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Genes involved in alkane degradation in the Alcanivorax hongdengensis strain A-11-3

  • Applied genetics and molecular biotechnology
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Abstract

Alcanivorax hongdengensis A-11-3 is a newly identified type strain isolated from the surface water of the Malacca and Singapore Straits that can degrade a wide range of alkanes. To understand the degradation mechanism of this strain, the genes encoding alkane hydroxylases were obtained by PCR screening and shotgun sequencing of a genomic fosmid library. Six genes involved in alkane degradation were found, including alkB1, alkB2, p450-1, p450-2, p450-3 and almA. Heterogeneous expression analysis confirmed their functions as alkane oxidases in Pseudomonas putida GPo12 (pGEc47ΔB) or Pseudomonas fluorescens KOB2Δ1. Q-PCR revealed that the transcription of alkB1 and alkB2 was enhanced in the presence of n-alkanes C12 to C24; three p450 genes were up-regulated by C8–C16 n-alkanes at different levels, whereas enhanced expression of almA was observed when strain A-11-3 grew with long-chain alkanes (C24 to C36). In the case of branched alkanes, pristane significantly enhanced the expression of alkB1, p450-3 and almA. The six genes enable strain A-11-3 to degrade short (C8) to long (C36) alkanes that are straight or branched. The ability of A. hongdengensis A-11-3 to thrive in oil-polluted marine environments may be due to this strain’s multiple systems for alkane degradation and its range of substrates.

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Acknowledgements

We would like to thank Dr. Daniela Näther and Dr. Montri Yasawong (Environmental Microbiology Group, Helmholtz Centre for Infection Research) for their help with genomic fosmid library construction and analysis. We would like to thank Dr. Jan B. van Beilen (Institute of Biotechnology, Zurich, Switzerland) for kindly providing P. fluorescens KOB2D1, P. putida Gpo12, E. coli CC118 (RK600) and plasmids pGEc47DB, pCom8 and pRK2017. This work was financially supported by The Project Sponsored by the National Science Foundation of China (41176154, 41106151), the International Sci & Tech cooperation Program of China (2010DFB93670), and the Sci & Tech Program of Fujian Province of China (2009H0029).

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Authors and Affiliations

  1. Key Laboratory of Marine Biogenetic Resources, The Third Institute of Oceanography, State Oceanic Administration, Daxue Road 178, Xiamen, 361005, China

    Wanpeng Wang & Zongze Shao

  2. School of Life Sciences, Xiamen University, Xiamen, 361005, China

    Wanpeng Wang

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  1. Wanpeng Wang
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  2. Zongze Shao
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Correspondence to Zongze Shao.

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Table S1

List of predicted ORFs retrieved from 826 fosmid clone (DOC 72.5 kb)

Table S2

List of predicted ORFs retrieved from 118 fosmid clone (DOC 81.5 kb)

Table S3

List of predicted ORFs retrieved from 517 fosmid clone (DOC 88 kb)

Table S4

List of predicted ORFs retrieved from 208 fosmid clone (DOC 66.5 kb)

Table S5

List of predicted ORFs retrieved from 336 fosmid clone (DOC 74.5 kb)

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Wang, W., Shao, Z. Genes involved in alkane degradation in the Alcanivorax hongdengensis strain A-11-3. Appl Microbiol Biotechnol 94, 437–448 (2012). https://doi.org/10.1007/s00253-011-3818-x

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  • DOI: https://doi.org/10.1007/s00253-011-3818-x

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