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발색법에 의한 Peroxidase의 신속한 스크리닝법과 2, 4-DCP 분해균주의 스크리닝

Title
발색법에 의한 Peroxidase의 신속한 스크리닝법과 2, 4-DCP 분해균주의 스크리닝
Other Titles
Rapid Screening Method of Peroxidase by Colorimetric Assay and Screening of 2, 4-DCP Degradable Strains
Author
이은규
Keywords
Chlorinated phenols; peroxidase; colorimetric assay; rapid screening; 2,4-DCP degradation strain
Issue Date
2008-12
Publisher
한국생물공학회
Citation
KSBB Journal, v. 23, No. 6, Page. 484-488
Abstract
본 연구에서는 peroxidase 활성의 신속한 스크리닝을 위해 peroxidase의 활성을 정량적으로 평가할 수 있는 분석으로 agar plate와 96-well plate에서 peroxdase의 활성에 비례하여 색상감도를 측정할 수 있는 colorimetric 분석법을 개발하였다. 이 방법은 RP-HPLC 결과와 매우 비례적으로 상관적인 결과를 보였다. 이 colorimetric 분석법을 사용하여, 폐수에서 한천고체배지에서 7회에 걸친 스크리닝으로 높은 농도의 2,4-DCP(1000 ppm)에서 생존하는 균주를 스크리닝하였고 선별된 이들 균주들은 탄소원으로 2,4-DCP 만을 대사할 수 있으며 높은 peroxidase 생산량을 보였다. 분리된 균주들 중 높은 peroxidase 활성을 가지는 2,4-DCP 분해 균주를 분리하였고 최종 분리된 균주는 염색폐수의 COD를 4시간 동안 44%에서 61%까지 제거하였다. 상기의 결과에 의해 본 연구에서 개발된 발색법이 페놀화합물 분해 균주 스크리닝법이 빠르고 쉬운 난분해성 물질인 페놀 혼합물의 분해 균주 탐색에 대하여 성공적으로 적용될 수 있기를 기대한다. Chlorinated phenols are widely used by the chemical industry as intermediate products in synthesis and previously were frequently applied to various industry fields. Peroxidases catalyze the peroxide-dependent oxidation of a range of inorganic and organic compounds. Peroxidase was shown to mineralize a variety of recalcitrant aromatic compounds and to oxidize a number of polycyclic aromatic and phenolic compounds. Among monomeric phenolic and nonphenolic compounds, peroxidase is known to oxidize its compounds. In this study, a colorimetric assay was developed to quantitatively evaluate the peroxidase activity for rapid screening. Color products of different intensity were developed proportionally to the peroxidase activity on agar plate and 96-well plate. This method correlates well with the RP-HPLC result. Using this screening method, 12 colonies of strain was screened which survived at high concentration of 2,4-DCP (1000 ppm) and with peroxidase activity for the 7th round screening step on agar plate. These strains were utilized 2,4-DCP as a sole carbon source and produced peroxidase. After the screening test, four of the bacteria have significant better effect of COD removal on dye waste-water. COD removal of these was from 44% to 61%, respectively.
URI
https://www.earticle.net/Article/A102648https://repository.hanyang.ac.kr/handle/20.500.11754/105294
ISSN
1225-7117
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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