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Microplate-Based Evaluation of the Sugar Yield from Giant Reed, Giant Miscanthus and Switchgrass after Mild Chemical Pre-Treatments and Hydrolysis with Tailored Trichoderma Enzymatic Blends

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Abstract

Giant reed, miscanthus, and switchgrass are considered prominent lignocellulosic feedstocks to obtain fermentable sugars for biofuel production. The bioconversion into sugars requires a delignifying pre-treatment step followed by hydrolysis with cellulase and other accessory enzymes like xylanase, especially in the case of alkali pre-treatments, which retain the hemicellulose fraction. Blends richer in accessory enzymes than commercial mix can be obtained growing fungi on feedstock-based substrates, thus ten selected Trichoderma isolates, including the hypercellulolytic strain Trichoderma reesei Rut-C30, were grown on giant reed, miscanthus, or switchgrass-based substrates. The produced enzymes were used to saccharify the corresponding feedstocks, compared to a commercial enzymatic mix (6 FPU/g). Feedstocks were acid (H2SO4 0.2–2%, w/v) or alkali (NaOH 0.02–0.2%, w/v) pre-treated. A microplate-based approach was chosen for most of the experimental steps due to the large number of samples. The highest bioconversion was generally obtained with Trichoderma harzianum Or4/99 enzymes (78, 89, and 94% final sugar yields at 48 h for giant reed, miscanthus, and switchgrass, respectively), with significant increases compared to the commercial mix, especially with alkaline pre-treatments. The differences in bioconversion yields were only partially caused by xylanases (maximum R 2 = 0.5), indicating a role for other accessory enzymes.

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Acknowledgments

This research was carried out partially in the framework of the “BIOSEGEN Project” funded by the Italian Ministry of Agricultural, Food and Forestry Policies, D.M. 17532/7303/10, and partially in the framework of the “AGROENER” project funded by the same Ministry, D.M. 26329/ 2016. The authors gratefully acknowledge Prof. Andrea Monti (University of Bologna), Dr. Enrico Ceotto, and Dr. Mario Di Candilo (CREA) for providing samples of the feedstocks studied, Dr. Ciro Vasmara (CREA) for determining their fiber composition, and Dr. Pier Luigi Burzi (CREA) for the technical assistance in laboratory.

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  1. Council for Agricultural Research and Economics, Research Center for Agriculture and Environment (CREA-AA), Via di Corticella 133, 40128, Bologna, Italy

    Stefano Cianchetta, Luca Bregoli & Stefania Galletti

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  1. Stefano Cianchetta
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Cianchetta, S., Bregoli, L. & Galletti, S. Microplate-Based Evaluation of the Sugar Yield from Giant Reed, Giant Miscanthus and Switchgrass after Mild Chemical Pre-Treatments and Hydrolysis with Tailored Trichoderma Enzymatic Blends. Appl Biochem Biotechnol 183, 876–892 (2017). https://doi.org/10.1007/s12010-017-2470-z

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