Production of biohydrogen from sugars and lignocellulosic biomass using Thermoanaerobacter GHL15

The effect of culture parameters on hydrogen production using strain GHL₁₅ in batch culture was investigated. The strain belongs to the genus Thermoanaerobacter with 98.9% similarity to Thermoanaerobacter yonseiensis and 98.5% to Thermoanaerobacter keratinophilus with a temperature optimum of 65–70 °C and a pH optimum of 6–7. The strain metabolizes various pentoses, hexoses, and disaccharides to acetate, ethanol, hydrogen, and carbon dioxide. However substrate inhibition was observed above 10 mM glucose concentration. Maximum hydrogen yields on glucose were 3.1 mol H₂ mol⁻¹ glucose at very low partial pressure of hydrogen. Hydrogen production from various lignocellulosic biomass hydrolysates was investigated in batch culture. Various pretreatment methods were examined including acid, base, and enzymatic (Celluclast^® and Novozyme 188) hydrolysis. Maximum hydrogen production (5.8–6.0 mmol H₂ g⁻¹ dw) was observed from Whatman paper (cellulose) hydrolysates although less hydrogen was produced by hydrolysates from other examined lignocellulosic materials (maximally 4.83 mmol H₂ g⁻¹ dw of grass hydrolysate). The hydrogen yields from all lignocellulosic hydrolysates were improved by acid and alkaline pretreatments, with maximum yields on grass, 7.6 mmol H₂ g⁻¹ dw.     

Comparative study on ethanol production from pretreated sugarcane bagasse using immobilized Saccharomyces cerevisiae on various matrices

Microwave alkali pretreated sugarcane bagasse was used as a substrate for production of cellulolytic enzymes, needed for biomass hydrolysis. The pretreated sugarcane bagasse was enzymatic hydrolyzed by crude unprocessed enzymes cellulase (Filter paper activity 9.4 FPU/g), endoglucanase (carboxymethylcellulase, 148 IU/g), β-glucosidase (116 IU/g) and xylanase (201 IU/g) produced by Aspergillus flavus using pretreated sugarcane bagasse as substrate under solid state fermentation. Concentrated enzymatic hydrolyzate was used for ethanol production using Saccharomyces cerevisiae immobilized on various matrices. The yield of ethanol was 0.44 g_p/g_s in case of yeast immobilized sugarcane bagasse, 0.38 g_p/g_s using Ca-alginate and 0.33 g_p/g_s using agar-agar as immobilization matrices. The immobilized yeast studied up to 10 cycles in case of immobilized sugarcane bagasse and up to 4 cycles in case of agar-agar and calcium alginate for ethanol production under repeated batch fermentation study.     

Ethanol production by Mucor indicus and Rhizopus oryzae from rice straw by separate hydrolysis and fermentation

Rice straw was successfully converted to ethanol by separate enzymatic hydrolysis and fermentation by Mucor indicus, Rhizopus oryzae, and Saccharomyces cerevisiae. The hydrolysis temperature and pH of commercial cellulase and β-glucosidase enzymes were first investigated and their best performance obtained at 45 °C and pH 5.0. The pretreatment of the straw with dilute-acid hydrolysis resulted in 0.72 g g^?1 sugar yield during 48 h enzymatic hydrolysis, which was higher than steam-pretreated (0.60 g g^?1) and untreated straw (0.46 g g^?1). Furthermore, increasing the concentration of the dilute-acid pretreated straw from 20 to 50 and 100 g L^?1 resulted in 13% and 16% lower sugar yield, respectively. Anaerobic cultivation of the hydrolyzates with M. indicus resulted in 0.36–0.43 g g^?1 ethanol, 0.11–0.17 g g^?1 biomass, and 0.04–0.06 g g^?1 glycerol, which is comparable with the corresponding yields by S. cerevisiae (0.37–0.45 g g^?1 ethanol, 0.04–0.10 g g^?1 biomass and 0.05–0.07 glycerol). These two fungi produced no other major metabolite from the straw and completed the cultivation in less than 25 h. However, R. oryzae produced lactic acid as the major by-product with yield of 0.05–0.09 g g^?1. This fungus had ethanol, biomass and glycerol yields of 0.33–0.41, 0.06–0.12, and 0.03–0.04 g g^?1, respectively.     

