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 |
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Authors: | Faizal Irvan Dozen Kana Hong Chang Soo Kuroda Akio Takiguchi Noboru Ohtake Hisao Takeda Koji Tsunekawa Hiroshi Kato Junichi |
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Affiliation: | 1.Department of Molecular Biotechnology, Hiroshima University, Higashi-Hiroshima, Hiroshima, 739-8530, Japan ;2.Department of Biotechnology, Osaka University, Suita, Osaka, 565-0871, Japan ;3.Mercian Co., Iwata, Shizuoka, 438-0078, Japan ; |
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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. |
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