Fungal glucuronoyl esterases: Genome mining based enzyme discovery and biochemical characterization
Adiphol Dilokpimol, Miia R. Mäkelä, Gabriella Cerullo, Miaomiao Zhou, Simona Varrial, Loknath Gidijal, Joana L.A. Brás, Peter Jütten, Alexander Piechot, Raymond Verhaert, Vincenza Faraco, Kristiina S. Hilden, Ronald P. de Vries
New Biotechnol. 40, Jan. 2018, pp 282-287 DOI:10.1016/j.nbt.2017.10.003;
4-O-Methyl-d-glucuronic acid (MeGlcA) is a side-residue of glucuronoarabinoxylan and can form ester linkages to lignin, contributing significantly to the strength and rigidity of the plant cell wall. Glucuronoyl esterases (4-O-methyl-glucuronoyl methylesterases, GEs) can cleave this ester bond, and therefore may play a significant role as auxiliary enzymes in biomass saccharification for the production of biofuels and biochemicals. GEs belong to a relatively new family of carbohydrate esterases (CE15) in the CAZy database (www.cazy.org), and so far around ten fungal GEs have been characterized. To explore additional GE enzymes, we used a genome mining strategy. BLAST analysis with characterized GEs against approximately 250 publicly accessible fungal genomes identified more than 150 putative fungal GEs, which were classified into eight phylogenetic sub-groups. To validate the genome mining strategy, 21 selected GEs from both ascomycete and basidiomycete fungi were heterologously produced in Pichia pastoris. Of these enzymes, 18 were active against benzyl d-glucuronate demonstrating the suitability of our genome mining strategy for enzyme discovery.
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