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Molecular cloning, genetic organization of gene cluster encoding phenol hydroxylase and catechol 2,3-dioxygenase in Alcaligenes faecalis IS-46

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Abstract

Alcaligenes faecalis IS-46 can utilize phenol as the sole carbon and energy source at concentration up to 1000 mg/l. In this report we created a cosmid library of this strain and the two clones specifying the whole L-46d type of phenol hydroxylase gene cluster were identified and characterized. Sequence analysis revealed that although the overall gene organization of the clusters was quite similar, few coding sequences differed or were found to have two copies compared with other source organisms. One of these coding sequences showed a good protein sequence similarity to a hypothetical protein and one matched with a regulatory protein of the LysR system. Their putative role in phenol degradation was discussed. Bioinformatic analysis suggested tentative phylogenetic assignments of the retrieved clusters. This work described for first time the complete nucleotide sequence and genetic organization of the whole phenol hydroxylase gene cluster in A. faecalis species.

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Abbreviations

PH:

Phenol hydroxylase

mPH:

Multicomponent phenol hydroxylase

LmPH:

The largest subunit of the multicomponent phenol hydroxylase

C23O:

Catechol 2,3-dioxygenase

CatA:

Catechol 1,2-dioxygenase

2-HMS:

2-Hydroxymuconic semialdehyde

ORF:

Open reading frame

IS:

Insertion sequences

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Acknowledgements

This work was supported by the grants from the National Natural Science Foundation of China (30470061 and 20677041) and the Shanghai Rhone-Alpes bilateral collaboration project of Shanghai Municipal Committee of Science and Technology (05SR07107). The authors thank Dr. He Xinyi and Dr. Zhang Yan for recommendation of gene cloning and screening protocols, Dr. Zhang Xueli (Univ. of Florida, USA) for critical reading of the manuscript.

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  1. Laboratory of Molecular Microbial Ecology and Ecogenomics, College of Life Science and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan road, Shanghai, 200240, China

    Chenguang Zhu, Liye Zhang & Liping Zhao

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  1. Chenguang Zhu
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  2. Liye Zhang
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  3. Liping Zhao
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Correspondence to Liping Zhao.

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Zhu, C., Zhang, L. & Zhao, L. Molecular cloning, genetic organization of gene cluster encoding phenol hydroxylase and catechol 2,3-dioxygenase in Alcaligenes faecalis IS-46. World J Microbiol Biotechnol 24, 1687–1695 (2008). https://doi.org/10.1007/s11274-008-9660-3

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