Busk, Peter Kamp

Grell, Morten Nedergaard

Lange, Lene

Lange, Mette

Discovery of novel GH61 and attempt to correlate subfamily grouping with taxonomy and physiology of producing organismMette Lange, Peter Kamp Busk, Morten Nedergaard Grell and Lene LangeAalborg University Copenhagen, AAU Cph, Department of Biotechnology, Chemistry and Environmental Engineering. mla@bio.aau.dkIn the continuous search, for even more effective enzymes for biomass degradation the glycoside hydrolase (GH) family 61 has been shown to be an important auxiliary protein in fungal degradation of lignocellulosic biomass. There are 152 fungal entries for GH61 in the CAZY database and so far no subgroupings have been identified. A conserved domain search in the protein databases available at NCBI (http://NCBI.nlm.nih.gov.protein/) identifies more than 467 proteins from 51 different fungi, thus a single species can have the genetic capacity to produce several GH61 proteins. With the aim of understanding the biology behind the various genetic capacities of the GH61 genes in each fungus, we are analysing the gene expression pattern of the set of GH61 genes present in a single fungus, by challenging the fungus with different substrates and analysing the gene expression pattern by quantitative real time PCR. We are also aiming at discovering new GH61 proteins from relevant biomass degrading fungi. Due to low sequence similarity a conventional degenerated primer approach based on sequence alignments has low success rate, thus the new tool for Peptide Pattern Recognition (PPR) was applied (Busk PK and Lange L, AAU patent). PPR divided the GH61 family into a number of comprehensive groups and identified the conserved motifs in each group and showed that one group of GH61 proteins was specifically present among the basidiomycetes. This allowed us to target the degenerated primer design for this specific subgroup and resulted in successfully identification of novel GH61 sequences from 5 basidiomycetous fungi. At present we are working on the heterologous expression and purification of the identified GH61 proteins.

Discovery of novel GH61 and attempt to correlate subfamily grouping with taxonomy and physiology of producing organism

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