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Deciphering lignin heterogeneity in ball milled softwood: unravelling the synergy between the supramolecular cell wall structure and molecular events
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Glycoscience. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Centres, Wallenberg Wood Science Center.ORCID iD: 0000-0002-7738-5952
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Fibre- and Polymer Technology, Wood Chemistry and Pulp Technology. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Centres, Wallenberg Wood Science Center.ORCID iD: 0000-0002-8614-6291
2021 (English)In: Green Chemistry, ISSN 1463-9262, E-ISSN 1463-9270, Vol. 23, no 9, p. 3348-3364Article in journal (Refereed) Published
Abstract [en]

Mechanical milling of lignocellulose has been used in several studies as a key pretreatment enabling the extraction of lignin from various sources for structural analysis. It is also applied as an alternative to wet chemical methods for lignin valorization. However, the changes caused to the plant cell walls at different hierarchical scales and how they relate to the molecular events are still poorly understood. In this context, we sought to gain deeper insights into molecular heterogeneity in milled cell walls, with a primary focus on lignin. A novel fractionation protocol was developed to enable the advanced analysis (1D and 2D NMR, SEC, XRD) of molecular populations in ball milled fiber walls. The methodology was applied to follow the emergence of such populations through the milling process, and in different milling environments. Lignin heterogeneity in the ball milled fibers was found to consist of distinct populations of small and large fractions of lignin carbohydrate complexes and pure lignin fractions, both with differences in lignin inter-unit abundances. Lignin-carbohydrate bonds of benzyl ester type were unequivocally demonstrated for the first time by combination of HSQC-HMBC NMR analysis. γ-Ester LCC and phenyl glycoside LCCs were also detected. Furthermore, an important branching point in lignin, previously controversial, namely the 4-O-etherified 5-5′ substructure, is unequivocally shown here by HSQC-HMBC analysis of the milled wood isolates, and supported by biomimetic lignin (DHP) to originate from the native structure. Based on the advanced characterization, the origin of lignin heterogeneity in ball milled fibers is proposed to result from the uneven distribution of the applied mechanical energy, where synergistic effects between crystalline and amorphous states play a central role. Accordingly, a plant cell wall model is proposed and a complete mechanism of its disintegration during the milling exercise is presented. The unveiled heterogeneity model of ball milled cell walls can serve as a useful guide for future studies on mechanical fractionation and valorization of lignocellulose based polymers.

Place, publisher, year, edition, pages
Royal Society of Chemistry (RSC) , 2021. Vol. 23, no 9, p. 3348-3364
National Category
Wood Science
Identifiers
URN: urn:nbn:se:kth:diva-309868DOI: 10.1039/d0gc04319bISI: 000637012300001Scopus ID: 2-s2.0-85105851035OAI: oai:DiVA.org:kth-309868DiVA, id: diva2:1644405
Funder
Knut and Alice Wallenberg Foundation
Note

QC 20220314

Available from: 2022-03-14 Created: 2022-03-14 Last updated: 2023-05-12Bibliographically approved
In thesis
1. Biosynthesis, interactions, and structure of native lignin
Open this publication in new window or tab >>Biosynthesis, interactions, and structure of native lignin
2023 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Wood biomass is an important resource in the development ofstrategies towards replacement of fossil-based materials. Woodcomprises cellulose, hemicelluloses and lignin, in amounts thatvary between species. Traditionally, the extraction of cellulose hasbeen prioritized for the production of pulp, paper, and novelmaterials, at the expense of the other two thirds of the woodbiomass. Lignin is an important component of wood, comprising acomplex polymer of aromatic compounds. At the moment, mostof this abundant biopolymer is usually burned for energy or simplydiscarded as a huge waste of the pulp industries, after severemodifications in its already complex structure have rendered itdifficult to valorize. A deeper understanding of the lignin polymerand its properties can guide the design of milder extractiontechniques to obtain specific lignin structures as are needed.In this thesis, fundamental aspects of lignin biosynthesis,structure, and interactions with other cell wall components areinvestigated, employing model biological systems and diversechemical methods. Specifically, a Norway spruce tissue culturethat produces lignin extracellularly, without deposition to theplant cell wall, was used to understand the impact of a secondarycell wall hemicellulose on the production and structure of lignin.The main advantage of this model system is that it enables thecollection of lignin without the need for extraction, which is knownto alter the structure of lignin. The same system was studied viatranscriptomic analysis to elucidate the impact of a hemicelluloseon the lignin biosynthetic pathway and other metabolic processes.The effect of extraction on the structure of lignin was addressedwith the development of mild and green extraction protocols forsoftwood and hardwood species. Different conditions led to thecollection of discrete lignin populations. The results could informthe development of lignin-first biorefinery applications, in whichspecific structural properties of the biopolymer are preserved.Finally, native-like lignin fractions extracted using these mildprotocols were used for the preparation of lignin nanoparticles, toshowcase their potential in high-end applications. Furthermore,the importance of unmodified lignin in the properties of woodaerogel applications was also studied, and was included in thisthesis to demonstrate that lignin modification or even removal isnot always necessary to obtain materials with propertiescompetitive to their fossil-based analogues.

