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PMID: 9150211 Published · ppublish English Journal Article

p-Cymene catabolic pathway in Pseudomonas putida F1: cloning and characterization of DNA encoding conversion of p-cymene to p-cumate.

Journal of bacteriology ·Vol. 179 ·No. 10 ·1997-05-00 ·Pages 3171-80

Eaton RW

Abstract

Pseudomonas putida F1 utilizes p-cymene (p-isopropyltoluene) by an 11-step pathway through p-cumate (p-isopropylbenzoate) to isobutyrate, pyruvate, and acetyl coenzyme A. The cym operon, encoding the conversion of p-cymene to p-cumate, is located just upstream of the cmt operon, which encodes the further catabolism of p-cumate and is located, in turn, upstream of the tod (toluene catabolism) operon in P. putida F1. The sequences of an 11,236-bp DNA segment carrying the cym operon and a 915-bp DNA segment completing the sequence of the 2,673-bp DNA segment separating the cmt and tod operons have been determined and are discussed here. The cym operon contains six genes in the order cymBCAaAbDE. The gene products have been identified both by functional assays and by comparing deduced amino acid sequences to published sequences. Thus, cymAa and cymAb encode the two components of p-cymene monooxygenase, a hydroxylase and a reductase, respectively; cymB encodes p-cumic alcohol dehydrogenase; cymC encodes p-cumic aldehyde dehydrogenase; cymD encodes a putative outer membrane protein related to gene products of other aromatic hydrocarbon catabolic operons, but having an unknown function in p-cymene catabolism; and cymE encodes an acetyl coenzyme A synthetase whose role in this pathway is also unknown. Upstream of the cym operon is a regulatory gene, cymR. By using recombinant bacteria carrying either the operator-promoter region of the cym operon or the cmt operon upstream of genes encoding readily assayed enzymes, in the presence or absence of cymR, it was demonstrated that cymR encodes a repressor which controls expression of both the cym and cmt operons and is inducible by p-cumate but not p-cymene. Short (less than 350 bp) homologous DNA segments that are located upstream of cymR and between the cmt and tod operons may have been involved in recombination events that led to the current arrangement of cym, cmt, and tod genes in P. putida F1.

MeSH Terms
Acetate-CoA Ligase/metabolism Alcohol Dehydrogenase/metabolism Aldehyde Dehydrogenase/metabolism Amino Acid Sequence Bacterial Outer Membrane Proteins/metabolism Base Sequence Benzoates/metabolism Biotransformation/genetics Cloning, Molecular Cymenes DNA, Bacterial/chemistry,genetics,metabolism Enoyl-CoA Hydratase/genetics Genes, Bacterial Mixed Function Oxygenases/metabolism Molecular Sequence Data Monoterpenes Operon Oxidoreductases/metabolism Oxo-Acid-Lyases/genetics Pseudomonas putida/enzymology,genetics,metabolism Repetitive Sequences, Nucleic Acid Terpenes/metabolism Toluene/metabolism
Chemicals
Bacterial Outer Membrane Proteins Benzoates Cymenes DNA, Bacterial Monoterpenes Terpenes 4-cymene Toluene cumic acid Mixed Function Oxygenases Oxidoreductases Alcohol Dehydrogenase Aldehyde Dehydrogenase 4-hydroxy-2-oxovalerate aldolase Oxo-Acid-Lyases Enoyl-CoA Hydratase Acetate-CoA Ligase
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Eaton R W
National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Gulf Breeze, Florida 32561, USA. [email protected]
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.10.3171-3180.1997
Email
[email protected]
Published
1997-05-00
Pages
3171-80
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179094
Subset
IM
Databases
GENBANK
U24215
External Links
PubMed source DOI: 10.1128/jb.179.10.3171-3180.1997 PMC Full Text View journal details
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