Search Results (23)

The use of an ammonia fiber expansion pretreatment using low-moisture anhydrous ammonia (LMAA) is a promising strategy for biomass deconstruction, with significant effects on depolymerizing lignin and hemicellulose. An LMAA pretreatment provides several advantages, including compatibility with the high-biomass loading of solids, efficient ammonia recovery, and scalability for industrial operations. In this study, the reactor was revisited and optimized to improve glucan digestibility from corn stover through enzymatic hydrolysis, building on our previous findings that identified limitations in ammonia distribution. The effects of the biomass particle size, the reaction time, and their interaction on glucose yields were investigated to determine their influence on the subsequent enzymatic hydrolysis kinetics. The best glucose yield of 83% was achieved using an LMAA pretreatment of biomass with a 0.5 mm particle size, representing an improvement of approximately 5% compared to biomass with a 1 mm particle size. Additionally, reactor optimization led to a 22% improvement in the glucose yield compared to the previous reactor configuration. According to the results of the reaction kinetics fitting, the enzymatic hydrolysis data indicated that the reaction followed a pseudo-first-order model. Full article

In recent years, biofuels and bioenergy derived from algae have gained increasing attention, fueled by the growing demand for renewable energy sources and the urgent need to lower CO₂ emissions. This research examines the generation of bioethanol and biomethane using freshly harvested and sedimented algal biomass. Employing a factorial experimental design, various trials were conducted, with ethanol yield as the primary optimization target. The findings indicated that the sodium hydroxide concentration during pretreatment and the amylase dosage in enzymatic hydrolysis were key parameters influencing the ethanol production efficiency. Under optimized conditions—using 0.3 M NaOH, 25 μL/g starch, and 250 μL/g cellulose—fermentation yielded ethanol concentrations as high as 2.75 ± 0.18 g/L (45.13 ± 2.90%), underscoring the significance of both enzyme loading and alkali treatment. Biomethane potential tests on the residues of fermentation revealed reduced methane yields in comparison with the raw algal feedstock, with a peak value of 198.50 ± 25.57 mL/g volatile solids. The integrated process resulted in a total energy recovery of up to 809.58 kWh per tonne of algal biomass, with biomethane accounting for 87.16% of the total energy output. However, the energy recovered from unprocessed biomass alone was nearly double, indicating a trade-off between sequential valorization steps. A comparison between fresh and dried feedstocks also demonstrated marked differences, largely due to variations in moisture content and biomass composition. Overall, this study highlights the promise of integrated algal biomass utilization as a viable and energy-efficient route for sustainable biofuel production. Full article

This study examines the effect of surfactant-enhanced enzymatic hydrolysis on eucalyptus Kraft pulps produced under high (CPHA) and mild (CPMA) alkali conditions to optimize saccharification and sugar yield. Compositional analysis revealed an increase in glucan content, from 40% in untreated eucalyptus to 70.1% in CPHA. Both pulps were hydrolyzed using Cellic^® CTec3 HS enzyme (Novozymes). A 2² factorial design revealed maximum sugar conversion (~100%) with enzyme loading of 10 FPU/g carbohydrate and 10% (w/v) solids. Tween 20 significantly boosted hydrolysis in CPMA, increasing reducing sugars from 42 g/L to 65 g/L and efficiency from 59.6% to 92.2% within 6 h. By contrast, Tween 80 and PEG 400 showed limited effects on CPMA. Surfactants mitigated lignin–enzyme interactions, especially in CPMA, as higher lignin content restricted hydrolysis efficiency. Phenolic content in the hydrolysates revealed that Tween 80 increased the release of inhibitory compounds, while Tween 20 kept phenolic levels lower. Overall, Tween 20 improved sugar yields and hydrolysis efficiency even with moderate lignin removal during kraft pretreatment, highlighting its potential to reduce enzyme loading and costs in industrial biorefineries. This study underscores the importance of optimizing surfactant selection based on biomass composition for effective enzymatic hydrolysis for cellulosic sugar recovery. Full article

