Mutational analysis of pcpA and its role in pentachlorophenol degradation by Sphingomonas (Flavobacterium) chlorophenolica ATCC 39723.

نویسندگان

  • S Chanama
  • R L Crawford
چکیده

Sphingomonas (Flavobacterium) chlorophenolica ATCC 39723 degrades pentachlorophenol (PCP) through a catabolic pathway encoded by multiple genes. One gene required for PCP degradation is pcpA, which encodes information for a 30-kDa polypeptide, PcpA, found in the periplasm of the bacterium. The biological role of PcpA has remained unknown. We disrupted pcpA by replacing it with a defective copy through homologous recombination. The pcpA recombinant, mutant strains accumulated 2,6-dichlorohydroquinone (2,6-DiCH) as a metabolite of PCP. This work confirms that pcpA is essential for degradation of PCP by S. chlorophenolica ATCC 39723 and suggests that it encodes a protein involved in hydrolytic dehalogenation of 2,6-DiCH, an already established primary metabolite of the PCP catabolic pathway.

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منابع مشابه

Organization and regulation of pentachlorophenol-degrading genes in Sphingobium chlorophenolicum ATCC 39723.

The first three enzymes of the pentachlorophenol (PCP) degradation pathway in Sphingobium chlorophenolicum (formerly Sphingomonas chlorophenolica) ATCC 39723 have been characterized, and the corresponding genes, pcpA, pcpB, and pcpC, have been individually cloned and sequenced. To search for new genes involved in PCP degradation and map the physical locations of the pcp genes, a 24-kb fragment ...

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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 treatme...

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Phosphorus-31 nuclear magnetic resonance study of the effect of pentachlorophenol (PCP) on the physiologies of PCP-degrading microorganisms.

Free and agarose-encapsulated pentachlorophenol (PCP)-degrading Sphingomonas sp. isolates UG25 and UG30 were compared to Sphingomonas chlorophenolica ATCC 39723 with respect to the ability to degrade PCP. Pretreatment of the UG25 and UG30 strains with 50 microg of PCP per ml enabled the cells to subsequently degrade higher levels of this environmental pollutant. Similar treatment of ATCC 39723 ...

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Cloning, sequence analysis, and expression of the Flavobacterium pentachlorophenol-4-monooxygenase gene in Escherichia coli.

The pcpB gene of Flavobacterium sp. strain ATCC 39723 was cloned by using a degenerate primer designed from the N-terminal sequence of the purified enzyme. The nucleotide sequence of pcpB was determined and found to encode an open reading frame of 1,614 nucleotides, yielding a predicted translation product of 538 amino acids, in agreement with the estimated size of the purified protein analyzed...

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عنوان ژورنال:
  • Applied and environmental microbiology

دوره 63 12  شماره 

صفحات  -

تاریخ انتشار 1997