Publication

Directed Evolution of a Designer Enzyme Featuring an Unnatural Catalytic Amino Acid

Mayer, C., Dulson, C., Reddem, E., Thunnissen, A-M. W. H. & Roelfes, G., 11-Feb-2019, In : Angewandte Chemie-International Edition. 58, 7, p. 2083-2087 5 p.

Research output: Contribution to journalArticleAcademicpeer-review

The impressive rate accelerations that enzymes display in nature often result from boosting the inherent catalytic activities of side chains by their precise positioning inside a protein binding pocket. Such fine-tuning is also possible for catalytic unnatural amino acids. Specifically, the directed evolution of a recently described designer enzyme, which utilizes an aniline side chain to promote a model hydrazone formation reaction, is reported. Consecutive rounds of directed evolution identified several mutations in the promiscuous binding pocket, in which the unnatural amino acid is embedded in the starting catalyst. When combined, these mutations boost the turnover frequency (k(cat)) of the designer enzyme by almost 100-fold. This results from strengthening the catalytic contribution of the unnatural amino acid, as the engineered designer enzymes outperform variants, in which the aniline side chain is replaced with a catalytically inactive tyrosine residue, by more than 200-fold.

Original languageEnglish
Pages (from-to)2083-2087
Number of pages5
JournalAngewandte Chemie-International Edition
Volume58
Issue number7
Publication statusPublished - 11-Feb-2019

    Keywords

  • directed evolution, enzyme catalysis, enzyme design, hydrazones, organocatalysis, NUCLEOPHILIC CATALYSIS, HYDRAZONE, BIOCATALYSIS, MECHANISM, UNIVERSE

View graph of relations

Download statistics

No data available

ID: 77009913