Synergism with novel expansin-related proteins for cellulose processing

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School of Chemical Engineering | Doctoral thesis (article-based) | Defence date: 2024-12-17
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Authors

Date

2024

Department

Biotuotteiden ja biotekniikan laitos
Department of Bioproducts and Biosystems

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Mcode

Degree programme

Language

en

Pages

75 + app. 133

Series

Aalto University publication series DOCTORAL THESES, 266/2024

Abstract

The demand for environmentally sustainable materials has accelerated the use of renewable biomass, such as lignocellulose from plant fibers. This study focuses on innovative biotechnological methods to enhance the conversion of lignocellulosic biomass into valuable products, thereby promoting circular economy principles. Specifically, the research investigates the production and application of microbial expansin-related proteins (EXLX), particularly loosenin-like proteins, to improve enzymatic hydrolysis processes for biomass conversion.  The study employs Pichia pastoris strains SMD1168H and KM71H to produce and scale up these proteins, and further assessing their impact on converting softwood kraft pulp to nanocellulose. Key objectives include evaluating these proteins' ability to synergize with cellulolytic enzymes, enhancing sugar release from softwood kraft pulps, and facilitating the production of cellulose nanocrystals (CNCs). Advanced techniques such as Biological Small Angle Neutron Scattering (Bio-SANS) were used to analyze the structural impact of the proteins on holocellulose.  Results demonstrate that recombinant expansin-related proteins can significantly boost enzymatic hydrolysis efficiency. This enhancement is attributed to the proteins' ability to increase enzyme accessibility by modifying fiber morphology. The integration of these proteins in enzymatic cocktails showed a marked improvement in glucose and xylose yields from various cellulosic substrates. Furthermore, the study explores the enzymatic production of CNCs with enhanced stability and dispersibility, suitable for applications in conductive inks and other advanced materials. The findings underscore the potential of these proteins to reduce enzyme loadings, thereby lowering production costs and environmental impact. Future research directions include optimizing process parameters for large-scale applications and investigating synergistic effects with other enzyme systems.  In conclusion, the incorporation of microbial expansin-related proteins in biomass processing holds significant promise for advancing sustainable production technologies. This research provides foundational insights into their mode of action and range of applications, paving the way for more efficient and eco-friendly biomass conversion methods.

Description

Supervising professor

Master, Emma, Prof., Aalto University, Department of Bioproducts and Biosystems, Finland

Thesis advisor

Maloney, Thaddeus, Prof., Aalto University, Department of Bioproducts and Biosystems, Finland

Keywords

biomass conversion, bioreactors, lignocellulose, enzymatic hydrolysis, fibrillation, fiber structure changes, Loosenin-like proteins from Phanerochaete carnosa (PcaLOOLs), protein production, sugar release, synergistic enzymatic treatments

Other note

Parts

  • [Publication 1]: Deepika Dahiya, Sara Ceccherini, and Thad C. Maloney. “Impact of high consistency enzymatic hydrolysis and defibration drying on cellulose fiber pore characteristics.” Cellulose 30.12 (2023): 7607-7618.
    Full text in Acris/Aaltodoc: https://urn.fi/URN:NBN:fi:aalto-202308305350
    DOI: 10.1007/s10570-023-05398-z View at publisher
  • [Publication 2]: Deepika Dahiya, Taru Koitto, Kim Kutvonen, Yan Wang, Majid Haddad Momeni, Siiri de Ruijter, and Emma R. Master. "Fungal loosenin-like proteins boost the cellulolytic enzyme conversion of pretreated wood fiber and cellulosic pulps." Bioresource Technology 394 (2024): 130188.
    Full text in Acris/Aaltodoc: https://urn.fi/URN:NBN:fi:aalto-202401171473
    DOI: 10.1016/j.biortech.2023.130188 View at publisher
  • [Publication 3]: Ylenia Jabalera*, Deepika Dahiya*, Carlos David Ordóñez Cencerrado, Antonio Jesus Caballero, Nerea Zaldua, Arantxa Eceiza, Emma R. Master, and Raul Perez-Jimenez. " Impact of loosenins on the enzymatic preparation of cellulose nanocrystals.” (Under review in Carbohydrate Polymers) *Co-first authors
  • [Publication 4]: Deepika Dahiya, Zsuzsanna Péter-Szabó, Manjula Senanayake, Sai Venkatesh Pingali, Wellington C. Leite, James Byrnes, Garry W. Buchko, Pramod Sivan, Francisco Vilaplana, Emma R. Master and Hugh O’Neill. "SANS investigation of fungal loosenins reveal substrate dependent impacts of protein action on inter-fibril distance and packing order of cellulosic substrate.” (Under review in Biotechnology for Biofuels and Bioproducts).
    DOI: 10.21203/rs.3.rs-4769386/v1 View at publisher

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https://urn.fi/URN:ISBN:978-952-64-2174-2

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[diss] Kemian tekniikan korkeakoulu / CHEM