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Volume 149, Issue 3

Expansion of growth substrate range in F1 by mutations in both and , which recruit a ring-fission hydrolase CmtE and induce the catabolic operon, respectively Free

Abstract

F1 can assimilate benzene, toluene and ethylbenzene using the toluene degradation pathway, and can also utilize -cymene via -cumate using the -cymene and -cumate catabolic pathways. In the present study, F1 strains were isolated that were adapted to assimilate new substrates such as -propylbenzene, -butylbenzene, cumene and biphenyl, and the molecular mechanisms of genetic adaptation to an expanded range of aromatic hydrocarbons were determined. Nucleotide sequence analyses showed that the selected strains have mutations in the gene but not in gene. The impairment of the repressor CymR by mutation led to the constitutive expression of CmtE, a -cleavage product hydrolase from the operon. This study also showed that CmtE has a broad range of substrates and can hydrolyse -cleavage products formed from biphenyl and other new growth substrates via the toluene degradation pathway. However, the artificially constructed strain F1( : : Tc) and a recombinant F1, which expressed CmtE constitutively, could not grow on the new substrates. The adapted strains possess the operon, which is induced by new growth substrates that are poor inducers of wild-type F1. When the gene from the adapted strains was introduced in a manner to F1( : : Tc), the resulting recombinant strains were able to grow on biphenyl and other new substrates. This finding indicates that the TodS sensor was altered to recognize these substrates and this conclusion was confirmed by nucleotide sequence analyses. Amino acid substitutions were found in the regions corresponding to the receiver domain and the second PAS domain and their boundaries in the TodS protein. These results showed that F1 adapted strains capable of growth on -propylbenzene, -butylbenzene, cumene and biphenyl possess mutations to employ CmtE and to induce the catabolic operon by the new growth substrates.

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Keyword(s): GFP, green fluorescent protein , HOHD, 2-hydroxy-6-oxohexa-2,4-dienoate , Tc, tetracycline and tod, toluene degradation
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2003-03-01
2024-04-16
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Expansion of growth substrate range in Pseudomonas putida F1 by mutations in both cymR and todS, which recruit a ring-fission hydrolase CmtE and induce the tod catabolic operon, respectively
Microbiology 149, 795 (2003); https://doi.org/10.1099/mic.0.26046-0
/content/journal/micro/10.1099/mic.0.26046-0
/content/journal/micro/10.1099/mic.0.26046-0
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