $\mbox{\underline{niger}}$ was an orthophosphate-repressible enzyme; in a high phosphate medium, little acid phosphatase was produced, and in a low phosphate medium, the production of acid phosphatase was increased. Medium containi...
The acid phosphatase(EC 3.1.3.2) of $\mbox{\underline{Aspergillus}}$ $\mbox{\underline{niger}}$ was an orthophosphate-repressible enzyme; in a high phosphate medium, little acid phosphatase was produced, and in a low phosphate medium, the production of acid phosphatase was increased. Medium containing 10mg of phosphat per 100ml was optimal, and the amount of acid phosphatase produced in this medium was about 20times of one produced in the medium containing 60mg of phosphate per 10ml. Acid phosphatase was extracted from mycelium of $\mbox{\underline{Aspergillus}}$ $\mbox{\underline{niger}}$ which was grown in the 10mg phosphate per 100ml medium. This enzyme was separated into two fractions(acid phosphatase I and II) by DEAE-cellulose ion-exchange chromatography and further purified by Sephadex G-150 gel filtration. They were purified about $40^-$ and 15-fold, respectively. The partially purified enzymes hydrolyzed 1300, and 463 umoles of p-nitrophenylophosphate (NPP) per minute per mg protein and Km values for NPP were $5.3\times10^{-4}$M and $2.5\times10^{-4}$M, respectively. The activity of acid phosphatase I and II were optimal at $p^H$ 5.0 and 2.0. They did not require the presence of divalent cations. Inorganic orthophosphate and potassium fluoride competitively inhibited hydrolysis of NPP. Acid phosphatase I hydrolyzed NPP, phosphoenolpyruvate, 5'AMP, glucose 6-phosphate and acid phosphatase II hydrolyzed phosphate at maximal rates, while having very little activity for ATP, ADP, and adenosine cyclicphosphate. The molecular weight of partially purified acid phosphatase I and II were estimated to be 200,000 and 300,000 respectively.
The acid phosphatase(EC 3.1.3.2) of $\mbox{\underline{Aspergillus}}$ $\mbox{\underline{niger}}$ was an orthophosphate-repressible enzyme; in a high phosphate medium, little acid phosphatase was produced, and in a low phosphate medium, the production of acid phosphatase was increased. Medium containing 10mg of phosphat per 100ml was optimal, and the amount of acid phosphatase produced in this medium was about 20times of one produced in the medium containing 60mg of phosphate per 10ml. Acid phosphatase was extracted from mycelium of $\mbox{\underline{Aspergillus}}$ $\mbox{\underline{niger}}$ which was grown in the 10mg phosphate per 100ml medium. This enzyme was separated into two fractions(acid phosphatase I and II) by DEAE-cellulose ion-exchange chromatography and further purified by Sephadex G-150 gel filtration. They were purified about $40^-$ and 15-fold, respectively. The partially purified enzymes hydrolyzed 1300, and 463 umoles of p-nitrophenylophosphate (NPP) per minute per mg protein and Km values for NPP were $5.3\times10^{-4}$M and $2.5\times10^{-4}$M, respectively. The activity of acid phosphatase I and II were optimal at $p^H$ 5.0 and 2.0. They did not require the presence of divalent cations. Inorganic orthophosphate and potassium fluoride competitively inhibited hydrolysis of NPP. Acid phosphatase I hydrolyzed NPP, phosphoenolpyruvate, 5'AMP, glucose 6-phosphate and acid phosphatase II hydrolyzed phosphate at maximal rates, while having very little activity for ATP, ADP, and adenosine cyclicphosphate. The molecular weight of partially purified acid phosphatase I and II were estimated to be 200,000 and 300,000 respectively.
주제어
#Phosphatases ProteinsSeperation Aspergillus Aspergillus속 포스파타아제 단백질 분리
학위논문 정보
저자
Ryu, Yeon-Woo
학위수여기관
한국과학기술원
학위구분
국내석사
학과
생물공학과
발행연도
1976
총페이지
v, 64 p.
키워드
Phosphatases ProteinsSeperation Aspergillus Aspergillus속 포스파타아제 단백질 분리
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