Surface carbohydrate analysis and bioethanol production of sugarcane bagasse pretreated with the white rot fungus, Ceriporiopsis subvermispora and microwave hydrothermolysis

Effects of pretreatments with a white rot fungus, Ceriporiopsis subvermispora, and microwave hydrothermolysis of bagasse on enzymatic saccharification and fermentation were evaluated. The best sugar yield, 44.9 g per 100 g of bagasse was obtained by fungal treatments followed by microwave hydrothermolysis at 180 °C for 20 min. Fluorescent-labeled carbohydrate-binding modules which recognize crystalline cellulose (CjCBM3-GFP), non-crystalline cellulose (CjCBM28-GFP) and xylan (CtCBM22-GFP) were applied to characterize the exposed polysaccharides. The microwave pretreatments with and without the fungal cultivation resulted in similar levels of cellulose exposure, but the combined treatment caused more defibration and thinning of the plant tissues. Simultaneous saccharification and fermentation of the pulp fractions obtained by microwave hydrothermolysis with and without fungal treatment, gave ethanol yields of 35.8% and 27.0%, respectively, based on the holocellulose content in the pulp. These results suggest that C. subvermispora pretreatment could be beneficial part of the process to produce ethanol from bagasse.     

Alkali-explosion pretreatment of straw and bagasse for enzymic hydrolysis

Sugarcane bagasse and wheat straw were subjected to alkali treatment at 200 degrees C for 5 min and at 3.45 MPa gas pressure (steam and nitrogen), followed by an explosive discharge through a defibrating nozzle, in an attempt to improve the rate and extent of digestibility. The treatment resulted in the solubilization of 40-45% of the components and in the production of a pulp that gave saccharification yields of 80 and 65% in 8 h for bagasse and wheat straw, respectively. By comparison, alkali steaming at 200 degrees C (1.72 MPa) for 5 min gave saccharification yields of only 58 and 52% in 48 h. The increase in temperature from 140 to 200 degrees C resulted in a gradual increase in in vitro organic matter digestibility (IVOMD) for both the substrates. Also, the extent of alkalinity during pretreatment appears to effect the reactivity of the final product towards enzymes. Pretreatment times ranging from 5 to 60 caused a progressive decline in the IVOMD of bagasse and wheat straw by the alkali explosion method and this was accompanied by a progressive decrease in pH values after explosion. In the alkali-steaming method, pretreatment time had no apparent effect with either substrate. An analysis of the alkali-exploded products showed that substantial amounts of hemicellulose and a small proportion of the lignin were solubilized. The percentage crystallinity of the cellulose did not alter in either substrate but there was a substantial reduction in the degree of polymerization. The superiority of the alkali-explosion pretreatment is attributed to the efficacy of fiber separation and disintegration; this increases the surface area and reduces the degree of polymerization.     

Radiation and fermentation treatment of cellulosic wastes

The effect of radiation pasteurization of sugar cane bagasse and rice straw and fermentation using various strains of fungi were studied for upgrading of cellulosic wastes. The initial contamination by fungi and aerobic bacteria both in bagasse and straw was high. The doses of 30 kGy for sterilization and 8 kGy for elimination of fungi were required. Irradiation effect showed that rice straw contained comparatively radioresistant microorganisms. It was observed that all the fungi (Hericium erinacium, Pleurotus djamor, Ganoderma lucidum, Auricularia auricula, Lentinus sajor-caju, Coriolus versicolor, Polyporus arcularius, Coprinus cinereus) grow extending over the entire substrates during one month after inoculation in irradiated bagasse and rice straw with 3% rice bran and 65% moisture content incubated at 30°C. Initially, sugar cane bagasse and rice straw substrates contained 39.4% and 25.9% of cellulose, 22.9% and 26.9% of hemicellulose, and 19.6% and 13.9% of lignin + cutin, respectively. Neutral detergent fibre (NDF) values decreased significantly in sugar cane bagasse fermented byG. lucidum, A. auricula andP. arcularius, and in rice straw fermented by all the 8 strains of fungi. Acid detergent fibre (ADF) values also decreased in bagasse and rice straw fermented by all the fungi.P. arcularius, H. erinacium, G. lucidum andC. cinereus were found to be the most effective strains for delignification of sugar cane bagasse.     

