Loosenin-Like Proteins from Phanerochaete carnosa Impact Both Cellulose and Chitin Fiber Networks

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Journal Title
Journal ISSN
Volume Title
A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
Date
2023-01
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Mcode
Degree programme
Language
en
Pages
15
Series
Applied and Environmental Microbiology, Volume 89, issue 1
Abstract
Microbial expansin-related proteins are ubiquitous across bacterial and fungal organisms and reportedly play a role in the modification and deconstruction of cell wall polysaccharides, including lignocellulose. So far, very few microbial expansin-related proteins, including loosenins and loosenin-like (LOOL) proteins, have been functionally characterized. Herein, four LOOLs encoded by Phanerochaete carnosa and belonging to different subfamilies (i.e., PcaLOOL7 and PcaLOOL9 from subfamily A and PcaLOOL2 and PcaLOOL12 from subfamily B) were recombinantly produced and the purified proteins were characterized using diverse cellulose and chitin substrates. The purified PcaLOOLs weakened cellulose filter paper and cellulose nanofibril networks (CNF); however, none significantly boosted cellulase activity on the selected cellulose substrates (Avicel and Whatman paper). Although fusing the family 63 carbohydrate-binding module (CBM63) of BsEXLX1 encoded by Bacillus subtilis to PcaLOOLs increased their binding to cellulose, the CBM63 fusion appeared to reduce the cellulose filter paper weakening observed using wild-type proteins. Binding of PcaLOOLs to alpha-chitin was considerably higher than that to cellulose (Avicel) and was pH dependent, with the highest binding at pH 5.0. Amendment of certain PcaLOOLs in fungal liquid cultivations also impacted the density of the cultivated mycelia. The present study reveals the potential of fungal expansin-related proteins to impact both cellulose and chitin networks and points to a possible biological role in fungal cell wall processing.
Description
| openaire: EC/H2020/648925/EU//BHIVE | openaire: EC/H2020/964764/EU//BioUPGRADE Funding Information: We thank Thu Vuong (Master Group, University of Toronto) for his assistance with generating model protein structures. We also acknowledge the use of Aalto University Bioeconomy facilities as well as the Biocenter Oulu Protein Biophysical Analysis core facility (a member of Biocenter Finland). We declare that no competing interests exist. This project received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no. 964764 and the European Research Council (ERC) Consolidator program (grant no. BHIVE–648925). This study was also supported by funding from the Novo Nordisk Foundation (BIOSEMBL–34622). Work by J.V. and K.S.M. was supported by the Academy of Finland under grant no. 311244; work by D.J.C. and E.R.W. was supported by the U.S. Department of Energy, Office of Science, Basic Energy Sciences, under grant no. DE-FG02-84ER13179, and work by T.K. was supported by the Jenny and Antti Wihuri Foundation (Centre for Young Synbio Scientists). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The content presented in this article represents the views of the authors, and the European Commission is not responsible for any use that may be made of the information it contains. Funding Information: This project received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no. 964764 and the European Research Council (ERC) Consolidator program (grant no. BHIVE–648925). This study was also supported by funding from the Novo Nordisk Foundation (BIOSEMBL–34622). Work by J.V. and K.S.M. was supported by the Academy of Finland under grant no. 311244; work by D.J.C. and E.R.W. was supported by the U.S. Department of Energy, Office of Science, Basic Energy Sciences, under grant no. DE-FG02-84ER13179, and work by T.K. was supported by the Jenny and Antti Wihuri Foundation (Centre for Young Synbio Scientists). Publisher Copyright: Copyright © 2023 Monschein et al.
Keywords
biotechnology, carbohydrate active enzymes, carbohydrate binding, cellulose, chitin, expansin, filamentous fungi, lignocellulose, loosenin, Phanerochaete carnosa
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Citation
Monschein , M , Ioannou , E , Koitto , T , AI Amin , L A K M , Varis , J J , Wagner , E R , Mikkonen , K S , Cosgrove , D J & Master , E R 2023 , ' Loosenin-Like Proteins from Phanerochaete carnosa Impact Both Cellulose and Chitin Fiber Networks ' , Applied and Environmental Microbiology , vol. 89 , no. 1 . https://doi.org/10.1128/aem.01863-22