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Metabolic regulation and chromosomal localization of carbaryl degradation pathway in Pseudomonas sp. strains C4, C5 and C6

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Abstract

Pseudomonas sp. strains C4, C5 and C6 degrade carbaryl (1-naphthyl N-methylcarbamate) via 1-naphthol, 1,2-dihydroxynaphthalene, salicylate and gentisate. Carbon source-dependent metabolic studies suggest that enzymes responsible for carbaryl degradation are probably organized into ‘upper’ (carbaryl to salicylate), ‘middle’ (salicylate to gentisate) and ‘lower’ (gentisate to TCA cycle) pathway. Carbaryl and 1-naphthol were found to induce all carbaryl pathway enzymes, while salicylate and gentisate induce middle and lower pathway enzymes. The strains were found to harbor plasmid(s), and carbaryl degradation property was found to be stable. Genes encoding enzymes of the degradative pathway such as 1-naphthol 2-hydroxylase, salicylaldehyde dehydrogenase, salicylate 5-hydroxylase and gentisate 1,2-dioxygenase were amplified from chromosomal DNA of these strains. The gene-specific PCR products were sequenced from strain C6, and phylogenetic tree was constructed. Southern hybridization and PCR analysis using gel eluted DNA as template supported the presence of pathway genes onto the chromosome and not on the plasmid(s).

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Abbreviations

MSM:

Minimal salt medium

CH:

Carbaryl hydrolase

1NH:

1-Naphthol 2-hydroxylase

12DHNDO:

1,2-Dihydroxynaphthalene dioxygenase

HBHA:

trans-o-Hydroxybenzylidenepyruvate hydratase–aldolase

SALDH:

Salicylaldehyde dehydrogenase

S5H:

Salicylate 5-hydroxylase

GDO:

Gentisate 1,2-dioxygenase

CDO:

Catechol dioxygenase

C12DO:

Catechol 1,2-dioxygenase

C23DO:

Catechol 2,3-dioxygenase

S1H:

Salicylate 1-hydroxylase

ANDO:

Anthranilate 1,2-dioxygenase

PPAD:

3-Phenylpropionate/cinnamic acid dioxygenase

RHO:

Ring-hydroxylating oxygenases

BPDO:

Biphenyl 2,3-dioxygenase

PPDO:

Phenyl propionate dioxygenase

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Acknowledgments

We acknowledge the research fellowship to RS and VDT from CSIR, Govt. of India and the research grant to PP from DBT, Govt. of India. Thanks to Dr. Smalla K, Institute for biochemistry and plant virology, Braunschweig, Germany, for providing standard plasmids for the incompatibility group classification.

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  1. Department of Biosciences and Bioengineering, Indian Institute of Technology-Bombay, Powai, 400 076, Mumbai, India

    Randhir Singh, Vikas D. Trivedi & Prashant S. Phale

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  1. Randhir Singh
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  2. Vikas D. Trivedi
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  3. Prashant S. Phale
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Correspondence to Prashant S. Phale.

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Communicated by Erko Stackebrandt.

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Singh, R., Trivedi, V.D. & Phale, P.S. Metabolic regulation and chromosomal localization of carbaryl degradation pathway in Pseudomonas sp. strains C4, C5 and C6. Arch Microbiol 195, 521–535 (2013). https://doi.org/10.1007/s00203-013-0903-9

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  • DOI: https://doi.org/10.1007/s00203-013-0903-9

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