Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes.
Identifieur interne : 001397 ( Main/Exploration ); précédent : 001396; suivant : 001398Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes.
Auteurs : Gabriel Paës [France] ; Anouck Habrant [France] ; Jordane Ossemond [France] ; Brigitte Chabbert [France]Source :
- Biotechnology for biofuels [ 1754-6834 ] ; 2017.
Abstract
BACKGROUND
The lignocellulosic cell wall network is resistant to enzymatic degradation due to the complex chemical and structural features. Pretreatments are thus commonly used to overcome natural recalcitrance of lignocellulose. Characterization of their impact on architecture requires combinatory approaches. However, the accessibility of the lignocellulosic cell walls still needs further insights to provide relevant information.
RESULTS
Poplar specimens were pretreated using different conditions. Chemical, spectral, microscopic and immunolabeling analysis revealed that poplar cell walls were more altered by sodium chlorite-acetic acid and hydrothermal pretreatments but weakly modified by soaking in aqueous ammonium. In order to evaluate the accessibility of the pretreated poplar samples, two fluorescent probes (rhodamine B-isothiocyanate-dextrans of 20 and 70 kDa) were selected, and their mobility was measured by using the fluorescence recovery after photobleaching (FRAP) technique in a full factorial experiment. The mobility of the probes was dependent on the pretreatment type, the cell wall localization (secondary cell wall and cell corner middle lamella) and the probe size. Overall, combinatory analysis of pretreated poplar samples showed that even the partial removal of hemicellulose contributed to facilitate the accessibility to the fluorescent probes. On the contrary, nearly complete removal of lignin was detrimental to accessibility due to the possible cellulose-hemicellulose collapse.
CONCLUSIONS
Evaluation of plant cell wall accessibility through FRAP measurement brings further insights into the impact of physicochemical pretreatments on lignocellulosic samples in combination with chemical and histochemical analysis. This technique thus represents a relevant approach to better understand the effect of pretreatments on lignocellulose architecture, while considering different limitations as non-specific interactions and enzyme efficiency.
DOI: 10.1186/s13068-017-0704-5
PubMed: 28101142
PubMed Central: PMC5237506
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laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims</wicri:regionArea><placeName><region type="region" nuts="2">Grand Est</region><region type="old region" nuts="2">Champagne-Ardenne</region><settlement type="city">Reims</settlement></placeName><orgName type="university">Université de Reims Champagne-Ardenne</orgName></affiliation></author><author><name sortKey="Habrant, Anouck" sort="Habrant, Anouck" uniqKey="Habrant A" first="Anouck" last="Habrant">Anouck Habrant</name><affiliation wicri:level="4"><nlm:affiliation>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims</wicri:regionArea><placeName><region type="region" nuts="2">Grand 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Brigitte" uniqKey="Chabbert B" first="Brigitte" last="Chabbert">Brigitte Chabbert</name><affiliation wicri:level="4"><nlm:affiliation>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims</wicri:regionArea><placeName><region type="region" nuts="2">Grand Est</region><region type="old region" nuts="2">Champagne-Ardenne</region><settlement type="city">Reims</settlement></placeName><orgName type="university">Université de Reims Champagne-Ardenne</orgName></affiliation></author></analytic><series><title level="j">Biotechnology for biofuels</title><idno type="ISSN">1754-6834</idno><imprint><date when="2017" type="published">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>The lignocellulosic cell wall network is resistant to enzymatic degradation due to the complex chemical and structural features. Pretreatments are thus commonly used to overcome natural recalcitrance of lignocellulose. Characterization of their impact on architecture requires combinatory approaches. However, the accessibility of the lignocellulosic cell walls still needs further insights to provide relevant information.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>Poplar specimens were pretreated using different conditions. Chemical, spectral, microscopic and immunolabeling analysis revealed that poplar cell walls were more altered by sodium chlorite-acetic acid and hydrothermal pretreatments but weakly modified by soaking in aqueous ammonium. In order to evaluate the accessibility of the pretreated poplar samples, two fluorescent probes (rhodamine B-isothiocyanate-dextrans of 20 and 70 kDa) were selected, and their mobility was measured by using the fluorescence recovery after photobleaching (FRAP) technique in a full factorial experiment. The mobility of the probes was dependent on the pretreatment type, the cell wall localization (secondary cell wall and cell corner middle lamella) and the probe size. Overall, combinatory analysis of pretreated poplar samples showed that even the partial removal of hemicellulose contributed to facilitate the accessibility to the fluorescent probes. On the contrary, nearly complete removal of lignin was detrimental to accessibility due to the possible cellulose-hemicellulose collapse.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSIONS</b></p><p>Evaluation of plant cell wall accessibility through FRAP measurement brings further insights into the impact of physicochemical pretreatments on lignocellulosic samples in combination with chemical and histochemical analysis. This technique thus represents a relevant approach to better understand the effect of pretreatments on lignocellulose architecture, while considering different limitations as non-specific interactions and enzyme efficiency.</p></div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">28101142</PMID><DateRevised><Year>2020</Year><Month>10</Month><Day>01</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">1754-6834</ISSN><JournalIssue CitedMedium="Print"><Volume>10</Volume><PubDate><Year>2017</Year></PubDate></JournalIssue><Title>Biotechnology for biofuels</Title><ISOAbbreviation>Biotechnol Biofuels</ISOAbbreviation></Journal><ArticleTitle>Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes.</ArticleTitle><Pagination><MedlinePgn>15</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1186/s13068-017-0704-5</ELocationID><Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">The lignocellulosic cell wall network is resistant to enzymatic degradation due to the complex chemical and structural features. Pretreatments are thus commonly used to overcome natural recalcitrance of lignocellulose. Characterization of their impact on architecture requires combinatory approaches. However, the accessibility of the lignocellulosic cell walls still needs further insights to provide relevant information.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">Poplar specimens were pretreated using different conditions. Chemical, spectral, microscopic and immunolabeling analysis revealed that poplar cell walls were more altered by sodium chlorite-acetic acid and hydrothermal pretreatments but weakly modified by soaking in aqueous ammonium. In order to evaluate the accessibility of the pretreated poplar samples, two fluorescent probes (rhodamine B-isothiocyanate-dextrans of 20 and 70 kDa) were selected, and their mobility was measured by using the fluorescence recovery after photobleaching (FRAP) technique in a full factorial experiment. The mobility of the probes was dependent on the pretreatment type, the cell wall localization (secondary cell wall and cell corner middle lamella) and the probe size. Overall, combinatory analysis of pretreated poplar samples showed that even the partial removal of hemicellulose contributed to facilitate the accessibility to the fluorescent probes. On the contrary, nearly complete removal of lignin was detrimental to accessibility due to the possible cellulose-hemicellulose collapse.</AbstractText><AbstractText Label="CONCLUSIONS" NlmCategory="CONCLUSIONS">Evaluation of plant cell wall accessibility through FRAP measurement brings further insights into the impact of physicochemical pretreatments on lignocellulosic samples in combination with chemical and histochemical analysis. This technique thus represents a relevant approach to better understand the effect of pretreatments on lignocellulose architecture, while considering different limitations as non-specific interactions and enzyme efficiency.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Paës</LastName><ForeName>Gabriel</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Habrant</LastName><ForeName>Anouck</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ossemond</LastName><ForeName>Jordane</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>FARE laboratory, INRA, Université de Reims Champagne-Ardenne, 51100 Reims, 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