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

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

doi:10.1186/s13068-024-02500-w

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

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

Research output: Contribution to journal › Article › Scientific › peer-review

Abstract

Background: 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. Results: 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%. Conclusion: 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.

Original language	English
Article number	56
Journal	Biotechnology for Biofuels and Bioproducts
Volume	17
Issue number	1
DOIs	https://doi.org/10.1186/s13068-024-02500-w
Publication status	Published - Dec 2024
MoE publication type	A1 Journal article-refereed

Funding

This project has received funding from the European Union\u2019s Horizon 2020 research and innovation program under grant agreement No 964764. The project also received funding from the European Union\u2014Next Generation EU instrument and the Academy of Finland under grant number 347612 (MIMIC). This project was further supported by the Academy of Finland (Grant no. 338804 and 347612) and Business Finland. AZ was supported through the FinnCERES Materials Bioeconomy Ecosystem. A portion of this research was performed on a project award (60027) under the FICUS program, which provided access to resources at the Joint Genome Institute and the Environmental Molecular Sciences Laboratory, which are DOE Office of Science User Facilities. Both facilities are sponsored by the Biological and Environmental Research program and operated under Contract Nos. DE-AC02-05CH11231 (JGI) and DE-AC05-76RL01830 (EMSL).

Keywords

Carbohydrate-active enzymes
Cellulose
Defibrillation
Expansin
Non-lytic

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Haddad Momeni, M., Zitting, A., Jäämuru, V., Turunen, R., Penttilä, P., Buchko, G. W., Hiltunen, S., Maiorova, N., Koivula, A., Sapkota, J., Marjamaa, K., & Master, E. R. (2024). Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils. Biotechnology for Biofuels and Bioproducts, 17(1), Article 56. https://doi.org/10.1186/s13068-024-02500-w

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title = "Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils",

abstract = "Background: 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. Results: 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%. Conclusion: 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.",

keywords = "Carbohydrate-active enzymes, Cellulose, Defibrillation, Expansin, Non-lytic",

author = "{Haddad Momeni}, Majid and Aleksi Zitting and Vilma J{\"a}{\"a}muru and Rosaliina Turunen and Paavo Penttil{\"a} and Buchko, {Garry W.} and Salla Hiltunen and Natalia Maiorova and Anu Koivula and Janak Sapkota and Kaisa Marjamaa and Master, {Emma R.}",

year = "2024",

month = dec,

doi = "10.1186/s13068-024-02500-w",

language = "English",

volume = "17",

journal = "Biotechnology for Biofuels and Bioproducts",

issn = "2731-3654",

publisher = "BioMed Central",

number = "1",

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Haddad Momeni, M, Zitting, A, Jäämuru, V, Turunen, R, Penttilä, P, Buchko, GW, Hiltunen, S, Maiorova, N , Koivula, A, Sapkota, J, Marjamaa, K & Master, ER 2024, 'Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils', Biotechnology for Biofuels and Bioproducts, vol. 17, no. 1, 56. https://doi.org/10.1186/s13068-024-02500-w

TY - JOUR

T1 - Insights into the action of phylogenetically diverse microbial expansins on the structure of cellulose microfibrils

AU - Haddad Momeni, Majid

AU - Zitting, Aleksi

AU - Jäämuru, Vilma

AU - Turunen, Rosaliina

AU - Penttilä, Paavo

AU - Buchko, Garry W.

AU - Hiltunen, Salla

AU - Maiorova, Natalia

AU - Koivula, Anu

AU - Sapkota, Janak

AU - Marjamaa, Kaisa

AU - Master, Emma R.

PY - 2024/12

Y1 - 2024/12

N2 - Background: 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. Results: 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%. Conclusion: 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.

AB - Background: 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. Results: 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%. Conclusion: 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.

KW - Carbohydrate-active enzymes

KW - Cellulose

KW - Defibrillation

KW - Expansin

KW - Non-lytic

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U2 - 10.1186/s13068-024-02500-w

DO - 10.1186/s13068-024-02500-w

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AN - SCOPUS:85191092236

SN - 2731-3654

VL - 17

JO - Biotechnology for Biofuels and Bioproducts

JF - Biotechnology for Biofuels and Bioproducts

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M1 - 56

ER -

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

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