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      Model-based optimization and scale-up of multi-feed simultaneous saccharification and co-fermentation of steam pre-treated lignocellulose enables high gravity ethanol production

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          Abstract

          Background

          High content of water-insoluble solids (WIS) is required for simultaneous saccharification and co-fermentation (SSCF) operations to reach the high ethanol concentrations that meet the techno-economic requirements of industrial-scale production. The fundamental challenges of such processes are related to the high viscosity and inhibitor contents of the medium. Poor mass transfer and inhibition of the yeast lead to decreased ethanol yield, titre and productivity. In the present work, high-solid SSCF of pre-treated wheat straw was carried out by multi-feed SSCF which is a fed-batch process with additions of substrate, enzymes and cells, integrated with yeast propagation and adaptation on the pre-treatment liquor. The combined feeding strategies were systematically compared and optimized using experiments and simulations.

          Results

          For high-solid SSCF process of SO 2-catalyzed steam pre-treated wheat straw, the boosted solubilisation of WIS achieved by having all enzyme loaded at the beginning of the process is crucial for increased rates of both enzymatic hydrolysis and SSCF. A kinetic model was adapted to simulate the release of sugars during separate hydrolysis as well as during SSCF. Feeding of solid substrate to reach the instantaneous WIS content of 13 % (w/w) was carried out when 60 % of the cellulose was hydrolysed, according to simulation results. With this approach, accumulated WIS additions reached more than 20 % (w/w) without encountering mixing problems in a standard bioreactor. Feeding fresh cells to the SSCF reactor maintained the fermentation activity, which otherwise ceased when the ethanol concentration reached 40–45 g L −1. In lab scale, the optimized multi-feed SSCF produced 57 g L −1 ethanol in 72 h. The process was reproducible and resulted in 52 g L −1 ethanol in 10 m 3 scale at the SP Biorefinery Demo Plant.

          Conclusions

          SSCF of WIS content up to 22 % (w/w) is reproducible and scalable with the multi-feed SSCF configuration and model-aided process design. For simultaneous saccharification and fermentation, the overall efficiency relies on balanced rates of substrate feeding and conversion. Multi-feed SSCF provides the possibilities to balance interdependent rates by systematic optimization of the feeding strategies. The optimization routine presented in this work can easily be adapted for optimization of other lignocellulose-based fermentation systems.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13068-016-0500-7) contains supplementary material, which is available to authorized users.

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          Most cited references23

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          Techno-economic evaluation of producing ethanol from softwood: comparison of SSF and SHF and identification of bottlenecks.

          The aim of the study was to evaluate, from a technical and economic standpoint, the enzymatic processes involved in the production of fuel ethanol from softwood. Two base case configurations, one based on simultaneous saccharification and fermentation (SSF) and one based on separate hydrolysis and fermentation (SHF), were evaluated and compared. The process conditions selected were based mainly on laboratory data, and the processes were simulated by use of Aspen plus. The capital costs were estimated using the Icarus Process Evaluator. The ethanol production costs for the SSF and SHF base cases were 4.81 and 5.32 SEK/L or 0.57 and 0.63 USD/L (1 USD = 8.5SEK), respectively. The main reason for SSF being lower was that the capital cost was lower and the overall ethanol yield was higher. A major drawback of the SSF process is the problem with recirculation of yeast following the SSF step. Major economic improvements in both SSF and SHF could be achieved by increasing the income from the solid fuel coproduct. This is done by lowering the energy consumption in the process through running the enzymatic hydrolysis or the SSF step at a higher substrate concentration and by recycling the process streams. Running SSF with use of 8% rather than 5% nonsoluble solid material would result in a 19% decrease in production cost. If after distillation 60% of the stillage stream was recycled back to the SSF step, the production cost would be reduced by 14%. The cumulative effect of these various improvements was found to result in a production cost of 3.58 SEK/L (0.42 USD/L) for the SSF process.
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            Techno-economic evaluation of bioethanol production from three different lignocellulosic materials

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              Microplate-based filter paper assay to measure total cellulase activity.

              The standard filter paper assay (FPA) published by the International Union of Pure and Applied Chemistry (IUPAC) is widely used to determine total cellulase activity. However, the IUPAC method is not suitable for the parallel analyses of large sample numbers. We describe here a microplate-based method for assaying large sample numbers. To achieve this, we reduced the enzymatic reaction volume to 60 microl from the 1.5 ml used in the IUPAC method. The modified 60-microl format FPA can be carried out in 96-well assay plates. Statistical analyses showed that the cellulase activities of commercial cellulases from Trichoderma reesei and Aspergillus species determined with our 60-microl format FPA were not significantly different from the activities measured with the standard FPA. Our results also indicate that the 60-microl format FPA is quantitative and highly reproducible. Moreover, the addition of excess beta-glucosidase increased the sensitivity of the assay by up to 60%. 2004 Wiley Periodicals, Inc.
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                Author and article information

                Contributors
                ruifei@chalmers.se
                pornkamol.unr@biotec.or.th
                franzen@chalmers.se
                Journal
                Biotechnol Biofuels
                Biotechnol Biofuels
                Biotechnology for Biofuels
                BioMed Central (London )
                1754-6834
                18 April 2016
                18 April 2016
                2016
                : 9
                : 88
                Affiliations
                [ ]Division of Industrial Biotechnology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden
                [ ]National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani, Thailand
                Article
                500
                10.1186/s13068-016-0500-7
                4835939
                27096006
                2db3fd64-dd17-439a-b5cb-5cfc91f77497
                © Wang et al. 2016

                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
                : 3 February 2016
                : 1 April 2016
                Funding
                Funded by: Energimyndigheten (SE)
                Award ID: P37353-1
                Award Recipient :
                Funded by: Chalmers Energy Inititative
                Categories
                Research
                Custom metadata
                © The Author(s) 2016

                Biotechnology
                biofuels,fermentation technology,agricultural residues,bioprocessing,enzymatic hydrolysis,high gravity,demo-scale simultaneous saccharification and fermentation,mathematical modelling,fed-batch ssf

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