Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils

Momeni, Majid Haddad; Zitting, Aleksi; Jäämuru, Vilma; Turunen, Rosaliina; Penttilä, Paavo; Buchko, Garry W.; Hiltunen, Salla; Maiorova, Natalia; Koivula, Anu; Sapkota, Janak; Marjamaa, Kaisa; Master, Emma R.

doi:10.1186/s13068-024-02500-w

Title: Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils

Journal Article · 22 April 2024 · Biotechnology for Biofuels and Bioproducts

DOI: https://doi.org/10.1186/s13068-024-02500-w · OSTI ID:2341731

Momeni, Majid Haddad ^[1]; Zitting, Aleksi ^[1]; Jäämuru, Vilma ^[1]; Turunen, Rosaliina ^[1]; Penttilä, Paavo ^[1];

^[2]; Hiltunen, Salla ^[3]; Maiorova, Natalia ^[4]; Koivula, Anu ^[4]; Sapkota, Janak ^[3]; Marjamaa, Kaisa ^[4]; Master, Emma R. ^[5]

Aalto University, Otaniemi (Finland)
Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Washington State University, Pullman, WA (United States)
NE Research Center, UPM Pulp Research and Innovations, Lappeenranta (Finland)
VTT Technical Research Centre of Finland Ltd., Espoo (Finland)
Aalto University, Otaniemi (Finland); University of Toronto, ON (Canada)

Microbial expansins (EXLXs) are non-lytic proteins homologous to plant expansins involved in plant cell wall formation. Due to their non-lytic cell wall loosening properties and potential to disaggregate cellulosic structures, there is considerable interest in exploring the ability of microbial expansins (EXLX) to assist the processing of cellulosic biomass for broader biotechnological applications. Herein, EXLXs with different modular structure and from diverse phylogenetic origin were compared in terms of ability to bind cellulosic, xylosic, and chitinous substrates, to structurally modify cellulosic fibrils, and to boost enzymatic deconstruction of hardwood pulp. Five heterogeneously produced EXLXs (Clavibacter michiganensis; CmiEXLX2, Dickeya aquatica; DaqEXLX1, Xanthomonas sacchari; XsaEXLX1, Nothophytophthora sp.; NspEXLX1 and Phytophthora cactorum; PcaEXLX1) were shown to bind xylan and hardwood pulp at pH 5.5 and CmiEXLX2 (harboring a family-2 carbohydrate-binding module) also bound well to crystalline cellulose. Small-angle X-ray scattering revealed a 20–25% increase in interfibrillar distance between neighboring cellulose microfibrils following treatment with CmiEXLX2, DaqEXLX1, or NspEXLX1. Correspondingly, combining xylanase with CmiEXLX2 and DaqEXLX1 increased product yield from hardwood pulp by ~ 25%, while supplementing the TrAA9A LPMO from Trichoderma reesei with CmiEXLX2, DaqEXLX1, and NspEXLX1 increased total product yield by over 35%. This direct comparison of diverse EXLXs revealed consistent impacts on interfibrillar spacing of cellulose microfibers and performance of carbohydrate-active enzymes predicted to act on fiber surfaces. These findings uncover new possibilities to employ EXLXs in the creation of value-added materials from cellulosic biomass.

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Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

Sponsoring Organization:: Academy of Finland; European Union’s Horizon 2020; FinnCERES Materials Bioeconomy Ecosystem; USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231; AC05-76RL01830

OSTI ID:: 2341731

Report Number(s):: PNNL-SA-187033

Journal Information:: Biotechnology for Biofuels and Bioproducts, Journal Name: Biotechnology for Biofuels and Bioproducts Journal Issue: 1 Vol. 17; ISSN 2731-3654

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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Title: Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils

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