Preparation of levoglucosenone through sulfuric acid promoted pyrolysis of bagasse at low temperature

Fast pyrolysis of bagasse pretreated by sulfuric acid was conducted in a fixed bed reactor to prepare levoglucosenone (LGO), a very important anhydrosugar for organic synthesis. The liquid yield and LGO yield were studied at temperatures from 240 to 350 °C and sulfuric acid loadings from 0.92 to 7.10 wt.%. An optimal LGO yield of 7.58 wt.% was obtained at 270 °C with a sulfuric acid pretreatment concentration of 0.05 M (corresponding to 4.28 wt.% sulfuric acid loading). For comparison, microcrystalline cellulose pretreated by 0.05 M sulfuric acid solution was pyrolyzed at temperature from 270 °C to 320 °C, and bagasse loaded with 3-5 wt.% phosphoric acid was pyrolyzed at temperature from 270 °C to 350 °C. The highest yield of LGO from bagasse was 30% higher than that from microcrystalline cellulose, and treatment with sulfuric acid allowed a 21% higher yield than treatment with phosphoric acid.     

Production and characterization of xylanase from Bacillus thermoalkalophilus grown on agricultural wastes

Summary Bacillus thermoalkalophilus isolated from termite-infested mound soils of the semi-arid zones of India had the ability to produce good amounts of xylanase(s) from cheap agricultural wastes. Of the two hemicellulosic substrates tested, bagasse was found to be the better inducer for xylanase production. Alkali treatment of bagasse and rice husk had varied effects on enzyme production. The enzyme preparation had activity optima at 60° C and 70° C and a half-life of 60 min at 65° C. The enzyme was stable for 24 h over a pH range of 4.0–6.0, while maximum activity was observed at pH 6.0–7.0. Enzyme production and activity were inhibited by the end-product of xylan hydrolysis, xylose. Offprint requests to: Ajit Varma     

Effect of hot-water extraction on alkaline pulping of bagasse

The effect of hot-water extraction on alkaline pulping was investigated. The properties of black liquor and pulp strength of bagasse were analyzed. The extraction was conducted at 160 °C for 30 min where 13.2% of the mass was dissolved in the extraction liquor. Untreated bagasse and extracted bagasse were digested by soda and soda-AQ processes at 17% and 15.5% (with 0.1% AQ) alkali charge (NaOH). Cooking temperatures were 160 °C and 155 °C respectively. The pulp from extracted bagasse had a lower Kappa number and a higher viscosity compared to the pulp from the untreated bagasse. The black liquor from pulping extracted bagasse had a lower solid content, a lower viscosity and a lower silica content, but a higher heating value than that from pulping of untreated bagasse. Hot-water extraction resulted in a significant decrease in bleaching chemical consumption and the formation of chlorinated organics. Pulp strength properties such as the tensile index and the burst index were found to be lower, but the tear index, bulk, opacity and pulp freeness were found to be higher when hot-water extraction was applied.     

Optimization of microwave-assisted FeCl3 pretreatment conditions of rice straw and utilization of Trichoderma viride and Bacillus pumilus for production of reducing sugars

In this study, Box-Behnken design (BBD) and response surface methodology (RSM) were used to optimize microwave-assisted FeCl₃ pretreatment conditions of rice straw with respect to FeCl₃ concentration, microwave intensity, irradiation time and substrate concentration. When rice straw was pretreated at the optimal conditions of FeCl₃ concentration, 0.14 mol/L; microwave intensity, 160 °C; irradiation time, 19 min; substrate concentration, 109 g/L; and inoculated with Trichoderma viride and Bacillus pumilus, the production of reducing sugars was 6.62 g/L. This yield was 2.9 times higher than that obtained with untreated rice straw. The microorganisms degraded 37.8% of pretreated rice straw after 72 h. The structural characteristic analyses suggest that microwave-assisted FeCl₃ pretreatment damaged the silicified waxy surface of rice straw, disrupted almost all the ether linkages between lignin and carbohydrates, and removed lignin.     

