• Energy Research

Agricultural by-products such as corn stover are considered strategic raw materials for the production of second-generation bioethanol from renewable and non-food sources. This paper describes the conversion of steam-pretreated corn stover to ethanol utilising a multi-step process including enzymatic hydrolysis, isomerisation, and fermentation of mixed hydrolysates with native Saccharomyces cerevisiae. An immobilised isomerase enzyme was used for the xylose isomerisation along with high concentrations of S. cerevisiae. The objective was to assess the extent of simultaneity of the various conversion steps, through a detailed analysis of process time courses, and to test this process scheme for the conversion of lignocellulosic hydrolysates containing several inhibitors of the isomerase enzyme (e.g. metal ions, xylitol and glycerol). The process was tested on two types of hydrolysate after acid-catalysed steam pretreatment: (a) the water soluble fraction (WSF) in which xylose was the largest carbon source and (b) the entire slurry, containing both cellulose and hemicellulose carbohydrates, in which glucose predominated. The results indicated that the ethanol concentration rose when the inoculum concentration was increased in the range 10-75 g/L. However, when xylose was the largest carbon source, the metabolic yields were higher than 0.51g(ethanol)/g(consumed) sugars probably due to the use of yeast internal cellular resources. This phenomenon was not observed in the fermentation of mixed hydrolysates obtained from the entire pretreated product and in which glucose was the largest carbon source. The ethanol yield from biomass suspensions with dry matter (DM) concentrations of 11-12% (w/v) was 70% based on total sugars (glucose, xylose, galactose). The results suggest that xylulose uptake was more effective in mixed hydrolysates containing glucose levels similar to, or higher than, xylose. Analysis of the factors that limit isomerase activity in lignocellulosic hydrolysates excluded any inhibition due to residual calcium ions after the detoxification of the hemicellulose hydrolysates with Ca(OH)2. By contrast, most of the enzyme activity ceased during the fermentation of the entire slurry after steam explosion, probably due to synergistic inhibition effects of various fermentation co-products.

Abstract Fuel grade bioethanol can be produced from a number of lignocellulosic biomass sources. In the present paper the enzymatic hydrolysis of giant reed ( Arundo donax ) at high biomass loadings up to 20% (w/v) was investigated. Two pretreatment technologies were used: one step steam explosion (SE) with and without acid catalyst and the novel pilot process from Chemtex consisting in a two steps pretreatment. The results of the biomass composition after different pretreatment conditions along with the corresponding enzymatic hydrolizability at low solids loading, indicated that the best pretreatments were the acid catalyzed steam explosion (ACSE) at 200 °C and the Chemtex process in which the SE step was carried out at 206 for 4 min. Various process configurations, including biomass and enzymes fed-batch, were investigated to avoid excessive viscosities and increase the cellulose conversion. Novel enzymatic preparations from Novozymes, Cellic™ Ctec1 and Cellic™ Ctec2, were used for the biomass saccharification. Concerning the analysis of the factors that limited the hydrolysis yields at high DM content, some evidences indicated that product inhibition by xylooligomers could have an important role. The highest solids loading investigated in the present paper was 21%. Under these conditions and by using an enzymes loading of 0.28 g Ctec2/g glucan, an overall sugars concentration of 91 g/L (glucose + xylose) can be obtained after 48 h from ACSE at 200 °C.

Abstract The production of seed oils from Cynara cardunculus generates huge amounts of lignocellulosic residues which can be exploited according to a cascade approach. In this paper, residual cardoon biomass (RCB) was tested as a growth substrate for the solid-state production of cellulolytic cocktails by species known to produce glucose-tolerant β-glucosidase isoenzymes. Best productions were obtained with 10-d-old Aspergillus tubingensis cultures on RCB supplemented with wheat bran (200 g kg−1) yielding β-glucosidase and endo-β-1,4-glucanase activities as high as (25 and 4) IU g−1, respectively, and 4 FPU g−1. The saccharification performance of the obtained cocktail tested on acid-catalysed steam-exploded RCB at low solid loading (25 g dm−3) was around 53% at 20 FPU g−1 cellulose. These performance were significantly enhanced by adding the xylanase-rich NS 22083 commercial formulation, reaching glucose yields higher than 80% after 72 h incubation. The use of the catalytic additive was optimized by a statistical approach, based on factorial analysis. A comparison of the performance of the A. tubingensis reinforced cocktail with the Cellic®CTec2 taken as benchmark formulation was done at the same enzyme load and performed at industrially relevant solid loadings, namely at (100 and 200) g dm−3. This comparison showed that Cellic®CTec2 led to only slightly higher glucose yields while an opposite outcome was observed for xylose yields, irrespective of the solid loading conditions. Thus, this study shows that an in-house enzyme production, based on the solid-state conversion of an industrial byproduct, able of yielding cellulolytic cocktails with substantial saccharification performance is feasible.

