Engineering Toluene Monooxygenases for Synthesis of Hydroxytyrosol
The overall goal of this research is to design toluene monooxygenases (TMOs) via protein engineering techniques in order to obtain enzymes with altered regiospecificity, and to study structure-function implications. Furthermore, special attention is given to the biosynthesis of commercially-valuable hydroxytyrosol (HTyr), a potent antioxidant, from an inexpensive and abundant substrate, 2-phenylethanol. We have recently shown that combining random mutagenesis, saturation mutagenesis, and statistical modeling, enables the creation of highly active enzyme variants. Thus, variant TmoA I100A/E214G/D285Q of T4MO was 200-fold more active on 2-phenylethanol than wild-type.
This project is funded by the Technion-Haifa University Collaboration Fund
Evolving T4MO for PEA hydroxylation by the aid of a statistical model