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  • Periodic Reporting for period 1 - US4GREENCHEM (Combined Ultrasonic and Enzyme treatment of Lignocellulosic Feedstock as Substrate for Sugar Based Biotechnological Applications)

US4GREENCHEM Report Summary

Project ID: 669055
Funded under: H2020-EU.3.2.6.

Periodic Reporting for period 1 - US4GREENCHEM (Combined Ultrasonic and Enzyme treatment of Lignocellulosic Feedstock as Substrate for Sugar Based Biotechnological Applications)

Reporting period: 2015-07-01 to 2016-12-31

Summary of the context and overall objectives of the project

Rising concern over fossil fuels supply and the impacts of GHG have led to high interest in new feedstocks for manufacturing industries. Using biomass as sustainable renewable resource is a way to replace carbon from fossil sources in the manufacture of chemicals, materials and fuels.
Due to the limited arable surface of Europe, the major potential for European biorefineries lies with lignocellulosic substrates derived from agriculture and forestry residues and from energy crops (SRC).
Objectives: US4GREENCHEM aims to design a biorefinery concept for the complete valorization of lignocellulosic biomass that is energy- and cost- efficient and based on green technologies.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

WP1: Mechanical Disruption
Task 1.1: pre-requisites for cavitation
TTZ acquired the biomass (hardwood chips and straw), performed a chemical characterization, and sent samples of the biomass to different milling institutes in Germany to compare the effect of different milling techniques. IWC carried out the surface characterization and showed the difference in the pore structure. Analytical pyrolysis method was applied (D1.1).
Task 1.2: optimization of cavitation parameters
Test tanks were manufactured at WU and distributed to partner TTZ and UNITO for lab scale sonication. WU developed sensors for cavitation activity and for the scaled-up prototype in WP 6. Requirements of the sensor were determined, initial measurements were performed, and a tentative flexible system design was implemented. UNITO sieved milled biomass. The influence of particle size on the sonication pre-treatment was evaluated and compared to conventional high speed stirring.
Task 1.3: impact of cavitation on biomass structure
Ultrasound treated samples (UNITO) analyzed at IWC for surface characterization using nitrogen gas adsorption-desorption isotherms, obtaining results on total pore/micropore volume/width, specific surface area, and size distribution. Samples before and after US treatment analyzed using FTIR and EPR spectroscopy. Formation of free radical resulting from US pre-treatment detected (D1.5). Further pretreatment with ultrasonication and hydrodynamic cavitation in 10 % sodium hydroxide(UNITO).
WP2: Matrix Disintegration
Task 2.1: CO2 Hemicellulose Hydrolysis
FeyeCon performed subcritical and supercritical carbon dioxide hydrolysis at a high pressure, temperature, and long pre-treatment time. 75 prepared sample were sent to VTT, BC, and IWC for the analysis. Experimental work was proceeded according the analysis results.
Task 2.2: High frequency sonochemical disruption
Based on the existing lab-scale tank, a high-frequency transducer was developed for biomass treatment at UNITO with WU contribution. Wheat straw samples were sonicated at 500 kHz. The results show a significantly lower mechanical effect on the samples caused by cavitation.
Task 2.3: Integration of the proposed technologies
WU designed an ultrasound probe (40 kHz) and installed into existing Sc-CO2 extraction unit. integrated US- supercritical CO2 (ScCO2) tests were carried out. A specialized transducer design was needed to integrate ultrasonic treatment into an existing high-pressure vessel at FEYECON.
Task 2.4: Characterization of the lignin-derived fractions
IWC has analyzed 72 solid and 20 liquid samples and FeyeCon characterized lignin-derived fractions of pre-treated biomass. The modification of chemical structure of lignin and cellulose biomass constituents under influence of combined US and Sc-CO2 pre-treatment was revealed. (D2.4 and D2.5)
WP3: Biocatalysis Design
Task 3.1: Enzymatic Digestibility studies
Samples tested with enzymatic hydrolysis at VTT using commercial FlashZymes and at Biocentras using enzyme biosynthesized by Trichoderma reesei strain 101, as reported in D3.1. The best soluble sugar yields reached, both at VTT and at BC, in enzymatic hydrolysis of the wheat straw sample pre-treated with ultrasonication in sodium hydroxide.
Task 3.2: New enzymes for improved hydrolysis
Three novel fungal beta-glucosidases cloned at VTT are being examined as candidates for completing the VTT enzyme mixture. 4 novel genes coding for beta-glucosidases have been identified from VTT’s in-house RNA sequencing database. Experiments for characterization of lignocellulolytic enzymes from Trichoderma reesei 101 strain initialized (BC)
Task 3.3: Increase of enzyme yields
Acreening of fungal strains from Biocentras culture collection has been started to increase yields of enzymes.
WP4: Sugar Recovery
Task4.1: Optimized Hydrolysis
The hydrolysis optimization is in progress on sample 21 at VTT. Efforts focused on search for greener pre-treatment conditions.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

US4GREENCHEM is researching on optimizing 3 major steps of a state-of-the-art biorefinery for conversion pathway of lignocellulosic substrates into biofuel and biomaterials, reducing processing costs: physical pre-treatment with ultrasound and supercritical CO2 technologies, a multi-target hydrolysis and valorization of process effluents.
Socio-economic impacts include knowledge creation, increased academia-industry cooperation, creation of new markets, reutilization of local residues and support the regional development, valorization of agricultural residues, reducing dependence on imports of fossil oil, new skilled jobs in rural regions and in product development and engineering.

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