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      A finalized determinant for complete lignocellulose enzymatic saccharification potential to maximize bioethanol production in bioenergy Miscanthus

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          Abstract

          Background

          Miscanthus is a leading bioenergy crop with enormous lignocellulose production potential for biofuels and chemicals. However, lignocellulose recalcitrance leads to biomass process difficulty for an efficient bioethanol production. Hence, it becomes essential to identify the integrative impact of lignocellulose recalcitrant factors on cellulose accessibility for biomass enzymatic hydrolysis. In this study, we analyzed four typical pairs of Miscanthus accessions that showed distinct cell wall compositions and sorted out three major factors that affected biomass saccharification for maximum bioethanol production.

          Results

          Among the three optimal (i.e., liquid hot water, H 2SO 4 and NaOH) pretreatments performed, mild alkali pretreatment (4% NaOH at 50 °C) led to almost complete biomass saccharification when 1% Tween-80 was co-supplied into enzymatic hydrolysis in the desirable Miscanthus accessions. Consequently, the highest bioethanol yields were obtained at 19% (% dry matter) from yeast fermentation, with much higher sugar–ethanol conversion rates by 94–98%, compared to the other Miscanthus species subjected to stronger pretreatments as reported in previous studies. By comparison, three optimized pretreatments distinctively extracted wall polymers and specifically altered polymer features and inter-linkage styles, but the alkali pretreatment caused much increased biomass porosity than that of the other pretreatments. Based on integrative analyses, excellent equations were generated to precisely estimate hexoses and ethanol yields under various pretreatments and a hypothetical model was proposed to outline an integrative impact on biomass saccharification and bioethanol production subjective to a predominate factor (CR stain) of biomass porosity and four additional minor factors (DY stain, cellulose DP, hemicellulose X/A, lignin G-monomer).

          Conclusion

          Using four pairs of Miscanthus samples with distinct cell wall composition and varied biomass saccharification, this study has determined three main factors of lignocellulose recalcitrance that could be significantly reduced for much-increased biomass porosity upon optimal pretreatments. It has also established a novel standard that should be applicable to judge any types of biomass process technology for high biofuel production in distinct lignocellulose substrates. Hence, this study provides a potential strategy for precise genetic modification of lignocellulose in all bioenergy crops.

          Electronic supplementary material

          The online version of this article (10.1186/s13068-019-1437-4) contains supplementary material, which is available to authorized users.

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          Most cited references42

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          Characterization and genomic analysis of kraft lignin biodegradation by the beta-proteobacterium Cupriavidus basilensis B-8

          Background Lignin materials are abundant and among the most important potential sources for biofuel production. Development of an efficient lignin degradation process has considerable potential for the production of a variety of chemicals, including bioethanol. However, lignin degradation using current methods is inefficient. Given their immense environmental adaptability and biochemical versatility, bacterial could be used as a valuable tool for the rapid degradation of lignin. Kraft lignin (KL) is a polymer by-product of the pulp and paper industry resulting from alkaline sulfide treatment of lignocellulose, and it has been widely used for lignin-related studies. Results Beta-proteobacterium Cupriavidus basilensis B-8 isolated from erosive bamboo slips displayed substantial KL degradation capability. With initial concentrations of 0.5–6 g L-1, at least 31.3% KL could be degraded in 7 days. The maximum degradation rate was 44.4% at the initial concentration of 2 g L-1. The optimum pH and temperature for KL degradation were 7.0 and 30°C, respectively. Manganese peroxidase (MnP) and laccase (Lac) demonstrated their greatest level of activity, 1685.3 U L-1 and 815.6 U L-1, at the third and fourth days, respectively. Many small molecule intermediates were formed during the process of KL degradation, as determined using GC-MS analysis. In order to perform metabolic reconstruction of lignin degradation in this bacterium, a draft genome sequence for C. basilensis B-8 was generated. Genomic analysis focused on the catabolic potential of this bacterium against several lignin-derived compounds. These analyses together with sequence comparisons predicted the existence of three major metabolic pathways: β-ketoadipate, phenol degradation, and gentisate pathways. Conclusion These results confirmed the capability of C. basilensis B-8 to promote KL degradation. Whole genomic sequencing and systematic analysis of the C. basilensis B-8 genome identified degradation steps and intermediates from this bacterial-mediated KL degradation method. Our findings provide a theoretical basis for research into the mechanisms of lignin degradation as well as a practical basis for biofuel production using lignin materials.
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            Hemicelluloses negatively affect lignocellulose crystallinity for high biomass digestibility under NaOH and H2SO4 pretreatments in Miscanthus