Ethanol and biogas production from birch by NMMO pretreatment

Birch wood was pretreated with N-methylmorpholine-N-oxide (NMMO or NMO) followed by enzymatic hydrolysis and fermentation to ethanol or digestion to biogas. The pretreatments were carried out with NMMO (w_NMMO = 85%) at 130 °C for 3 h, and the effects of drying after the pretreatment were investigated. Enzymatic hydrolysis of the untreated wood resulted in 8%–10% of theoretical glucose yield after 4 days hydrolysis, while the NMMO pretreatment improved this yield to 91%. Consequently, ethanol production yield from NMMO-pretreated materials resulted in around 9-fold improvement compared to the untreated wood. On the other hand, drying of the pretreated wood had a negative impact and decreased the yield of enzymatic hydrolysis by 4%–10%. Digestion of the untreated wood with thermophilic bacteria resulted in maximum methane yield of 158 cm³ g⁻¹ of VS in 30 days, while the NMMO pretreatment improved the methane yield up to 232 cm³ g⁻¹ of VS (80% of the theoretical biogas yield) in just 9 days.     

Lipid production from sweet sorghum bagasse through yeast fermentation

Cryptococcus curvatus has great potential in fermenting unconditioned hydrolysates of sweet sorghum bagasse. With hydrolysates obtained by enzymatic hydrolysis of the solid pretreated by microwave with lime, the maximal yeast cell dry weight and lipid content were 10.83 g/l and 73.26%, respectively. For hydrolysates obtained in the same way but without lime, these two parameters were 15.50 g/l and 63.98%, respectively. During yeast fermentation, glucose and xylose were consumed simultaneously while cellobiose was released from the residual bagasse. The presence of lime, on one hand, made cellulose more accessible to enzymes as evidenced by higher total reducing sugar release compared to that without during enzymatic hydrolysis step; on the other hand, it caused the degradation of sugars to non-sugar chemicals during pretreatment step. As a result, higher lipid yield of 0.11 g/g bagasse or 0.65 ton/hectare of land was achieved from the pathway of microwave pretreatment and enzymatic hydrolysis while 0.09 g/g bagasse or 0.51 ton/hectare of land was attained from the process of lime-assisted microwave pretreatment followed by the same enzymatic saccharification.     

Empty fruit bunches from oil palm as a potential raw material for fuel ethanol production

In this paper the fuel ethanol production from empty fruit bunches was experimentally evaluated using alkaline pretreatment and enzymatic hydrolysis for sugars release. Fermentation was accomplished using a native Saccharomyces cerevisiae strain. Ethanol concentration was carried on using a glass bench-scale distillation column. Experimental results were used for planning and designing the process scheme using Aspen Plus. Process simulation allowed calculating the mass and energy balances. It was found that coupling alkaline pretreatment with a later autoclaving improved the sugars yield in enzymatic hydrolysis. However, the use of the remaining soaking solution from pretreatment as hydrolysis medium had negative effects on sugars yield suggesting that there exist inhibit substance for the enzyme. Better results for enzymatic hydrolysis were obtained when sodium acetate buffer was used. Ethanol yield obtained from both experiments and simulation were very similar (66.50 and 65.84 dm³ of ethanol per each t of empty fruit bunches, respectively). These low ethanol yields were obtained because the native S. cerevisiae does not assimilate all reducing sugars, suggesting that those sugars were pentoses. Simulated alkaline and autoclaving pretreatment contributed only with 2% of the total energy consumption (198.4 GJ m⁻³ ethanol) while product recovery represented 57% of the total energy.     

Enhanced enzymatic conversion with freeze pretreatment of rice straw

Production of bioethanol by the conversion of lignocellulosic waste has attracted much interest in recent years, because of its low cost and great potential availability. The pretreatment process is important for increasing the enzymatic digestibility of lignocellulosic materials. Enzymatic conversion with freeze pretreatment of rice straw was evaluated in this study. The freeze pretreatment was found to significantly increase the enzyme digestibility of rice straw from 48% to 84%. According to the results, enzymatic hydrolysis of unpretreated rice straw with 150 U cellulase and 100 U xylanase for 48 h yielded 226.77 g kg⁻¹ and 93.84 g kg⁻¹ substrate-reducing sugars respectively. However, the reducing sugar yields from freeze pretreatment under the same conditions were 417.27 g kg⁻¹ and 138.77 g kg⁻¹ substrate, respectively. In addition, hydrolyzates analysis showed that the highest glucose yield obtained during the enzymatic hydrolysis step in the present study was 371.91 g kg⁻¹ of dry rice straw, following pretreatment. Therefore, the enhanced enzymatic conversion with freeze pretreatment of rice straw was observed in this study. This indicated that freeze pretreatment was highly effective for enzymatic hydrolysis and low environmental impact.     