The bioconversion of lignocellulosic biomass, which are abundant and renewable resources, into liquid fuels and bulk chemicals is a promising solution to the current challenges of resource scarcity, energy crisis, and carbon emissions. Considering the separation of some end-products, it is necessary to firstly obtain a high concentration separated fermentable sugar solution, and then conduct fermentation. For this purpose, in this study, using acid catalyzed steam explosion pretreated corn stover (ACSE-CS) and corn cob residues (CCR) as cellulosic substrate, respectively, the batch feeding strategies and enzymatic hydrolysis conditions were investigated to achieve the efficient enzymatic hydrolysis at high solid loading. It was shown that the fermentable sugar solutions of 161.2 g/L and 205 g/L were obtained, respectively, by fed-batch enzymatic hydrolysis of ACSE-CS under 30% of final solid loading with 10 FPU/g DM of crude cellulase, and of CCR at 27% of final solid loading with 8 FPU/g DM of crude cellulase, which have the potential to be directly applied to the large-scale fermentation process without the need for concentration, and the conversion of glucan in ACSE-CS and CCR reached 80.9% and 87.6%, respectively, at 72 h of enzymatic hydrolysis. This study also applied the fed-batch simultaneous saccharification and co-fermentation process to effectively convert the two cellulosic substrates into ethanol, and the ethanol concentrations in fermentation broth reached 46.1 g/L and 72.8 g/L for ACSE-CS and CCR, respectively, at 144 h of fermentation. This study provides a valuable reference for the establishment of “sugar platform” based on lignocellulosic biomass and the production of cellulosic ethanol. Full article

The study involves the use of commercial cellulase Cellic CTec2 in combination with two in-house xylanases, PxXyn10A (XynA), a recombinant purified enzyme from Paenibacillus xylanivorans A59, and a xylanase enzymatic extract from native Moesziomyces aphidis PYCC 5535T (MaPYCC 5535T), for the enzymatic hydrolysis of pretreated blue agave bagasse (BAB) at the high solids load of 20% (w/v). Three different combinations of cellulase and xylanases were evaluated. When Cellic^® CTec2 was used at a dosage of 10 FPU/g oven-dried solids (ODS) supplemented with XynA or MaPYCC 5535T at an endo-xylanase dosage of 100 U/g ODS, increases in the xylose yield of 30% and 33%, respectively, were obtained. When applying in-house xylanases alone (at an endo-xylanase dosage of 100 U/g ODS), xylan in BAB was selectively hydrolyzed into xylose with 5% yield with MaPYCC 5535T, while no xylose was detected with XynA. Interestingly, a synergic effect of Cellic^® CTec 2 with both xylanases was observed when using a low dosage of 1 FPU/g ODS (allowing for some liquefaction of the reaction mixture), promoting xylose and glucose release by either xylanase. A higher concentration of monomeric sugars was obtained with 10 FPU/g ODS of Cellic^® Ctec 2 supplemented with 100 U/g ODS of MaPYCC 5535T, followed by XynA. The improvement in saccharification through the synergistic combination of in-house xylanases and commercial cellulases allows for the obtention of sugar-rich hydrolysates, which enhances the technical sustainability of the process. Hydrolysates were then fermented using recombinant Cellux 4^TM yeast to yield 45 g/L ethanol, representing an increase of about 30% with respect to the control obtained with only the commercial cellulase cocktail. The surface modification of agave biomass with the different combinations of enzymes was evidenced by scanning electron microscopy (SEM). Full article

Cellulosic ethanol is the key technology to alleviate the pressure of energy supply and climate change. However, the ethanol production process, which is close to industrial production and has a high saccharification rate and ethanol yield, still needs to be developed. This study demonstrates the effective conversion of poplar wood waste into fuel-grade ethanol. By employing a two-step pretreatment using sodium chlorite (SC)-dilute sulfuric acid (DSA), the raw material achieved a sugar conversion rate exceeding 85% of the theoretical value. Under optimized conditions, brewing yeast co-utilizing C6/C5 enabled a yield of 35 g/L ethanol from 10% solid loading delignified poplar hydrolysate. We increased the solid loading to enhance the final ethanol concentration and optimized both the hydrolysis and fermentation stages. With 20% solid loading delignified poplar hydrolysate, the final ethanol concentration reached 60 g/L, a 71.4% increase from the 10% solid loading. Our work incorporates the pretreatment, enzymatic hydrolysis, and fermentation stages to establish a simple, crude poplar waste fuel ethanol process, expanding the range of feedstocks for second-generation fuel ethanol production. Full article

