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Cloning and expression of three ladA-type alkane monooxygenase genes from an extremely thermophilic alkane-degrading bacterium Geobacillus thermoleovorans B23

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Abstract

An extremely thermophilic bacterium, Geobacillus thermoleovorans B23, is capable of degrading a broad range of alkanes (with carbon chain lengths ranging between C11 and C32) at 70 °C. Whole-genome sequence analysis revealed that unlike most alkane-degrading bacteria, strain B23 does not possess an alkB-type alkane monooxygenase gene. Instead, it possesses a cluster of three ladA-type genes, ladAαB23, ladAβB23, and ladB B23, on its chromosome, whose protein products share significant amino acid sequence identities, 49.8, 34.4, and 22.7 %, respectively, with that of ladA alkane monooxygenase gene found on a plasmid of Geobacillus thermodetrificans NG 80-2. Each of the three genes, ladAαB23, ladAβB23, and ladB B23, was heterologously expressed individually in an alkB1 deletion mutant strain, Pseudomonas fluorescens KOB2Δ1. It was found that all three genes were functional in P. fluorescens KOB2Δ1, and partially restored alkane degradation activity. In this study, we suggest that G. thermoleovorans B23 utilizes multiple LadA-type alkane monooxygenases for the degradation of a broad range of alkanes.

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Abbreviations

C12:

Dodecane

C16:

Hexadecane

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Acknowledgments

Pseudomonas fluorescens KOB2Δ1 and pCom8 were kind gifts from Dr. Theo H. M. Smits (Agroscope, Wädenswil, Switzerland). This work was supported by KAKENHI (19380189) to M.M.

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Authors and Affiliations

  1. Division of Biosphere Science, Graduate School of Environmental Science, Hokkaido University, Sapporo, 060-0810, Japan

    Chanita Boonmak, Yasunori Takahashi & Masaaki Morikawa

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  1. Chanita Boonmak
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  2. Yasunori Takahashi
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  3. Masaaki Morikawa
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Correspondence to Masaaki Morikawa.

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Communicated by L. Huang.

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Boonmak, C., Takahashi, Y. & Morikawa, M. Cloning and expression of three ladA-type alkane monooxygenase genes from an extremely thermophilic alkane-degrading bacterium Geobacillus thermoleovorans B23. Extremophiles 18, 515–523 (2014). https://doi.org/10.1007/s00792-014-0636-y

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