Production of cellulase-free thermostable xylanases by an isolated strain of Aspergillus niger PPI, utilizing various lignocellulosic wastes

A strain of Aspergillus niger PPI having prolific xylanolytic potential was isolated and the optimum conditions for maximum xylanase production was studied, resulting in the following: 4% substrate concentration, 10% v/v inoculum size, 72 h of incubation and pH 3.5–4.5 at 28 °C. The production profile of xylanase was examined with various lignocellulosics and maximum yield was achieved with oat. The hemicellulose content of wastes was also determined and oatmeal was found to have maximum hemicellulose content followed by wheat straw, sugarcane bagasse, rice husk and gram residue respectively. The enzyme showed maximum activity at pH 4 and temperature 60 °C. However, maximum stability was achieved at pH 3.5 and temperature 55 °C. Cellulase activity was found altogether absent in the enzyme broth.     

Production of a cellulase-free xylanase from agricultural waste materials by a thermotolerant Streptomyces sp.

1444 microorganisms were isolated from soil samples from the northern Thai and screened at 55 °C by using basal medium supplemented with 1% carboxymethyl cellulose as a sole carbon source. One isolate, Streptomyces Ab106, had a high activity of a cellulase-free xylanase also without mannanase activity. The maximum cellulase-free xylanase activities of 3.5, 3.3, 3.1 and 2.7 IU were after growth of the organism with 1% (w/v) corn hull, corncob, bagasse and oat spelt xylan, respectively, at 55 °C for 6 days, respectively. The activity was more than 5 times higher than that at 35 °C.     

Injection of air into the headspace improves fermentation of phosphoric acid pretreated sugarcane bagasse by Escherichia coli MM170

Microaeration (injecting air into the headspace) improved the fermentation of hemicellulose hydrolysates obtained from the phosphoric acid pretreatment of sugarcane bagasse at 170 °C for 10 min. In addition, with 10% slurries of phosphoric acid pretreated bagasse (180 °C, 10 min), air injection into the headspace promoted xylose utilization and increased ethanol yields from 0.16 to 0.20 g ethanol/g bagasse dry weight using a liquefaction plus simultaneous saccharification and co-fermentation process (L+SScF). This process was scaled up to 80 L using slurries of acid pretreated bagasse (96 h incubation; 0.6 L of air/min into the headspace) with ethanol yields of 312-347 L (82-92 gal) per tonne (dry matter), corresponding to 0.25 and 0.27 g/g bagasse (dry weight). Injection of small amounts of air into the headspace may provide a convenient alternative to subsurface sparging that avoids problems of foaming, sparger hygiene, flotation of particulates, and phase separation.     

Improved enzymic digestibility of sugar cane bagasse following high temperature autohydrolysis

Summary Sugar cane bagasse was subjected to a two stage autohydrolysis treatment. In the first stage bagasse was heated in the presence of varying amounts of water at temperatures in the range 160–180°C to extract the hemicellulose fraction. The water insoluble residue was then heated in the presence of varying amounts of water at temperatures in the range 203 to 241°C. The effect of autohydrolysis on the digestibility of bagasse was assessed by hydrolysing the material with Trichoderma reesei cellulase enzymes (0.65 FPU/ml).     

Optimization of H2SO4-catalyzed hydrothermal pretreatment of rapeseed straw for bioconversion to ethanol: Focusing on pretreatment at high solids content

A central composite design of response surface method was used to optimize H₂SO₄-catalyzed hydrothermal pretreatment of rapeseed straw, in respect to acid concentration (0.5–2%), treatment time (5–20 min) and solid content (10–20%) at 180 °C. Enzymatic hydrolysis and fermentation were also measured to evaluate the optimal pretreatment conditions for maximizing ethanol production. The results showed that acid concentration and treatment time were more significant than solid content for optimization of xylose release and cellulose recovery. Pretreatment with 1% sulfuric acid and 20% solid content for 10 min at 180 °C was found to be the most optimal condition for pretreatment of rapeseed straw for ethanol production. After pretreatment at the optimal condition and enzymatic hydrolysis, 75.12% total xylan and 63.17% total glucan were converted to xylose and glucose, respectively. Finally, 66.79% of theoretical ethanol yielded after fermentation.     

