• 미생물학회지
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• 제49권1호
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• pp.95-98
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• 2013

본 연구에는 대장균에서의 과발현 벡터 개발과 FPLC를 사용한 2단계 컬럼 정제과정을 통해 Pseudomonas alkylphenolia의 alkylphenol hydroxylase의 oxygenase 단백질을 다량으로 정제하는 방법을 개발하였다. 재조합 Escherichia coli BL21(DE3)(pJJPMO2)의 50 g의 wet cake로부터 110 mg의 heterodimer이며 화학량론적 철을 갖는 순수한 단백질을 정제하였으며 147nmole/min/mg의 비활성을 보였다.

• Journal of Microbiology and Biotechnology
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• 제21권11호
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• pp.1179-1183
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• 2011

In this study, the chromosome-encoded pcuRCAXB genes that are required for p-cresol degradation have been identified by using a newly constructed green fluorescent protein (GFP)-based promoter probe transposon in the long-chain alkylphenol degrader Pseudomonas alkylphenolia. The deduced amino acid sequences of the genes showed the highest identities at the levels of 65-93% compared with those in the databases. The transposon was identified to be inserted in the pcuA gene, with the promoterless gfp gene being under the control of the pcu catabolic gene promoter. The expression of GFP was positively induced by p-cresol and was about 10 times higher by cells grown on agar than those in liquid culture. In addition, p-hydroxybenzoic acid was detected during p-cresol degradation. These results indicate that P. alkylphenolia additionally possesses a protocatechuate ortho-cleavage route for p-cresol degradation that is dominantly expressed in colonies.