            Background Lignocellulose is the most abundant biomass on earth. However, biomass recalcitrance has become a major factor affecting biofuel production. Although cellulose crystallinity significantly influences biomass saccharification, little is known about the impact of three major wall polymers on cellulose crystallization. In this study, we selected six typical pairs of Miscanthus samples that presented different cell wall compositions, and then compared their cellulose crystallinity and biomass digestibility after various chemical pretreatments. Results A Miscanthus sample with a high hemicelluloses level was determined to have a relatively low cellulose crystallinity index (CrI) and enhanced biomass digestibility at similar rates after pretreatments of NaOH and H2SO4 with three concentrations. By contrast, a Miscanthus sample with a high cellulose or lignin level showed increased CrI and low biomass saccharification, particularly after H2SO4 pretreatment. Correlation analysis revealed that the cellulose CrI negatively affected biomass digestion. Increased hemicelluloses level by 25% or decreased cellulose and lignin contents by 31% and 37% were also found to result in increased hexose yields by 1.3-times to 2.2-times released from enzymatic hydrolysis after NaOH or H2SO4 pretreatments. The findings indicated that hemicelluloses were the dominant and positive factor, whereas cellulose and lignin had synergistic and negative effects on biomass digestibility. Conclusions Using six pairs of Miscanthus samples with different cell wall compositions, hemicelluloses were revealed to be the dominant factor that positively determined biomass digestibility after pretreatments with NaOH or H2SO4 by negatively affecting cellulose crystallinity. The results suggested potential approaches to the genetic modifications of bioenergy crops.
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              Investigating plant cell wall components that affect biomass recalcitrance in poplar and switchgrass

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                Author and article information

                Contributors
                alamagrichemist@yahoo.com
                121412336@qq.com
                752624417@qq.com
                hjfjiayoua@yeah.net
                wyt@mail.hzau.edu.cn
                huzhen19900817@163.com
                madadim2002@gmail.com
                sundan510@163.com
                rhuvip@163.com
                aragausk@utk.edu
                yuant@mail.hzau.edu.cn
                +86-27-87281765 , pengliangcai2007@sina.com , lpeng@mail.hzau.edu.cn , http://bbrc.hzau.edu.cn
                Journal
                Biotechnol Biofuels
                Biotechnol Biofuels
                Biotechnology for Biofuels
                BioMed Central (London )
                1754-6834
                27 April 2019
                27 April 2019
                2019
                : 12
                : 99
                Affiliations
                [1 ]ISNI 0000 0004 1790 4137, GRID grid.35155.37, Biomass and Bioenergy Research Centre, , Huazhong Agricultural University, ; Wuhan, 430070 China
                [2 ]ISNI 0000 0004 1790 4137, GRID grid.35155.37, College of Plant Science and Technology, , Huazhong Agricultural University, ; Wuhan, 430070 China
                [3 ]ISNI 0000 0000 8822 034X, GRID grid.411410.1, School of Materials and Chemical Engineering, , Hubei University of Technology, ; Wuhan, 430068 China
                [4 ]ISNI 0000 0004 1759 225X, GRID grid.412979.0, College of Food Science and Technology, , Hubei University of Arts and Science, ; Xiangyang, 441053 China
                [5 ]ISNI 0000 0001 2315 1184, GRID grid.411461.7, Department of Chemical and Biomolecular Engineering, , University of Tennessee-Knoxville, ; Knoxville, TN 37996-2200 USA
                Author information
                http://orcid.org/0000-0002-8739-3470
                Article
                1437
                10.1186/s13068-019-1437-4
                6486690
                f017623e-50fe-4e70-9ecc-a7b0fdc6e416
                © The Author(s) 2019

                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 February 2019
                : 13 April 2019
                Funding
                Funded by: National Key Research and Development Program
                Award ID: 2016YFD0800804
                Award Recipient :
                Funded by: National Science Foundation of China
                Award ID: 31670296
                Award ID: 31571721
                Award Recipient :
                Funded by: National 111 Project of Ministry of Education of China
                Award ID: B08032
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2019

                Biotechnology
                miscanthus,biomass saccharification,bioethanol yield,polymer features,polymer linkages,biomass porosity

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