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Enzymatic Degradation of Maize Shoots: Monitoring of Chemical and Physical Changes Reveals Different Saccharification Behaviors
Fabienne Guillon
INRAE: Institut National de Recherche pour l'Agriculture l'Alimentation et l'Environnement
Corresponding Author
ORCiD: https://orcid.org/0000-0001-7925-3104
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This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Biotechnology for Biofuels.
Background
The recalcitrance of lignocellulosics to enzymatic saccharification has been related to many factors, including the tissue and molecular heterogeneity of the plant particles. The role of tissue heterogeneity generally assessed from plant sections is not easy to study on a large scale. In the present work, dry fractionation of ground maize shoot was performed to obtain particle fractions enriched in specific tissue or molecules. The degradation profiles of the fractions were compared considering physical changes in addition to chemical conversion.
Results
Coarse, medium and fine fractions were produced using a dry process followed by an electrostatic separation. The physical and chemical characteristics of the fractions varied, suggesting enrichment in tissue from leaves, pith or rind. The fractions were subjected to seven hours of enzymatic hydrolysis in a torus reactor designed for real-time monitoring of the number and size of the particles. Saccharification efficiency was monitored by analyzing the sugar content of hydrolysate samples collected at different times. The lowest and highest saccharification yields were measured in the coarse and fine fractions, respectively, and these yields paralleled the reduction in the size and number of particles. The behavior of the positively- and negatively-charged particles of medium-size fractions was contrasted. Although the amount of sugar release was similar, the changes in particle size and number differed during enzymatic degradation. The reduction in the number of particles proceeded faster than that of particle size, suggesting that degradable particles were degraded to the point of disappearance with no significant erosion or fragmentation. Considering all fractions, the saccharification yield was positively correlated with the amount of water associated with [5-15 nm] pore size range at 67% moisture content while the reduction in the number of particles was inversely correlated with the amount of lignin. Lignin could act in maintaining the macroscopic structure of particles without necessarily prohibitive effects on the accessibility of enzymes to their substrates.
Conclusion
Real-time monitoring of sugar release and changes in the number and size of the particles clearly evidenced different degradation patterns for fractions of maize shoot that could be related to tissue heterogeneity in the plant.
Keywords
lignocellulosic, plant dry fractionation, recalcitrance, particles size, image analysis, time-lapse study, porosity
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CURRENT STATUS: Published
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Posted 18 Sep, 2020
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On 05 Jan, 2021
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Editorial decision: Major revision
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Received 30 Oct, 2020
Reviewer #2 agreed
On 13 Oct, 2020
Review #1 received
Received 08 Oct, 2020
Reviewer #1 agreed
On 24 Sep, 2020
Reviewers invited
Invitations sent on 23 Sep, 2020
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On 16 Sep, 2020
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On 15 Sep, 2020
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This preprint is under consideration at Biotechnology for Biofuels. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Enzymatic Degradation of Maize Shoots: Monitoring of Chemical and Physical Changes Reveals Different Saccharification Behaviors
Fabienne Guillon
INRAE: Institut National de Recherche pour l'Agriculture l'Alimentation et l'Environnement
Corresponding Author
ORCiD: https://orcid.org/0000-0001-7925-3104
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 1
Posted 18 Sep, 2020
Published
On 05 Jan, 2021
No community comments so far
Editorial decision: Major revision
On 04 Nov, 2020
Review #2 received
Received 30 Oct, 2020
Reviewer #2 agreed
On 13 Oct, 2020
Review #1 received
Received 08 Oct, 2020
Reviewer #1 agreed
On 24 Sep, 2020
Reviewers invited
Invitations sent on 23 Sep, 2020
Editor assigned
On 16 Sep, 2020
First submitted
On 15 Sep, 2020
Submission checks complete
On 15 Sep, 2020
Editor invited
On 15 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Biotechnology and Bioengineering
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Biotechnology for Biofuels.
Background
The recalcitrance of lignocellulosics to enzymatic saccharification has been related to many factors, including the tissue and molecular heterogeneity of the plant particles. The role of tissue heterogeneity generally assessed from plant sections is not easy to study on a large scale. In the present work, dry fractionation of ground maize shoot was performed to obtain particle fractions enriched in specific tissue or molecules. The degradation profiles of the fractions were compared considering physical changes in addition to chemical conversion.
Results
Coarse, medium and fine fractions were produced using a dry process followed by an electrostatic separation. The physical and chemical characteristics of the fractions varied, suggesting enrichment in tissue from leaves, pith or rind. The fractions were subjected to seven hours of enzymatic hydrolysis in a torus reactor designed for real-time monitoring of the number and size of the particles. Saccharification efficiency was monitored by analyzing the sugar content of hydrolysate samples collected at different times. The lowest and highest saccharification yields were measured in the coarse and fine fractions, respectively, and these yields paralleled the reduction in the size and number of particles. The behavior of the positively- and negatively-charged particles of medium-size fractions was contrasted. Although the amount of sugar release was similar, the changes in particle size and number differed during enzymatic degradation. The reduction in the number of particles proceeded faster than that of particle size, suggesting that degradable particles were degraded to the point of disappearance with no significant erosion or fragmentation. Considering all fractions, the saccharification yield was positively correlated with the amount of water associated with [5-15 nm] pore size range at 67% moisture content while the reduction in the number of particles was inversely correlated with the amount of lignin. Lignin could act in maintaining the macroscopic structure of particles without necessarily prohibitive effects on the accessibility of enzymes to their substrates.
Conclusion
Real-time monitoring of sugar release and changes in the number and size of the particles clearly evidenced different degradation patterns for fractions of maize shoot that could be related to tissue heterogeneity in the plant.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10