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An alginate-like exopolysaccharide biosynthesis gene cluster involved in biofilm aerial structure formation by Pseudomonas alkylphenolia

  • Applied microbial and cell physiology
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Abstract

Pseudomonas alkylphenolia is known to form different types of multicellular structures depending on the environmental stimuli. Aerial structures formed during vapor p-cresol utilization are unique. Transposon mutants that showed a smooth colony phenotype failed to form a differentiated biofilm, including aerial structures and pellicles, and showed deficient surface spreading motility. The transposon insertion sites were located to a gene cluster designated epm (extracellular polymer matrix), which comprises 11 ORFs in the same transcriptional orientation. The putative proteins encoded by the genes in the epm cluster showed amino acid sequence homology to those found in the alginate biosynthesis gene clusters, e.g., in Pseudomonas aeruginosa at similarity levels of 32.3–86.4 %. This overall resemblance indicated that the epm gene cluster encodes proteins that mediate the synthesis of an exopolysaccharide composed of uronic acid(s) similar to alginate. Our preliminary results suggested that the epm-derived polymer is a substituted polymannuronic acid. Gene clusters homologous to the epm gene cluster are found in the genomes of a few species of the genera Pseudomonas, Alcanivorax, and Marinobacter. A mutational analysis showed that the epmJ and epmG genes encoding putative exopolysaccharide-modifying enzymes are required to form multicellular structures. An analysis of the activity of the promoter P epmD using a transcriptional fusion to the green fluorescence protein gene showed that the epm genes are strongly expressed at the tips of the specialized aerial structures. Our results suggested that the epm gene cluster is involved in the formation of a scaffold polysaccharide that is required to form multicellular structures in P. alkylphenolia.

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Acknowledgement

This research was financially supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology (no. 2009–0073913 and 2011–0022133) and by the framework of international cooperation program managed by National Research Foundation of Korea (no. 2013K2A1B8053138).

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Authors and Affiliations

  1. Department of Microbiology and Biomedical Science Institute at CWNU, Changwon National University, Changwon-si, Kyongnam, 641-773, South Korea

    Kyoung Lee, Eun Jin Lim & Keun Soo Kim

  2. Department of Life Sciences, National Central University, No. 300 Chung-da Rd., Chung-li, 32001, Taiwan, Republic of China

    Shir-Ly Huang

  3. Amity Institute of Microbial Biotechnology, Amity University, Noida, 201303, India

    Yaligara Veeranagouda

  4. Institute of Fundamental Sciences and MacDiarmid Institute for Advanced Materials and Nanotechnology, Massey University, Private Bag 11222, Palmerston North, New Zealand

    Bernd H. A. Rehm

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  1. Kyoung Lee
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  2. Eun Jin Lim
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Correspondence to Kyoung Lee.

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Lee, K., Lim, E.J., Kim, K.S. et al. An alginate-like exopolysaccharide biosynthesis gene cluster involved in biofilm aerial structure formation by Pseudomonas alkylphenolia . Appl Microbiol Biotechnol 98, 4137–4148 (2014). https://doi.org/10.1007/s00253-014-5529-6

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