The monolignol oxidoreductase AtBBE-like protein 15: Characterization, rational engineering and biocatalytic applications.

Bastian Daniel, Julia Messenlehner, Jörg H Schrittwieser, Sabine Pils, Wolfgang Kroutil, Peter Macheroux

Research output: Contribution to conferencePoster

Abstract

Flavoproteins are a diverse protein class employing an isoalloxazine ring for catalysis in form of the flavin mononucleotide (FMN) or the flavin adenine dinucleotide (FAD). Among them is the berberine bridge enzyme-like (BBE-like) protein family 1. Monolignol oxidoreductases from this protein family catalyze the oxidation of monolignols to the corresponding aldehydes 2. Recently we explored the potential of a monolignol oxidoreductase from Arabidopsis thaliana (AtBBE-like protein 15) as bio-catalyst 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)3. 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 min. The enzyme shows increased activity in the presence of various co-solvents (10–50% v/v. Primary benzylic and primary or secondary allylic alcohols were accepted as substrates. The enantioselectivity in the oxidation of secondary alcohols was good to excellent (E>34 to >200). As the enzyme was found to be a versatile and robust biocatalyst we expanded our mutagenesis program to elucidate the catalytic mechanism (presented at the same conference by Julia Messenlehner), to broaden the substrate scope and to alter the enantioselectivity. 1.Daniel, B. et al. The family of berberine bridge enzyme-like enzymes: A treasure-trove of oxidative reactions. Arch. Biochem. Biophys. 632, (2017).2.Daniel, B. et al. Oxidation of Monolignols by Members of the Berberine Bridge Enzyme Family Suggests a Role in Cell Wall Metabolism. J. Biol. Chem. 290, 18770–18781 (2015).3.Pils, S. et al. Characterization of the monolignol oxidoreductase AtBBE-like protein 15 L182V for biocatalytic applications. J. Mol. Catal. B Enzym. (2016). doi:10.1016/j.molcatb.2016.10.018
Original languageEnglish
Publication statusPublished - 28 Aug 2018
Event9th International Congress on Biocatalysis: 9th International Congress on Biocatalysis - Hamburg University of Technology TUHH, Hamburg, Germany
Duration: 26 Aug 201830 Aug 2018

Conference

Conference9th International Congress on Biocatalysis
Abbreviated titleBiocat 2018
CountryGermany
CityHamburg
Period26/08/1830/08/18

ASJC Scopus subject areas

  • Biochemistry
  • Catalysis
  • Biotechnology
  • Bioengineering

Fields of Expertise

  • Human- & Biotechnology

Treatment code (Nähere Zuordnung)

  • Application

Cite this

Daniel, B., Messenlehner, J., Schrittwieser, J. H., Pils, S., Kroutil, W., & Macheroux, P. (2018). The monolignol oxidoreductase AtBBE-like protein 15: Characterization, rational engineering and biocatalytic applications.. Poster session presented at 9th International Congress on Biocatalysis, Hamburg, Germany.