Abstract [sv]

Träbiomassa är en viktig resurs i utvecklingen av strategier för attersätta fossilbaserade material. Trä består av cellulosa,hemicellulosa och lignin, i mängder som varierar mellan arterna.Traditionellt har utvinning av cellulosa prioriterats förframställning av massa, papper och nya material, på bekostnad avövriga två tredjedelar av träbiomassan. Lignin är en viktigkomponent i trä, som består av en komplex polymer av aromatiskaföreningar. För närvarande bränns det mesta av denna rikligabiopolymer vanligtvis för energi eller helt enkelt kasseras som ettenormt slöseri från massaindustrin, efter att allvarligamodifieringar i dess redan komplexa struktur har gjort det svårt attvärdera den. En djupare förståelse av ligninpolymeren och dessegenskaper kan vägleda utformningen av mildareextraktionstekniker för att erhålla specifika ligninstrukturer efterbehov.I denna avhandling undersöks grundläggande aspekter avligninbiosyntes, struktur och interaktioner med andracellväggskomponenter, med hjälp av biologiska modellsystem ocholika kemiska metoder. Specifikt användes en granvävnadskultursom producerar lignin extracellulärt, utan avsättning påväxtcellväggen, för att förstå effekten av en sekundärcellväggshemicellulosa på produktionen och strukturen av lignin.Den största fördelen med detta modellsystem är att det möjliggörinsamling av lignin utan behov av extraktion, vilket är känt för attförändra strukturen av lignin. Samma system studerades viatranskriptomisk analys för att belysa inverkan av en hemicellulosapå ligninbiosyntesvägen och andra metaboliska processer.Effekten av extraktion på strukturen av lignin togs upp medutvecklingen av milda och gröna extraktionsprotokoll för barr- ochlövvedsarter. Olika förhållanden ledde till insamling av specifikaligninpopulationer. Resultaten skulle kunna informera omutvecklingen av lignin-första bioraffinaderiapplikationer, därspecifika strukturella egenskaper hos biopolymeren bevaras.Slutligen användes naturliga ligninfraktioner extraherade meddessa milda protokoll för framställning av ligninnanopartiklar föratt visa upp deras potential i avancerade applikationer. Vidarestuderades också betydelsen av omodifierat lignin i egenskapernahos aerogelapplikationer i trä, och inkluderades i dennaavhandling för att visa att modifiering eller till och med borttagningav lignin inte alltid är nödvändigt för att erhålla material medegenskaper som är konkurrenskraftiga till deras fossilbaseradeanaloger.

Place, publisher, year, edition, pages
KTH Royal Institute of Technology, 2023. p. 69
Series
TRITA-CBH-FOU ; 2023:23
Keywords
lignin, biomass valorization, extraction, tissue culture, lignin nanoparticles, wood aerogels, lignin, biomassavalorisering, extraktion, vävnadsodling, lignin nanopartiklar, trä aerogeler
National Category
Wood Science
Research subject
Fibre and Polymer Science
Identifiers
urn:nbn:se:kth:diva-326773 (URN)978-91-8040-589-8 (ISBN)
Public defence
2023-06-14, D3, Lindstedtsvägen 9, Stockholm, 10:00 (English)
Opponent
Supervisors
Note

QC 2023-05-15

Embargo godkänt av tf skolchef Amelie Eriksson Karlström via e-post 2023-05-13

Available from: 2023-05-15 Created: 2023-05-11 Last updated: 2023-06-13Bibliographically approved

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Sapouna, IoannaLawoko, Martin

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