Coffee silverskin (CS), a by-product of the coffee roasting process, has high protein content (16.2−19.0%, w/w), making it a potential source for plant protein and bioactive peptide production. This study aims to develop innovative extraction methods for phenolic compounds and proteins from CS. The conditions for hydrothermal (HT) extraction of phenolic compounds from CS were optimized by varying CS loading (2.5−10%, w/v), temperature (110−130 °C), and time (5−30 min) using a one-factor-at-a-time (OFAT) approach. The highest TPC of 55.59 ± 0.12 µmole GAE/g _CS was achieved at 5.0% (w/v) CS loading and autoclaving at 125 °C for 25 min. Following hydrothermal extraction, CS protein was extracted from HT-extracted solid fraction by microwave-assisted alkaline extraction (MAE) using 0.2 M NaOH at 90 W for 2 min, resulting in a protein recovery of 12.19 ± 0.39 mg/g _CS. The CS protein was then subjected to enzymatic hydrolysis using protease from Bacillus halodurans SE5 (protease_SE5). Protease_SE5-derived CS protein hydrolysate had a peptide concentration of 0.73 ± 0.09 mg/mL, with ABTS, DPPH, and FRAP values of 15.71 ± 0.10, 16.63 ± 0.061, and 6.48 ± 0.01 µmole TE/mL, respectively. Peptide identification by LC-MS/MS revealed several promising biological activities without toxicity or allergenicity concerns. This study’s integrated approach offers a sustainable and efficient method for extracting valuable compounds from CS, with potential applications in the food and pharmaceutical industries. Full article

Extracted sugar beet press pulp (SBPP) is a promising agricultural residue for saccharification and further bioconversion. Combining solid-state fermentation of SBPP with engineered Aspergillus niger for enzyme production followed by hydrolysis of additionally added SBPP in the same bioreactor was studied to produce a sugar solution (hydrolysate) in a one-pot process. The initial aerobic solid-state fermentations were carried out in duplicate on non-milled, wet SBPP (moisture content of 72% (w/v)) with an A. niger strain engineered for constitutive pectinase production for 96 h, and this resulted in polygalacturonase activities of up to 256 U mL⁻¹ in the wet media. Afterwards, water was added to the bioreactor, and the remaining solids were suspended by stirring to dissolve the hydrolytic enzymes. Metabolic activities of A. niger were inactivated by a N₂-atmosphere and by increasing the temperature to 50 °C. High solid loads of milled SBPP were added to the stirred-tank reactor with a delay of 24 h to enable sugar yield calculations based on the compositional analysis of the SBPP used. The resulting final sugar concentrations of the hydrolysate after 166 h were 17 g L⁻¹ d-glucose, 18.8 g L⁻¹ l-arabinose, and 12.5 g L⁻¹ d-galacturonic acid, corresponding to sugar yields of 98% d-glucose, 86% l-arabinose, and 50% d-galacturonic acid, respectively. Including the other sugars released during enzymatic hydrolysis in the one-pot process (d-xylose, d-mannose, d-galactose), a total sugar concentration of 54.8 g L⁻¹ was achieved in the hydrolysate. The one-pot process combining hydrolytic enzyme production in solid-state fermentation with high solid loads during enzymatic hydrolysis of the milled SBPP reduces hydrolytic process costs by replacing chemical pre-treatments, enabling the in situ production of SBPP-adapted hydrolytic enzymes, as well as avoiding intermediate enzyme extraction and preparation steps. Full article

