Phragmites australis (common reed) is a perennial grass that grows in wetlands or near inland waterways. Due to its fast-growing properties and low requirement in nutrients and water, this arboreal variety is recognized as a promising source of renewable energy although it is one of the least characterized energy crops. In this experiment, the optimization of the bioethanol production process from Phragmites australis was carried out. Raw material was first characterized according to the standard procedure (NREL) to evaluate its composition in terms of cellulose, hemicellulose, and lignin content. Common reed was pretreated by steam explosion process at three different severity factor (R₀) values. The pretreatment was performed in order to reduce biomass recalcitrance and to make cellulose more accessible to enzymatic attack. After the pretreatment, a water insoluble substrate (WIS) rich in cellulose and lignin and a liquid fraction rich in pentose sugars (xylose and arabinose) and inhibitors were collected and analyzed. The simultaneous saccharification and fermentation (SSF) of the WIS was performed at three different solid loadings (SL) 10%, 15%, 20% (w/w). The same enzyme dosage, equal to 20% (g enzyme/g cellulose), was used for all the WIS loadings. The efficiency of the whole process was evaluated in terms of ethanol overall yield (g ethanol/100 g raw material). The maximum ethanol overall yields achieved were 16.56 and 15.80 g ethanol/100 g RM dry basis for sample AP10 and sample AP4.4, respectively. The yields were reached working at lower solid loading (10%) and at the intermediate LogR₀ value for the former and at intermediate solid loading (15%) and high LogR₀ value for the latter, respectively.

Some lignocellulosic biomass feedstocks occur in Mediterranean Countries. They are still largely unexploited and cause considerable problems due to the lack of cost-effective harvesting, storage and disposal technologies. Recent studies found that some basidiomycetous yeasts are able to accumulate high amount of intracellular lipids for biorefinery processes (i.e., biofuels and biochemicals). Accordingly, the above biomass feedstocks could be used as carbon sources (after their pre-treatment and hydrolysis) for lipid accumulation by oleaginous yeasts. Cardoon stalks, stranded driftwood and olive tree pruning residues were pre-treated with steam-explosion and enzymatic hydrolysis for releasing free mono- and oligosaccharides. Lipid accumulation tests were performed at two temperatures (20 and 25 °C) using Leucosporidium creatinivorum DBVPG 4794, Naganishia adeliensis DBVPG 5195 and Solicoccozyma terricola DBVPG 5870. S. terricola grown on cardoon stalks at 20 °C exhibited the highest lipid production (13.20 g/l), a lipid yield (28.95%) close to the maximum theoretical value and a lipid composition similar to that found in palm oil. On the contrary, N. adeliensis grown on stranded driftwood and olive tree pruning residues exhibited a lipid composition similar to those of olive and almonds oils. A predictive evaluation of the physical properties of the potential biodiesel obtainable by lipids produced by tested yeast strains has been reported and discussed. Lipids produced by some basidiomycetous yeasts grown on Mediterranean lignocellulosic biomass feedstocks could be used as supplementary sources of oils for producing biofuels and biochemicals.

One of the primary targets for the new lignocellulosic feedstock-based biorefinery is the simultaneous valorization of holocellulose and lignin. Acidified organosolv treatment is among the most promising strategy for recovering technical lignin, water-soluble hemicellulose, and cellulose pulp with increased accessibility to hydrolytic enzymes. In this work, a design-of-experiment (DoE) approach was used to increase the cellulose recovery, digestibility, and the delignification of Cynara cardunculus L. feedstock. In the first treatment, the milled biomass was subjected to microwave-assisted extraction using an acidified GVL/water mixture to separate lignin and hemicellulose from cellulose. In the second treatment, the cellulose pulp was hydrolyzed by cellulolytic enzymes to demonstrate the enhanced digestibility. At the optimal condition (154 °C, 2.24% H₂SO₄, and 0.62 GVL/water ratio), the cellulose pulp showed a cellulose content of 87.59%, while the lignin content was lower than 8%. The cellulose recovery and digestibility were equal to 79.46% and 86.94%, respectively. About 40% of the initial hemicellulose was recovered as monosaccharides. This study demonstrated the effectiveness of the two-step organosolv treatment for biomass fractionation; however, as suggested by DoE analysis, a confirmative study at a low temperature (<154 °C) should be performed to further increase the cellulose recovery.