Comparison of four different chemical pretreatments of corn stover for enhancing enzymatic digestibility

Four commonly used chemical pretreatment processes based on dilute acid, lime, aqueous ammonia steeping followed by dilute acid hydrolysis, and sodium hydroxide, were evaluated to provide comparative performance data. An obverse correlation between lignin removal and enzymatic digestibility of pretreated corn stover was observed. Compared with other three pretreatments, pretreatment of corn stover with 2% NaOH substantially increased the lignin removal and enhanced the accessibility and digestibility of cellulose. The hydrolysis yield of NaOH-pretreated corn stover reached 81.2% by 48 h at 8.0% substrate concentration and cellulase dosage of 20 FPU g⁻¹ substrate. Chemical analysis showed that the enzymatic hydrolysate from NaOH-pretreated corn stover contained higher content of fermentable sugars and less inhibitors, which is suitable for subsequent fermentation process to produce ethanol. The research results are meaningful in bioconversion and utilization of renewable lignocellulosic biomass.     

Conversion of paper sludge to ethanol by separate hydrolysis and fermentation (SHF) using Saccharomyces cerevisiae

The conversion of ethanol from paper sludge using the separate hydrolysis and fermentation (SHF) process with cellulase and Saccharomyces cerevisiae GIM-2 were investigated in this paper. Optimization strategy based on statistical experimental designs was employed to enhance degree of saccharification by enzymatic hydrolysis of paper sludge. Based on the Plackett-Burman design, hydrolysis time, substrate concentration and cellulase dosage were selected as the most significant variable on the degree of saccharification. Subsequently, the optimum combination of the selected factors was investigated by a Box-Behnken approach. A mathematical model was developed to show the effects of each factor and their combinatorial interactions on the degree of saccharification. The optimal conditions were hydrolysis time 82.7 h, substrate concentration 40.8 g L⁻¹ and cellulase dosage 18.1 FPU g⁻¹ substrate, and a degree of saccharification of 82.1% can be achieved. When hydrolysate was further fermented with S. cerevisiae GIM-2, the conversion rate of sugar to ethanol was 34.2% and the ethanol yield was 190 g kg⁻¹ of dry paper sludge, corresponding to an overall conversion yield of 56.3% of the available carbohydrates on the initial substrate. The results derived from this study indicate that the response surface methodology is a useful tool for optimizing the hydrolysis conditions to converse paper sludge to ethanol.     

Utilization of pineapple stem juice to enhance enzyme-hydrolytic efficiency for sugarcane bagasse after an optimized pre-treatment with alkaline peroxide

The enzymatic hydrolysis of sugarcane bagasse was investigated by treating a peroxide–alkaline bagasse with a pineapple stem juice, xylanase and cellulase. Pre-treatment procedures of sugarcane bagasse with alkaline hydrogen peroxide were evaluated and compared. Analyses were performed using 2⁴ factorial designs, with pre-treatment time, temperature, magnesium sulfate and hydrogen peroxide concentration as factors. The responses evaluated were the yield of cellobiose and glucose released from pretreated bagasse after enzymatic hydrolysis. The results show that the highest enzymatic conversion was obtained for bagasse using 2% hydrogen peroxide at 60 °C for 16 h in the presence of 0.5% magnesium sulfate. Bagasse (5%) was treated with pineapple stem extract, which contains mixtures of protease and esterase, in combination with xylanase and cellulase. It was observed that the amount of glucose and cellobiose released from bagasse increased with the mixture of enzymes. It is believed that the enzymes present in pineapple extracts are capable of hydrolyze specific linkages that would facilitate the action of digesting plant cell walls enzymes. This increases the amount of glucose and other hexoses that are released during the enzymatic treatment and also reduces the amount of cellulase necessary in a typical hydrolysis.     

