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      Identification of developmental stage and anatomical fraction contributions to cell wall recalcitrance in switchgrass

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          Abstract

          Background

          Heterogeneity within herbaceous biomass can present important challenges for processing feedstocks to cellulosic biofuels. Alterations to cell wall composition and organization during plant growth represent major contributions to heterogeneity within a single species or cultivar. To address this challenge, the focus of this study was to characterize the relationship between composition and properties of the plant cell wall and cell wall response to deconstruction by NaOH pretreatment and enzymatic hydrolysis for anatomical fractions (stem internodes, leaf sheaths, and leaf blades) within switchgrass at various tissue maturities as assessed by differing internode.

          Results

          Substantial differences in both cell wall composition and response to deconstruction were observed as a function of anatomical fraction and tissue maturity. Notably, lignin content increased with tissue maturity concurrently with decreasing ferulate content across all three anatomical fractions. Stem internodes exhibited the highest lignin content as well as the lowest hydrolysis yields, which were inversely correlated to lignin content. Confocal microscopy was used to demonstrate that removal of cell wall aromatics (i.e., lignins and hydroxycinnamates) by NaOH pretreatment was non-uniform across diverse cell types. Non-cellulosic polysaccharides were linked to differences in cell wall response to deconstruction in lower lignin fractions. Specifically, leaf sheath and leaf blade were found to have higher contents of substituted glucuronoarabinoxylans and pectic polysaccharides. Glycome profiling demonstrated that xylan and pectic polysaccharide extractability varied with stem internode maturity, with more mature internodes requiring harsher chemical extractions to remove comparable glycan abundances relative to less mature internodes. While enzymatic hydrolysis was performed on extractives-free biomass, extractible sugars (i.e., starch and sucrose) comprised a significant portion of total dry weight particularly in stem internodes, and may provide an opportunity for recovery during processing.

          Conclusions

          Cell wall structural differences within a single plant can play a significant role in feedstock properties and have the potential to be exploited for improving biomass processability during a biorefining process. The results from this work demonstrate that cell wall lignin content, while generally exhibiting a negative correlation with enzymatic hydrolysis yields, is not the sole contributor to cell wall recalcitrance across diverse anatomical fractions within switchgrass.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13068-017-0870-5) contains supplementary material, which is available to authorized users.

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          Most cited references57

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          Features of promising technologies for pretreatment of lignocellulosic biomass.

          N. Mosier (2005)
          Cellulosic plant material represents an as-of-yet untapped source of fermentable sugars for significant industrial use. Many physio-chemical structural and compositional factors hinder the enzymatic digestibility of cellulose present in lignocellulosic biomass. The goal of any pretreatment technology is to alter or remove structural and compositional impediments to hydrolysis in order to improve the rate of enzyme hydrolysis and increase yields of fermentable sugars from cellulose or hemicellulose. These methods cause physical and/or chemical changes in the plant biomass in order to achieve this result. Experimental investigation of physical changes and chemical reactions that occur during pretreatment is required for the development of effective and mechanistic models that can be used for the rational design of pretreatment processes. Furthermore, pretreatment processing conditions must be tailored to the specific chemical and structural composition of the various, and variable, sources of lignocellulosic biomass. This paper reviews process parameters and their fundamental modes of action for promising pretreatment methods.
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            Hemicelluloses.

            Hemicelluloses are polysaccharides in plant cell walls that have beta-(1-->4)-linked backbones with an equatorial configuration. Hemicelluloses include xyloglucans, xylans, mannans and glucomannans, and beta-(1-->3,1-->4)-glucans. These types of hemicelluloses are present in the cell walls of all terrestrial plants, except for beta-(1-->3,1-->4)-glucans, which are restricted to Poales and a few other groups. The detailed structure of the hemicelluloses and their abundance vary widely between different species and cell types. The most important biological role of hemicelluloses is their contribution to strengthening the cell wall by interaction with cellulose and, in some walls, with lignin. These features are discussed in relation to widely accepted models of the primary wall. Hemicelluloses are synthesized by glycosyltransferases located in the Golgi membranes. Many glycosyltransferases needed for biosynthesis of xyloglucans and mannans are known. In contrast, the biosynthesis of xylans and beta-(1-->3,1-->4)-glucans remains very elusive, and recent studies have led to more questions than answers.
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              Feedstocks for lignocellulosic biofuels.

              In 2008, the world produced approximately 87 gigaliters of liquid biofuels, which is roughly equal to the volume of liquid fuel consumed by Germany that year. Essentially, all of this biofuel was produced from crops developed for food production, raising concerns about the net energy and greenhouse gas effects and potential competition between use of land for production of fuels, food, animal feed, fiber, and ecosystem services. The pending implementation of improved technologies to more effectively convert the nonedible parts of plants (lignocellulose) to liquid fuels opens diverse options to use biofuel feedstocks that reach beyond current crops and the land currently used for food and feed. However, there has been relatively little discussion of what types of plants may be useful as bioenergy crops.
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                Author and article information

                Contributors
                crowejac@msu.edu
                feringan@msu.edu
                spattathil@lallemand.com
                bmerritt1762@gmail.com
                foster54@msu.edu
                dbd@msu.edu
                rgong1@mtu.edu
                hodgeda@msu.edu
                Journal
                Biotechnol Biofuels
                Biotechnol Biofuels
                Biotechnology for Biofuels
                BioMed Central (London )
                1754-6834
                15 July 2017
                15 July 2017
                2017
                : 10
                : 184
                Affiliations
                [1 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Department of Chemical Engineering and Materials Science, , Michigan State University, ; East Lansing, MI USA
                [2 ]ISNI 0000 0004 1936 738X, GRID grid.213876.9, Complex Carbohydrate Research Center, , University of Georgia, ; Athens, GA USA
                [3 ]ISNI 0000 0004 0446 2659, GRID grid.135519.a, Bioenergy Science Center, , Oak Ridge National Laboratory, ; Oak Ridge, TN USA
                [4 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, , DOE-Great Lakes Bioenergy Research Center, Michigan State University, ; East Lansing, MI USA
                [5 ]ISNI 0000 0001 0663 5937, GRID grid.259979.9, Department of Chemical Engineering, , Michigan Technological University, ; Houghton, MI USA
                [6 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Department of Biosystems & Agricultural Engineering, , Michigan State University, ; East Lansing, MI USA
                [7 ]ISNI 0000 0001 1014 8699, GRID grid.6926.b, Department of Civil, Environmental and Natural Resources Engineering, , Luleå University of Technology, ; Luleå, Sweden
                Article
                870
                10.1186/s13068-017-0870-5
                5512841
                28053662
                4eb7d14d-d242-4919-ada3-b13f3fd930df
                © The Author(s) 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 26 May 2017
                : 6 July 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100006206, Biological and Environmental Research;
                Award ID: DE-FC02-07ER64494
                Funded by: FundRef http://dx.doi.org/10.13039/100000146, Division of Chemical, Bioengineering, Environmental, and Transport Systems;
                Award ID: 1336622
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100006132, Office of Science;
                Award ID: DE-AC05-00OR22725
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Biotechnology
                switchgrass,recalcitrance,cell wall glycans,alkaline pretreatment
                Biotechnology
                switchgrass, recalcitrance, cell wall glycans, alkaline pretreatment

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