Enhanced 3-methylcatechol production by Pseudomonas putida TODE1 in a two-phase biotransformation system

The Journal of General and Applied Microbiology

Alisa S. Vangnai¹  Ajiraporn Kongpol³  Takahisa Tajima²  Thunyarat Pongtharangkul⁴  Junichi Kato² 

[1] Department of Biochemistry, Faculty of Science, Chulalongkorn University;Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University;Graduate Program in Biotechnology, Faculty of Science, Chulalongkorn University;Department of Biotechnology, Mahidol University

关键词: an aqueous-organic two-phase fermentation;    bioproduction;    3-methylcatechol;    Pseudomonas putida TODE1;    toluene;    toluene dioxygenase;   

DOI  :  10.2323/jgam.60.183

学科分类：微生物学和免疫学

来源: Applied Microbiology, Molecular and Cellulrar Biosciences Research Foundation

【 摘 要 】

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.

【 授权许可】

Unknown