Cellulase production using biomass feed stock and its application in lignocellulose saccharification for bio-ethanol production

A major constraint in the enzymatic saccharification of biomass for ethanol production is the cost of cellulase enzymes. Production cost of cellulases may be brought down by multifaceted approaches which include the use of cheap lignocellulosic substrates for fermentation production of the enzyme, and the use of cost efficient fermentation strategies like solid state fermentation (SSF). In the present study, cellulolytic enzymes for biomass hydrolysis were produced using solid state fermentation on wheat bran as substrate. Crude cellulase and a relatively glucose tolerant BGL were produced using fungi Trichoderma reesei RUT C30 and Aspergillus niger MTCC 7956, respectively. Saccharification of three different feed stock, i.e. sugar cane bagasse, rice straw and water hyacinth biomass was studied using the enzymes. Saccharification was performed with 50 FPU of cellulase and 10 U of β-glucosidase per gram of pretreated biomass. Highest yield of reducing sugars (26.3 g/L) was obtained from rice straw followed by sugar cane bagasse (17.79 g/L). The enzymatic hydrolysate of rice straw was used as substrate for ethanol production by Saccharomyces cerevisiae. The yield of ethanol was 0.093 g per gram of pretreated rice straw.     

Waste biomass to liquids: Low temperature conversion of sugarcane bagasse to bio-oil. The effect of combined hydrolysis treatments

This article describes the influence of different sugarcane bagasse hydrolysis pretreatments on modifications to biomass feedstock and the characteristics of the resultant pyrolysis products. Sugarcane bagasse was pretreated with acid, alkaline or sequential acid/alkaline solutions and pretreated samples were then subjected to a low temperature conversion (LTC) process under He or O₂/He atmospheres at 350-450 °C. Both pretreated samples and sugarcane bagasse in natura were analyzed by determination of their chemical composition and by thermogravimetric, FTIR and SEM analyses. The gases yielded during LTC were monitored on-line by quadrupole mass spectrometry, and the liquid fractions obtained were characterized by FTIR and ¹H and ¹³C NMR. Irrespective of the sugarcane bagasse pretreatment applied, the main bio-oil component obtained was levoglucosan. However, the LTC yield of bio-oil depended on the hydrolysis treatment of the biomass and decreased in the presence of O₂. The acid hydrolysis pretreatment increased the LTC bio-oil yield notably.     

Preliminary exploration on pretreatment with metal chlorides and enzymatic hydrolysis of bagasse

Converting biomass to fermentable sugar is the critical step in the biomass refinery. Moreover, pretreatment of biomass plays an important role in improving the conversion of biomass to sugar. In this study, sugarcane bagasse was pretreated by metal chloride Lewis acids (0.1 mol L⁻³ CrCl₃, FeCl₃, FeCl₂, ZnCl₂ and AlCl₃ solution) for cellulase hydrolysis. The effects of pretreatments on the yield, chemical components, and sequential cellulase hydrolysis of pretreated bagasse were investigated. The results indicated that metal chlorides with different pKa values could efficiently remove the hemicellulose in bagasse during pretreatment. Furthermore, an inhibition factor (IF) quantitatively reflecting difficulty of cellulase hydrolysis was proposed. The low IF means the facile cellulase hydrolysis. The IF of Fe (III)-pretreated bagasse could decrease to 1.35. In this case, the enzymatic digestibility of bagasse approached to 100%.     

Influence of impregnation with lactic acid on sugar yields from steam pretreatment of sugarcane bagasse and spruce, for bioethanol production

Lignocellulosic biomass can be utilized to produce ethanol, a promising alternative energy source produced through fermentation of sugars. However, in order to achieve high sugar and ethanol yields, the lignocellulosic material must be pretreated before the enzymatic hydrolysis and fermentation. Dilute acid pretreatment, using SO₂, is one of the most promising methods of pretreatment for softwood and agricultural residues. However, handling the high acidity of the slurry obtained from pretreatment and difficulty in recycling/degradation of the impregnating agent are some of the drawbacks of the dilute acid processes. In the present study the influence of utilization of a weak organic acid (lactic acid), as impregnating agent, on the sugar yield from pretreatment, with and without addition of SO₂, was investigated. The efficiency of pretreatment was assessed by enzymatic hydrolysis of the slurry obtained by pretreatment, using sugarcane bagasse and spruce, stored for one and two months in the presence of lactic acid separately, as feedstocks. Pretreatment of bagasse after storage with 0.5% lactic acid resulted in an overall glucose yield, i.e. after enzymatic hydrolysis, of 79% of theoretical based on the amount available in the raw material. This was as good as pretreatment using SO₂ as impregnating agent. However, storage of spruce with lactic acid before pretreatment, with and without addition of SO₂, was not efficient and resulted in lower sugar yields than pretreatment using SO₂ only.     

