Publication: Enhanced 3-methylcatechol production by pseudomonas putida TODE1 in a two-phase biotransformation system
Issued Date
2014-01-01
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ISSN
13498037
00221260
00221260
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2-s2.0-84924756875
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of General and Applied Microbiology. Vol.60, No.5 (2014), 183-190
Suggested Citation
Ajiraporn Kongpol, Junichi Kato, Takahisa Tajima, Thunyarat Pongtharangkul, Alisa S. Vangnai Enhanced 3-methylcatechol production by pseudomonas putida TODE1 in a two-phase biotransformation system. Journal of General and Applied Microbiology. Vol.60, No.5 (2014), 183-190. doi:10.2323/jgam.60.183 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/34021
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Title
Enhanced 3-methylcatechol production by pseudomonas putida TODE1 in a two-phase biotransformation system
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Abstract
©2014 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation. In this study, genetically engineered Pseudomonas putida TODE1 served as a biocatalyst for the bioproduction of valuable 3-methylcatechol (3MC) from toluene in an aqueous-organic two-phase system. The two-phase system was used as an approach to increase the biocatalyst efficiency. Among the organic solvent tested, n-decanol offered several benefits including having the highest partitioning of 3MC, with a high 3MC yield and low cell toxicity. The effect of media supplementation with carbon/ energy sources (glucose, glycerol, acetate and succinate), divalent metal cations (Mg2+, Ca2+, Mn2+ and Fe2+), and short-chain alcohols (ethanol, n-propanol and n-butanol) as a cofactor regeneration system on the toluene dioxygenase (TDO) activity, cell viability, and overall 3MC yield were evaluated. Along with the two-step cell preparation protocol, supplementation of the medium with 4 mM glycerol as a carbon/energy source, and 0.4 mM Fe2+ as a cofactor for TDO significantly enhanced the 3MC production level. When in combination with the use of n-decanol and n-butanol as the organic phase, a maximum overall 3MC concentration of 31.8 mM (166 mM in the organic phase) was obtained in a small-scale production, while it was at 160.5 mM (333.2 mM in the organic phase) in a 2-L scale. To our knowledge, this is the highest 3MC yield obtained from a TDO-based system so far.