Explosive pretreatment of lignocellulosic residues with high-pressure carbon dioxide for the production of fermentation substrates

A new pretreatment technique has been developed in which the lignocellulosic material is subjected to the action of steam and high-pressure carbon dioxide before being explosively discharged through a defibrating nozzle of novel design. Operating at 200 degrees C and gas pressures in the range of 3.45-13.8 MPa, exploded products with maximum in vitro cellulase digestibilities of 81, 78, and 75% were obtained from wheat straw, bagasse, and Eucalyptus regnans woodchips, respectively. The treatment times required to obtain substrates of maximum digestibility were 5 min for wheat straw and bagasse and 15 min for E. regnans. Analysis of the exploded products indicated that the pretreatment had substantially removed and solubilized the hemicellulose fraction of the feed materials, giving an autohydrolysis liquor rich in xylose and a fibrous residue primarily composed of alpha-cellulose and lignin. The fibrous residue was readily amenable to cellulase hydrolysis, with saccharification being completed within 48 h. The theoretical energy demand for gas compression in the pretreatment process has been calculated at 11 k Wh/ton raw material when the digester is operated at 3.45 MPa and a packing density of 250 kg raw material/m(3) digester volume.     

Effects of temperature and holding time on production of renewable fuels from pyrolysis of Chlorella protothecoides

To investigate the influence of temperature andholding time on the pyrolyzate yields of Chlorella protothecoides, the microalgal cells weresubjected to pyrolysis at 200, 300, 400, 500 and 600 ^°Cfor 5, 20, 60 and 120 min, separately. High oil yields above 40% dry weight cells wereobtained both at relatively low temperature (300 ^°C)with relatively long holding times (20–120min) and relatively high temperatures (400–500 ^°C)with relatively short holding times (5–20min). The maximum oil yield of 52.0% was achieved at500 ^°C for 5 min. The gas yield was generallyincreased with the increasing temperature and holdingtime. It could reach 63.3–76.0% at 600 ^°C.High pyrolytic rates of 72–87% were obtained at allexperiments except at 200 ^°C for 5–20 min or300 ^°C for 5 min. Thermogravimetric analysisindicated that the main thermal degradation of thismicroalga occurred at 200–520 ^°C. The resultsimply that C. protothecoides is a good candidatefor the production of renewable fuels by pyrolysis.     

Evaluation of different alkali treatments of bagasse pith for cultivation of Cellulomonas sp.

Biomass production of Cellulomonas was optimal with 1% (w/v) bagasse pith pre-treated with either 0.2 M NaOH for 1 h at 80°C or 0.4 M NaOH for 40 h at 28 to 30°C. Growth was similar to that obtained with a more severe treatment used as control and compared well with other reports for cellulolytic bacteria cultivated on pre-treated bagasse pith.     

Pyrolysis of grape bagasse: effect of pyrolysis conditions on the product yields and characterization of the liquid product

In this study, pyrolysis of grape bagasse was investigated with the aim to study the product distribution and their chemical compositions and to identify optimum process conditions for maximizing the bio-oil yield. Particular investigated process variables were temperature (350-600 °C), heating rate (10-50 °C/min) and nitrogen gas flow rate (50-200 cm³/min). The maximum oil yield of 27.60% was obtained at the final pyrolysis temperature of 550 °C, sweeping gas flow rate of 100 cm³/min and heating rate of 50 °C/min in a fixed-bed reactor. The elemental analysis and heating value of the bio-oils were determined, and then the chemical composition of the bio-oil was investigated using chromatographic and spectroscopic techniques such as column chromatography, ¹H NMR and FTIR. The fuel properties of the bio-oil such as flash point, viscosity and density were also determined. The bio-oils obtained from grape bagasse were presented as an environmentally friendly feedstock candidate for bio-fuels.     

