Characterization of the monolignol oxidoreductase AtBBE-like protein 15 L182V for biocatalytic applications

Sabine Pils, Kordula Schnabl, Silvia Wallner, Marko Kljajic, Nina Kupresanin, Rolf Breinbauer, Jörg Schrittwieser, Michael Fuchs, Wolfgang Kroutil, Bastian Daniel, Peter Macheroux

Research output: Contribution to journalArticle

Abstract

Monolignol oxidoreductases from the berberine bridge enzyme-like (BBE-like) protein family (pfam 08031) catalyze the oxidation of monolignols to the corresponding aldehydes. In this report, we explore the potential of a monolignol oxidoreductase from Arabidopsis thaliana (AtBBE-like protein 15) as biocatalyst for oxidative reactions. For this study we employed a variant with enhanced reactivity towards oxygen, which was obtained by a single amino acid exchange (L182V). The pH and temperature optima of the purified AtBBE-like protein 15 L182V were determined as well as the tolerance toward organic co-solvents; furthermore the substrate scope was characterized. The enzyme has a temperature optimum of 50 °C and retains more than 50% activity between pH 5 and pH 10 within 5 minutes. The enzyme shows increased activity in the presence of various co-solvents (10-50% v/v), including acetonitrile, 2-propanol, 1,4-dioxane, and dimethyl sulfoxide. Primary benzylic and primary or secondary allylic alcohols were accepted as substrates. The enantioselectivity E in the oxidation of secondary alcohols was good to excellent (E >34 to >200).
Original languageEnglish
Pages (from-to)S6-S14
JournalJournal of molecular catalysis / B
Volume133
Issue numberSuppl.1
DOIs
Publication statusPublished - 2016

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