The obtainment of sugars from lignocellulosic residues represents a sustainable and versatile platform for the production of a number of bio-based products. Cellulases are a family of enzymes which can effectively hydrolyze the biomass polysaccharides at mild conditions. Cellulolytic fungi belonging to the genera Trichoderma and Aspergillus are the most commonly source of commercial cellulases used so far. More recently, Talaromyces cellulolyticus was also scored as a promising cellulases producer. In comparison to the Trichoderma and Aspergillus systems, Talaromyces enzymes have been less investigated. The present research dealt with the conversion of steam-pretreated corn stover by commercial blend of T. cellulolyticus enzymes with respect to the common blends. The paper also investigated the stability of the enzyme preparation and tested the use of additives (namely Tween 80, Tween 20, and BSA) to improve the enzymes performances and the hydrolysis efficiency. The results indicated that, at the same process conditions, T. cellulolyticus cellulases were more effective and yielded 20% more sugars compared to control blends. Furthermore, the cellulase components displayed a synergistic interaction with hemicellulases. The results indicate that cellulases from T. cellulolyticus are less affected by the high dry matter consistency and the use of additives could increase the total activity by around 50% and β-glucosidase capacity by 10-15%.

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, Arundo donax L. is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast Cutaneotrichosporon curvatus from undetoxified lignocellulosic hydrolysates of steam-pretreated A. donax. The growth of C. curvatus was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of A. donax hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty acid profile of C. curvatus triglycerides contained 27% palmitic acid, 33% oleic acid and 32% linoleic acid. These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Lignocellulosic biomass is currently widely used in many biorefining processes. The full exploitation of biomass from uncultivated or even marginal lands for the production of biobased chemicals has deserved huge attention in the last few years. Among the sustainable biomass-based value chains, cardoon crops could be a feedstock for biorefineries as they can grow on marginal lands and be used as raw material for multipurpose exploitation, including seeds, roots, and epigeous lignocellulosic solid residue. This work focused on the technical analysis of a novel integrated flowsheet for the exploitation of the lignocellulosic fraction through the assessment of thermochemical, biochemical, and extractive technologies and processes. In particular, high-yield thermochemical processes (gasification), innovative biotechnological processes (syngas fermentation to ethanol), and extractive/catalyzed processes for the valorization of cardoon roots to FDCA and residual solid biomass were modeled and simulated. Inulin conversion to 2,5-Furandicarboxylic acid was the main conversion route taken into consideration. Finally, the novel process flowsheet, treating 130,000 t/y of residual biomass and integrating all proposed technologies, was modeled and assessed using process simulation tools to achieve overall mass and energy balances for comparison with alternative options. The results indicated that cardoon biorefining through the proposed flowsheet can produce, per 1000 tons of input dry biomass, 211 kg of 2,5-Furandicarboxylic acid and 140 kg of ethanol through biomass gasification followed by syngas fermentation. Furthermore, a pre-feasibility analysis was conducted, revealing significant and potentially disruptive results in terms of environmental impact (with 40 ktCO2eq saved) and economic feasibility (with an annual gross profit of EUR 30 M/y).

Bioethanol can be produced from several biomasses including lignocellulosic materials. Besides 6-carbon sugars that represent the prevalent carbohydrates, some of these feedstocks contain significant amounts of 5-carbon sugars. One common limit of the major part of the xylose-fermenting yeasts is the diauxic shift between the uptake of glucose and xylose during the fermentation of mixed syrups. Thus, optimized fermentation strategies are required. In this paper the ability of Scheffersomyces stipitis strain NRRLY-11544 to ferment mixed syrups with a total sugar concentration in the range 40-80 g/L was investigated by using mono cultures, co-cultures with Saccharomyces cerevisiae strain Bakers Yeast Type II and single cultures immobilized in silica-hydrogel films. The experimental design for the fermentations with immobilized cells included the process analysis in function of two parameters: the fraction of the gel in the broth and the concentration of the cells loaded in the gel. Furthermore, for each total sugars level, the fermentative course of S. stipitis was analyzed at several glucose-to xylose ratios. The results indicated that the use of S. stipitis and S. cerevisiae in free co-cultures ensured faster processes than single cultures of S. stipitis either free or immobilized. However, the rapid production of ethanol by S. cerevisiae inhibited S. stipitis and caused a stuck of the process. Immobilization of S. stipitis in silica-hydrogel increased the relative consumption rate of xylose-to-glucose by 2-6 times depending on the composition of the fermentation medium. Furthermore the films performances appeared stable over three weeks of continuous operations. However, on the whole, the final process yields obtained with the immobilized cells were not meaningfully different from that of the free cells. This was probably due to concurrent fermentations operated by the cells released in the broth. Optimization of the carrier characteristics could improve the performances of the process with immobilized cells.

Industries currently rely on chicory and Jerusalem artichoke for inulin extraction but also cardoon is proved to synthetize and store high quantity of inulin in roots as well. Cardoon is a multipurpose crop, well adapted to marginal lands, whose main residues at the end of cropping cycle consist of roots. However, cardoon roots are a suitable source of inulin, that is of high interest for new generation biodegradable bioplastics production. On the other hand, a sustainable supply chain for inulin production from cardoon roots has not been developed yet. In particular, in the inulin supply chain the most critical part is storage, which can negatively affect both cost and inulin quantity. In the present study the effect on inulin content in cardoon roots stored as dried chipped roots (CRt) and dried whole roots (WRt) was investigated in a 6-month storage trial. Our findings suggest that chipping before storage did not affect the inulin content during the storage. Furthermore, it reduced the time needed for drying by 33.3% and increased the bulk density by 154.9% with the consequent reduction of direct cost for drying, transportation and storage.