Isolation and characterization of solvent-tolerant Pseudomonas putida strain T-57, and its application to biotransformation of toluene to cresol in a two-phase (organic-aqueous) system

  • Irvan Faizal
  • Kana Dozen
  • Chang Soo Hong
  • Akio Kuroda
  • Noboru Takiguchi
  • Hisao Ohtake
  • Koji Takeda
  • Hiroshi Tsunekawa
  • Junichi Kato
Environmental Biotechnology

Abstract

Pseudomonas putida T-57 was isolated from an activated sludge sample after enrichment on mineral salts basal medium with toluene as a sole source of carbon. P. putida T-57 utilizes n-butanol, toluene, styrene, m-xylene, ethylbenzene, n-hexane, and propylbenzene as growth substrates. The strain was able to grow on toluene when liquid toluene was added to mineral salts basal medium at 10–90% (v/v), and was tolerant to organic solvents whose log  P ow (1-octanol/water partition coefficient) was higher than 2.5. Enzymatic and genetic analysis revealed that P. putida T-57 used the toluene dioxygenase pathway to catabolize toluene. A cis-toluene dihydrodiol dehydrogenase gene (todD) mutant of T-57 was constructed using a gene replacement technique. The todD mutant accumulated o-cresol (maximum 1.7 g/L in the aqueous phase) when cultivated in minimal salts basal medium supplemented with 3% (v/v) toluene and 7% (v/v) 1-octanol. Thus, T-57 is thought to be a good candidate host strain for bioconversion of hydrophobic substrates in two-phase (organic-aqueous) systems.

Keywords

Two-phase system Solvent tolerant Toluene dioxygenase pathway Pseudomonas putida Monooxygenation 

Notes

Acknowledgements

This work was carried out as part of the Project for Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers by the Ministry of Economy, Trade, and Industry (METI), which was entrusted by New Energy and Industrial Technology Development Organization (NEDO).

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Copyright information

© Society for Industrial Microbiology 2005

Authors and Affiliations

  • Irvan Faizal
    • 1
  • Kana Dozen
    • 1
  • Chang Soo Hong
    • 1
  • Akio Kuroda
    • 1
  • Noboru Takiguchi
    • 1
  • Hisao Ohtake
    • 2
  • Koji Takeda
    • 3
  • Hiroshi Tsunekawa
    • 3
  • Junichi Kato
    • 1
  1. 1.Department of Molecular BiotechnologyHiroshima UniversityHigashi-HiroshimaJapan
  2. 2.Department of BiotechnologyOsaka UniversitySuitaJapan
  3. 3.Mercian Co.IwataJapan

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