Journal
MICROBIOLOGICAL RESEARCH
Volume 165, Issue 8, Pages 649-656Publisher
ELSEVIER GMBH, URBAN & FISCHER VERLAG
DOI: 10.1016/j.micres.2009.11.011
Keywords
Degradation; NADH oxidoreductase; Phenylacetic acid; Streptomyces peucetius
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Funding
- Ministry of Education, Science and Technology [KRF-2008-C00534]
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Annotation of genome of Streptomyces peucetius revealed a putative phenylacetic acid degradation NADH oxidoreductase. RT-PCR analysis of the gene readily showed notable transcription in its native state. The transcription level of paaE when the host is grown on phenylacetic acid showed increased transcription. paaE was cloned into a pET32a(+) vector to overexpress the protein coupled with fusion tags in Escherichia coli BL21(DE3) and purified by immobilized metal affinity chromatography using His-tag. The flavin released from heat-denatured PaaE was identical to that of authentic FAD in HPLC analysis. The purified protein efficiently reduced p-nitroblue tetrazolium (an electron acceptor) in presence of NADH. Cell growth analysis of S. peucetius in phenylacetic acid evidently revealed its involvement in degradation of phenylacetic acid - a key environmental pollutant. (C) 2009 Elsevier GmbH. All rights reserved.
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