A family AA5_2 carbohydrate oxidase from Penicillium rubens displays functional overlap across the AA5 family

Filip Mollerup; Ville Aumala; Kirsti Parikka; Yann Mathieu; Harry Brumer; Maija Tenkanen; Emma Master

doi:10.1371/journal.pone.0216546

A family AA5_2 carbohydrate oxidase from Penicillium rubens displays functional overlap across the AA5 family

PLoS One. 2019 May 15;14(5):e0216546. doi: 10.1371/journal.pone.0216546. eCollection 2019.

Authors

Filip Mollerup¹, Ville Aumala¹, Kirsti Parikka², Yann Mathieu³, Harry Brumer³, Maija Tenkanen², Emma Master^{1

4}

Affiliations

¹ Department of Bioproducts and Biosystems, Aalto University, Aalto, Finland.
² Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.
³ Michael Smith Laboratories, University of British Columbia, Vancouver, British Columbia, Canada.
⁴ Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.

Abstract

Copper radical alcohol oxidases belonging to auxiliary activity family 5, subfamily 2 (AA5_2) catalyze the oxidation of galactose and galactosides, as well as aliphatic alcohols. Despite their broad applied potential, so far very few AA5_2 members have been biochemically characterized. We report the recombinant production and biochemical characterization of an AA5_2 oxidase from Penicillium rubens Wisconsin 54-1255 (PruAA5_2A), which groups within an unmapped clade phylogenetically distant from those comprising AA5_2 members characterized to date. PruAA5_2 preferentially oxidized raffinose over galactose; however, its catalytic efficiency was 6.5 times higher on glycolaldehyde dimer compared to raffinose. Deep sequence analysis of characterized AA5_2 members highlighted amino acid pairs correlated to substrate range and conserved within the family. Moreover, PruAA5_2 activity spans substrate preferences previously reported for AA5 subfamily 1 and 2 members, identifying possible functional overlap across the AA5 family.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cloning, Molecular / methods*
Fungal Proteins / genetics
Fungal Proteins / metabolism
Galactose / chemistry
Galactosides / chemistry
High-Throughput Nucleotide Sequencing
Oxidation-Reduction
Oxidoreductases / genetics*
Oxidoreductases / metabolism*
Penicillium / enzymology*
Penicillium / genetics
Phylogeny
Protein Engineering
Raffinose / chemistry*
Recombinant Proteins / metabolism
Sequence Analysis, Protein

Substances

Fungal Proteins
Galactosides
Recombinant Proteins
Oxidoreductases
Raffinose
Galactose

Grants and funding

This study was financially supported by the European Research Council (ERC) (Consolidator Grant no. BHIVE – 648925) (FM, VA and EM); Academy of Finland (252183)(KP and MT), an NSERC Discovery Grant and NSERC Strategic Partnership Grant as well as the Genome Canada funded Synbiomics project (10405) (YM and HB) with support from Ontario Genomics, Genome Quebec, and Genome BC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.