Purification of a Flavobacterium pentachlorophenol-induced periplasmic protein (PcpA) and nucleotide sequence of the corresponding gene (pcpA).
A pentachlorophenol (PCP)-induced periplasmic protein (PcpA) with a molecular weight of 30,000 has been identified and purified from Flavobacterium sp. strain ATCC 39723. Results suggest that PcpA may be involved in PCP mineralization. The induction of PcpA correlated with the induction of PCP-degrading activity. When PcpA was released from the periplasmic space by EDTA or osmotic shock treatment, PCP-degrading activity was arrested. Two PCP- mutants of ATCC 39723, which do not degrade PCP, did not produce PcpA following induction with PCP. The N terminus of PcpA was sequenced, and two degenerate primers were synthesized for use in DNA amplification of a 68-bp probe which hybridized to a genomic library clone containing pcpA. The nucleotide sequence of pcpA was determined and found to encode an open reading frame of 816 bp. pcpA was found to be part of a 1.35-kb transcript with a transcriptional start site 67 bp upstream of the translational start site. Data base search comparisons yielded no sequences of high similarity to pcpA or its protein product.
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Department of Bacteriology and Biochemistry, University of Idaho, Moscow 83843.
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002268:Carboxypeptidases / Q000235:genetics* / Q000302:isolation & purification; D043422:Carboxypeptidases A; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004492:Edetic Acid / Q000494:pharmacology; D004792:Enzyme Precursors / Q000235:genetics* / Q000302:isolation & purification; D005417:Flavobacterium / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D010416:Pentachlorophenol / Q000378:metabolism; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D014158:Transcription, Genetic
D004792:Enzyme Precursors; D008565:Membrane Proteins; D012313:RNA; D004492:Edetic Acid; D010416:Pentachlorophenol; D002268:Carboxypeptidases; D043422:Carboxypeptidases A