Brewer’s spent grain (BSG) as the major byproduct in the brewing industry is a promising feedstock to produce value-added products such as volatile fatty acids (VFAs). Synergistic ball mill–enzymatic hydrolysis (BM-EH) process is an environmentally friendly pretreatment method for lignocellulosic materials before bioprocessing. This study investigated the potential of raw and BM-EH pretreated BSG feedstocks to produce VFAs through a direct thermophilic anaerobic fermentation process without introducing a methanogen inhibitor. The highest VFA concentration of over 30 g/L was achieved under the high-solid loading fermentation (HS) of raw BSG. The synergistic BM-EH pretreatment helps to increase the cellulose conversion to 70%. Under conventional low TS fermentation conditions, compared to the controlled sample, prolonged pretreatment of the BSG substrate resulted in increased VFA yields from 0.25 to 0.33 g/g_VS, and butyric acid became dominant instead of acetic acid. Full article

Cellulosic sugars production for the valorization of lignocellulosic biomass residues in an industrial site has economic benefits and is promising if integrated into a biorefinery. Enzymatic hydrolysis (EH) of pretreated Eucalyptus globulus bark, an industrial residue of low-economic value widely available in Portuguese pulp and paper mills, could be an excellent approach to attain resource circularity and pulp mill profitability. This work evaluated the potential for improving cellulosic sugars concentrations by operating with high solids loading and introducing the additives Triton X-100, PEG 4000 and Tween 80 using a commercial enzymatic consortium with a dosage of 25 FPU g_{carbohydrates}⁻¹. Additives did not improve enzymatic hydrolysis performance, but the effect of increasing solids loading to 14% (w/v) in batch operation was accomplished. The fed-batch operation strategy was investigated and, when starting with 11% (w/v) solids loading, allowed the feeding of 3% (w/v) fresh feedstock sequentially at 2, 4 and 6 h, attaining 20% (w/v) total solids loading. After 24 h of operation, the concentration of cellulosic sugars reached 161 g L⁻¹, corresponding to an EH conversion efficiency of 76%. Finally, the fermentability of the fed-batch hydrolysate using the Ethanol Red^® strain was evaluated in a 5 L bioreactor scale. The present results demonstrate that Eucalyptus globulus bark, previously pretreated by kraft pulping, is a promising feedstock for cellulosic sugars production, allowing it to become the raw material for feeding a wide range of bioprocesses. Full article

Lignocellulosic biomass (LCB) has remained a latent alternative resource to be the main substitute for oil and its derivatives in a biorefinery concept. However, its complex structure and the underdeveloped technologies for its large-scale processing keep it in a state of constant study trying to establish a consolidated process. In intensive processes, enzymes have been shown to be important molecules for the fractionation and conversion of LCB into biofuels and high-value-added molecules. However, operational challenges must be overcome before enzyme technology can be the main resource for obtaining second-generation sugars. The use of additives is shown to be a suitable strategy to improve the saccharification process. This review describes the mechanisms, roles, and effects of using additives, such as surfactants, biosurfactants, and non-catalytic proteins, separately and integrated into the enzymatic hydrolysis process of lignocellulosic biomass. In doing so, it provides a technical background in which operational biomass processing hurdles such as solids and enzymatic loadings, pretreatment burdens, and the unproductive adsorption phenomenon can be addressed. Full article

Corncob is a potential feedstock for biorefineries to produce cellulosic ethanol and other chemicals. Densifying lignocellulosic biomass with chemicals followed by autoclave (DLCA) has been confirmed an efficient and economical pretreatment method, and it was applied in the present work for conversion of corncob to bioethanol. The dosage of sulfuric acid, solid loading of biomass, and autoclave time for pretreatment were investigated. Enzymatic hydrolysis at 25–35% solids loadings resulted in 91–97% sugar conversions. Fermentation of the resulted hydrolysates went well with the highest ethanol titer reaching 75.71 g/L at 35% solid loading. Simultaneous saccharification and co-fermentation was applied to further improve xylose consumption at high solids loadings and the ethanol titer was enhanced to 82.0 g/L at 35% solid loading with an ethanol yield of 21.67 kg/100 kg corncob. This study demonstrated DLCA provided a highly digestible and highly fermentable corncob for biorefinery. Full article

