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Aspergillus niger에서 추출되는 포도당 산화효소의 정제
Purification Study of Glucose Oxidase from Aspergillus niger
충남대학교 화학공학과, 대전 305-764 1RMC Korea, 대전 305-701
Department of Chemical Engineering, Chungnam National University, Daejeon 305-764, Korea 1RMC Korea, TIC/TBI Center in KAIST, Daejeon 305-701, Korea
ihkim@hanbat.chungnam.ac.kr
HWAHAK KONGHAK, August 2001, 39(4), 512-516(5), NONE
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Abstract
Glucose oxidase를 보다 간단한 방법으로 정제하는 방법을 찾기 위해 Table 1에 근거한 여러 가지 정제법을 도입하여 각 단계를 거칠 때마다 단백질의 농도와 GOD의 역가를 측정하였다. Aspergillus niger를 배양한 후 그 상등액을 원심분리하고 전처리 공정으로 증류수를 이용하여 투석을 시킴으로써 시료의 탁도를 떨어뜨리고 음이온 교환기인 DEAE-Sepharose에서 정제한다. 용리액 조성으로 0.1 M NaCl과 0.1M McIlvane buffer(pH 6.0)를 이용하여 GOD를 분리한다. 이러한 정제 과정을 거치게 되면 높은 Sp. activity(304 μmol/mg·min)와 높은 순도(95%)의 glucose oxidase(GOD)을 얻을 수 있었다. 얻어진 GOD는 보고된 GOD의 분자량 150,000과는 달리 그 분자량이 70,000-81,000 사이로 확인되었다.
Glucose oxidase(GOD) from Aspergillus niger was mainly purified by anion exchange chromatography on DEAE-sepharose. Other purification steps such as salting out, cation exchange chromatography, and dialysis were also tried. The purified enzyme was homogeneous on HPLC and SDS-PAGE. We determined an optimum purification condition for high GOD purity. Activity(304 mmol/mg · min), concentration(5.07 mg/mL) and yield (95%) from the starting enzyme source were obtained.
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References
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Ye WN, Combes D, Biochim. Biophys. Acta, 999, 86 (1989)