Comparison between heterofermentation and autofermentation in hydrogen production from Arthrospira (Spirulina) platensis wet biomass

Hydrogen production from Arthrospira (Spirulina) platensis wet biomass through heterofermentation by the [FeFe] hydrogenase of hydrogenogens (hydrogen-producing bacteria) and autofermentation by the [NiFe] hydrogenase of Arthrospira platensis was discussed under dark anaerobic conditions. In heterofermentation, wet cyanobacterial biomass without pretreatment was hardly utilized by hydrogenogens for hydrogen production. But the carbohydrates in cyanobacterial cells released after cell wall disruption were effectively utilized by hydrogenogens for hydrogen production. Wet cyanobacterial biomass was pretreated with boiling and bead milling, ultrasonication, and ultrasonication and enzymatic hydrolysis. Wet cyanobacterial biomass pretreated with ultrasonication and enzymatic hydrolysis achieved the maximum reducing sugar yield of 0.407 g/g-DW (83.0% of the theoretical reducing sugar yield). Different concentrations (10 g/l to 40 g/l) of pretreated wet cyanobacterial biomass were used as substrate to produce fermentative hydrogen by hydrogenogens, which were domesticated with the pretreated wet cyanobacterial biomass as carbon source. The maximum hydrogen yield of 92.0 ml H₂/g-DW was obtained at 20 g/l of wet cyanobacterial biomass. The main soluble metabolite products (SMPs) in the residual solutions from heterofermentation were acetate and butyrate. In autofermentation, hydrogen yield decreased from 51.4 ml H₂/g-DW to 11.0 ml H₂/g-DW with increasing substrate concentration from 1 g/l to 20 g/l. The main SMPs in the residual solutions from autofermentation were acetate and ethanol. The hydrogen production peak rate and hydrogen yield at 20 g/l of wet cyanobacterial biomass in heterofermentation showed 110- and 8.4-fold increases, respectively, relative to those in autofementation.     

Bioethanol production from sweet potato by co-immobilization of saccharolytic molds and Saccharomyces cerevisiae

To investigate the bioethanol production from sweet potato, the saccharification and fermentation conditions of co-immobilization of saccharolytic molds (Aspergillus oryzae and Monascus purpureus) with Saccharomyces cerevisiae were analyzed. The immobilized yeast cells showed that at 10% glucose YPD (yeast extract peptone dextrose) the maximum fermentation rate was 80.23%. Viability of yeasts cells were 95.70% at a final ethanol concentration of 6%. Immobilization enhanced the ethanol tolerance of yeast cells. In co-immobilization of S. cerevisiae with A. oryzae or M. purpureus, the optimal hardening time of gel beads was between 15 and 60 min. Bioethanol production was 3.05-3.17% (v v⁻¹) and the Y_E/s (yield of ethanol production/starch consumption) was 0.31-0.37 at pH 4, 30 °C and 150 rpm during 13 days fermentation period. Co-immobilization of S. cerevisiae with a mixed cultures of A. oryzae and M. purpureus at a ratio of 2:1, the bioethanol production was 3.84% (v v⁻¹), and the Y_E/s was 0.39 for a 11 days incubation. However a ratio of A. oryzae and M. purpureus at 1:2 resulted a bioethanol production rate of 4.08% (v v⁻¹), and a Y_E/s of 0.41 after 9 days of fermentation.     

Biohydrogen production from algal biomass (Anabaena sp. PCC 7120) cultivated in airlift photobioreactor

The present study deals with the optimization of pretreatment conditions followed by thermophilic dark fermentative hydrogen production using Anabaena PCC 7120 as substrate by mixed microflora. Different airlift photobioreactors with ratio of area of downcomer and riser (A_d/A_r) in range of 0.4–3.2 were considered. Maximum biomass concentration of 1.63 g L⁻¹ in 9 d under light intensity of 120 μE m⁻² s⁻¹ was observed at A_d/A_r of 1.6. The mixing time of the reactors was inversely proportional to A_d/A_r. Maximal H₂ production was found to be 1600 mL L⁻¹ upon pretreatment with amylase followed by thermophilic fermentation for 24 h compared to other methods like sonication (200 mL L⁻¹), autoclave (600 mL L⁻¹) and HCl treatment (1230 mL L⁻¹). The decrease of pH from 6.5 to 5.0 during fermentation was due to the accumulation of volatile fatty acids. Amylase pretreatment gave higher reducible sugar content of 7.6 g L⁻¹ as compare to other pretreatments. Thermophilic fermentation of pretreated Anabaena biomass by mixed bacterial culture was found suitable for H₂ production.     

Preliminary study on enzymatic hydrolysis of treated oil palm (Elaeis) empty fruit bunches fibre (EFB) by using combination of cellulase and �� 1-4 glucosidase

Preliminary study on enzymatic hydrolysis process using combination of cellulase and ?? 1-4 glucosidase on treated oil palm empty fruit bunch fibre (EFB) was performed. Crucial trends for parameters such as pH, temperature and substrate loading influencing the enzymatic hydrolysis of the treated EFB fibre were also studied. Results revealed that a combination of both cellulase and ?? 1-4 glucosidase with the ratio of 5:1 hydrolyzed more cellulose from treated EFB fibre and gave highest soluble glucose concentration up to 4 g L⁻¹. The results indicated that as pH and temperature were increased the glucose produced also increased until pH 4.8 and 50 °C; beyond these values the reverse occurred. Glucose produced in the reaction increased with the increment in the substrate loading and maximum glucose concentration (2.7 g L⁻¹) was achieved when 8% (wv⁻¹) treated EFB was used as a substrate.     

Studying the ability of Fusarium oxysporum and recombinant Saccharomyces cerevisiae to efficiently cooperate in decomposition and ethanolic fermentation of wheat straw

Fusarium oxysporum F3 alone or in mixed culture with Saccharomyces cerevisiae F12 were used to ferment carbohydrates of wet exploded pre-treated wheat straw (PWS) directly to ethanol. Both microorganisms were first grown aerobically to produce cell mass and thereafter fermented PWS to ethanol under anaerobic conditions. During fermentation, soluble and insoluble carbohydrates were hydrolysed by the lignocellulolytic system of F. oxysporum. Mixed substrate fermentation using PWS and corn cobs (CC) in the ratio 1:2 was used to obtain an enzyme mixture with high cellulolytic and hemicellulolytic activities. Under these conditions, activities as high as 34300, 9100, 326, 24, 169, 27 and 254 U dm⁻³ of xylanase, endoglucanase, ??-glucosidase, arabinofuranosidase, avicelase, feruloyl esterase and acetyl esterase, respectively, were obtained. The replacement of the enzyme production phase of F. oxysporum by the addition of commercially available enzymes Celluclast^® 1.5 L FG and Novozym^® 188 in 3:1 ratio for the treatment of PWS, resulted in a 3-fold increase in the volumetric ethanol productivity without increasing the ethanol production significantly. By direct bioconversion of 110 kg m⁻³ dry matter of PWS, ethanol concentration (4.9 kg m⁻³) and yield (40 g kg⁻¹ of PWS) were similarly obtained by F. oxysporum and the mixed culture, while productivity rates as high as 34 g m⁻³ h⁻¹ and 108 g m⁻³ h⁻¹ were obtained by F. oxysporum and the mixed culture, respectively.     

Fuel ethanol production from sweet sorghum bagasse using microwave irradiation

Sweet sorghum is a hardy crop that can be grown on marginal land and can provide both food and energy in an integrated food and energy system. Lignocellulose rich sweet sorghum bagasse (solid left over after starch and juice extraction) can be converted to bioethanol using a variety of technologies. The largest barrier to commercial production of fuel ethanol from lignocellulosic material remains the high processing costs associated with enzymatic hydrolysis and the use of acids and bases in the pretreatment step. In this paper, sweet sorghum bagasse was pretreated and hydrolysed in a single step using microwave irradiation. A total sugar yield of 820 g kg⁻¹ was obtained in a 50 g kg⁻¹ sulphuric acid solution in water, with a power input of 43.2 kJ g⁻¹ of dry biomass (i.e. 20 min at 180 W power setting). An ethanol yield based on total sugar of 480 g kg⁻¹ was obtained after 24 h of fermentation using a mixed culture of organisms. These results show the potential for producing as much as 0.252 m³ tonne⁻¹ or 33 m³ ha⁻¹ ethanol using only the lignocellulose part of the stalks, which is high enough to make the process economically attractive.