Being a lignocellulose-rich biomass, pineapple leaves waste (PL) could be a potential raw material for the production of biofuel, biochemicals, and other value-added products. The main aim of this study was to investigate the potential of pineapple leaves in the sustainable production of bioethanol via stepwise saccharification and fermentation. For this purpose, PL was subjected to hydrothermal pretreatment in a high-pressure reactor at 150 °C for 20 min without any catalyst, resulting in a maximum reducing sugar yield of 38.1 g/L in the liquid fraction after solid-liquid separation of the pretreated hydrolysate. Inhibitors (phenolics, furans) and oligomers production were also monitored during the pretreatment in the liquid fraction of pretreated PL. Enzymatic hydrolysis (EH) of both pretreated biomass slurry and cellulose-rich solid fraction maintained at a solid loading (dry basis) of 5% wt. was performed at 50 °C and 150 rpm using commercial cellulase at an enzyme dose of 10 FPU/gds. EH resulted in a glucose yield of 13.7 and 18.4 g/L from pretreated slurry and solid fractions, respectively. Fermentation of the sugar syrup obtained by EH of pretreated slurry and the solid fraction was performed at 30 °C for 72 h using Saccharomyces cerevisiae WLP300, resulting in significant ethanol production with more than 91% fermentation efficiency. This study reveals the potential of pineapple leaves waste for biorefinery application, and the role of inhibitors in the overall efficiency of the process when using whole biomass slurry as a substrate. Full article

Incorporation of various alternative resources as co-digestion substrates aids to reduce the consumption of agricultural crops for biogas production. However, the efficiency and limitations of these co-substrates is still not fully understood. Use of biomass waste remaining after enzymatic hydrolysis for high value chemical fermentation, meat processing and dairy wastewater primary sludge as co-substrates in an agricultural resource anaerobic digestion plant is tackled within this study. The results showed that anionic surfactants (<200 ppm) can be used to improve fat, oil and grease (FOG) solubility in water and, at the same time, enhance the biomethane potential of FOG-containing sludge by increasing it from 1374.5 to 1765 mLCH₄/gVS for meat processing wastewater primary sludge, and from 534 to 740 mLCH₄/gVS for dairy wastewater primary sludge, when agricultural digestate is used as a substrate and sludge loading is not more than 10% from the volatile solids loaded. At the same time, only 549.7 mLCH₄/gVS was produced as 30-day BMP when 5% biomass hydrolysis waste was used. Biomass hydrolysis waste co-digestion with primary sludge from dairy and meat processing wastewaters has an antigenic effect, and separate substrate anaerobic digestion gave a better results, thus, showing that excessive combination of various waste resources can be inhibitory for biogas production and the appropriate substrate selection and combination is a technical challenge for the biogas industry. Full article

Growth in population and thereby increased industrialization to meet its requirement, has elevated significantly the demand for energy resources. Depletion of fossil fuel and environmental sustainability issues encouraged the exploration of alternative renewable eco-friendly fuel resources. Among major alternative fuels, bio-ethanol produced from lignocellulosic biomass is the most popular one. Lignocellulosic biomass is the most abundant renewable resource which is ubiquitous on our planet. All the plant biomass is lignocellulosic which is composed of cellulose, hemicellulose and lignin, intricately linked to each other. Filamentous fungi are known to secrete a plethora of biomass hydrolyzing enzymes. Mostly these enzymes are inducible, hence the fungi secrete them economically which causes challenges in their hyperproduction. Biomass’s complicated structure also throws challenges for which pre-treatments of biomass are necessary to make the biomass amorphous to be accessible for the enzymes to act on it. The enzymatic hydrolysis of biomass is the most sustainable way for fermentable sugar generation to convert into ethanol. To have sufficient ethanol concentration in the broth for efficient distillation, high solid loading >20% of biomass is desirable and is the crux of the whole technology. High solid loading offers several benefits including a high concentration of sugars in broth, low equipment sizing, saving cost on infrastructure, etc. Along with the benefits, several challenges also emerged simultaneously, like issues of mass transfer, low reaction rate due to water constrains in, high inhibitor concentration, non-productive binding of enzyme lignin, etc. This article will give an insight into the challenges for cellulase action on cellulosic biomass at a high solid loading of biomass and its probable solutions. Full article