| Title : High-throughput selection of (new) enzymes: phage display-mediated isolation of alkyl halide hydrolases from a library of active-site mutated epoxide hydrolases - Blazic_2024_Faraday.Discuss__ |
| Author(s) : Blazic M , Gautier C , Norberg T , Widersten M |
| Ref : Faraday Discussion , : , 2024 |
|
Abstract :
Epoxide hydrolase StEH1, from potato, is similar in overall structural fold and catalytic mechanism to haloalkane dehalogenase DhlA from Xanthobacter autotrophicus. StEH1 displays low (promiscuous) hydrolytic activity with (2-chloro)- and (2-bromo)ethanebenzene producing 2-phenylethanol. To investigate possibilities to amplify these very low dehalogenase activities, StEH1 was subjected to targeted randomized mutagenesis at five active-site amino acid residues and the resulting protein library was challenged for reactivity towards a bait chloride substrate. Enzymes catalyzing the first half-reaction of a hydrolytic cycle were isolated following monovalent phage display of the mutated proteins. Several StEH1 derived enzymes were identified with enhanced dehalogenase activities. |
| PubMedSearch : Blazic_2024_Faraday.Discuss__ |
| PubMedID: 38828992 |
| Gene_locus related to this paper: soltu-3epoxy |
| Inhibitor | Biotin-PEG3-(2-Chloroethyl)benzene |
| Substrate | (2-Bromoethyl)benzene |
| Gene_locus | soltu-3epoxy |
Blazic M, Gautier C, Norberg T, Widersten M (2024)
High-throughput selection of (new) enzymes: phage display-mediated isolation of alkyl halide hydrolases from a library of active-site mutated epoxide hydrolases
Faraday Discussion
:
Blazic M, Gautier C, Norberg T, Widersten M (2024)
Faraday Discussion
: