Discovery of catalytic phages by biocatalytic self-assembly

Maeda, Yoshiaki and Javid, Nadeem and Duncan, Krystyna and Birchall, Louise and Gibson, Kirsty F and Cannon, Daniel and Kanetsuki, Yuka and Knapp, Charles W and Tuttle, Tell and Ulijn, Rein V. and Matsui, Hiroshi (2014) Discovery of catalytic phages by biocatalytic self-assembly. Journal of the American Chemical Society, 136 (45). 15893–15896. ISSN 0002-7863

Preview

PDF (Ulijn-JACS-discovery-of-catalytic-phages-23-October-2014) Ulijn_JACS_discovery_of_catalytic_phages_23_October_2014.pdf Accepted Author Manuscript Download (833kB)| Preview

Abstract

Discovery of new catalysts for demanding aqueous reactions is challenging. In here, we describe methodology for selection of catalytic phages by taking advantage of localized assembly of the product of the catalytic reaction that is screened for. A phage display library covering 109 unique dodecapeptide sequences is incubated with non-assembling precursors. Phages which are able to catalyze formation of the self-assembling reaction product (via amide condensation) acquire an aggregate of reaction product, enabling separation by centrifugation. The thus selected phages can be ampli-fied by infection of E.coli. These phages are shown to catalyze amide condensation and hydrolysis. Kinetic analysis shows a minor role for substrate binding. The approach enables discovery and mass-production of biocatalytic phages.