Biorefinery aims at designing new virtuous and high-efficiency energy chains, achieving the combined production of biofuels (e.g., bioethanol) and biobased products. This emerging philosophy can represent an important opportunity for the industrial world, exploiting a new kind of nano-smart biomaterials in their production chains. This paper will present the lab experience carried out by the Biomass Research Centre (CRB) in extracting cellulose nanocrystals (NCC) from a pretreated (via Steam Explosion) fraction of Cynara cardunculus. This is a very common and invasive arboreal variety in central Italy. The NCC extraction methodology allows the separation of the crystalline content of cellulose. Such a procedure has been considered in the literature with the exception of one step in which the conditions have been optimized by CRB Lab. This procedure has been applied for the production of NCC from both Cynara cardunculus and microcrystalline cellulose (MCC). The paper will discuss some of the results achieved using the obtained nanocrystals as reinforcing filler in a paper sheet; it was found that the tensile strength increased from 3.69 kg/15 mm to 3.98 kg/15 mm, the durability behavior (measured by bending number) changed from the value 95 to the value 141, and the barrier properties (measured by Gurley porosity) were improved, increasing from 38 s to 45 s.

Cardoon (Cynara cardunculus) is a promising crop from which to obtain oilseeds and lignocellulosic biomass. Acid-catalyzed steam explosion is a thermochemical process that can efficiently pre-treat lignocellulosic biomass. The drawback is the production of a high number of carbohydrate degradation products in the liquid fraction that could inhibit microbial growth. In this work, the lignocellulosic biomass of cardoon, gathered from a dedicated field, were used as the raw material for the production of fermentable monosaccharides by employing acid-catalyzed steam explosion. The raw material was pre-soaked with a dilute 1% (w/w) sulfuric acid solution and then subjected to steam explosion under three different severity conditions. The recovered slurry was separated into solid and liquid fractions, which were individually characterized to determine total carbohydrate and inhibitor concentrations. The slurry and the washed solid fraction underwent enzymatic hydrolysis to release glucose and pentose monosaccharides. By conducting the pre-treatment at 175 °C for 35 min and hydrolyzing the obtained slurry, a yield of 33.17 g of monosaccharides/100 g of cardoon was achieved. At the same conditions, 4.39 g of inhibitors/100 g of cardoon were produced.

Lignocellulosic biomass is a nonedible matrix that can be efficiently exploited as feedstock in an integrated biorefinery after a proper pretreatment. An organosolv pretreatment using an acidified γ-valerolactone (GVL)/water solution was proposed to improve the cellulose enrichment and enzymatic saccharification of cardoon (Cynara cardunculus L.) stems. At the optimal pretreatment condition (140 °C, 0.6 GVL/water, and 2.24% H₂SO₄), xylan was efficiently removed from the cardoon, and up to 50% of its content was recovered in the aqueous fraction, while 86% of the cellulose was retained in the solid fraction. The resulting cardoon pulp showed a cellulose content of 91.5% and an enzymatic digestibility of 100%. An overall glucose production of 37.17 g/100 g raw material (90% theoretical maximum) was obtained using high solid loading (20% w/w) and a high enzyme dosage (60 FPU/g cellulose). At a low enzyme dosage, glucose concentrations of 169 g/L and 210 g/L were achieved using 10 FPU/g cellulose and 20 FPU/g cellulose, respectively. Therefore, an organosolv pretreatment can be an effective process for producing cellulose-enriched pulp with enhanced enzymatic digestibility from cardoon stems, providing a promising option for green lignocellulosic biorefineries that aim to produce high concentrations of glucose with low cellulase addition.