Optimisation of a microwave pretreatment of wheat straw for methane production

This study aims at the optimisation of a microwave pretreatment for wheat straw solubilisation and anaerobic biodegradability. The maximum yield of methane production was obtained at 150 °C with an improvement of 28% compared to an untreated sample. In addition, at this temperature, the time to reach 80% of the methane volume obtained from untreated straw was about 35%. The study of ramp time and holding time at targeted temperature showed that they had no improvement effect. Thus, the best conditions are the highest heating rate for a final temperature 150 °C without any holding time. The reading of energy consumed by pretreatment and energy overproduced by pretreated samples showed that increasing tVS amount and heating rate led to a saving of energy consumption. Nevertheless, to obtain a positive energy balance, a microwave device should consume less than 2.65 kJ/g_tVS.     

Sugar production from agricultural woody wastes by saccharification with Trichoderma viride cellulase.

The saccharification of agricultural woody wastes was studied using a commercial enzyme preparation, Cellulase onozuka, derived from Trichoderma viride or the solid culture extracts of the fungus. With the intention of producing sugar at low cost, a simple procedure of enzymatic saccharification of rice straw, bagasse, and sawdust was studied. Delignifying methods of these wastes were investigated using dilute sodium hydroxide solution and dilute peracetic acid. Rice straw and bagasse were effectively delignified by boiling in a 1% sodium hydroxide solution for 3 hr or by autoclaving at 120 degrees C in a 1% sodium hydroxide solution for 1 hr. The sawdust from a broad leaved tree (Machilus thunbergii) was delignified by autoclaving at 120 degrees C in a 1% sodium hydroxide solution for 1 hr and by subsequent boiling in diluted 1/5 peracetic acid for 1 hr. This type of sawdust was also delignified by boiling in 1/5 peracetic acid for 1 hr and by subsequent autoclaving at 120 degrees C in a 1% sodium hydroxide solution for 1 hr. The sawdust from a coniferous tree (Cryptomeria japonica) was delignified by boiling in 1/5 peracetic acid for 1 hr and by subsequent autoclaving at 120 degrees C in a 1% sodium hydroxide solution for 1 hr; however, the successive treatment by autoclaving with alkali solution and subsequent boiling with diluted peracetic acid failed to bring about the desired effect. The saccharification of delignified rice straw, bagasse, and sawdust was examined using Cellulase onozuka, wheat bran or rice straw solid culture at various substrate concentrations, resulting in the formation of 5 to 10% sugar solutions after incubation at pH 5.0, 45 degrees C for 48 hr. The optimum substrate concentration existed at around 10%. Reuse of cellulase solution and resaccharification of residual sawdust were considered to be inadequate.     

Cellulolytic enzymes of a thermotolerantAspergillus terreus strain and their action on cellulosic substrates

A thermotolerant fungal strainAspergillus terreus produced high activities of cellulolytic enzymes when grown in shake flasks for 8 days at 40°C or 14 days at 28°C in medium containing 2.5% (w/v) cellulose powder and 1% (w/v) wheat bran. There was little difference between the final activities of endo-(1,4)--glucanase (ca. 14.4 U/ml); filter paper activity (ca. 1.3 U/ml) and -glucosidase (ca. 10 U/ml). Endoglucanase had maximum activity at 60°C and pH 3.8; the other two enzymes were optimal at 60°C and pH 4.8. The maximum hydrolysis of different cellulosic substrates (about 50%) was obtained within 48 h when 1.1 U/ml of filter paper cellulase activity were employed to saccharify 100 mg alkali-treated cotton, filter paper, bagasse, and rice straw at 50°C and pH 4.8. The major end-product, glucose, was produced from all substrates, with traces of cellobiose and other larger oligosaccharides being present in rice straw hydrolysates.     

Liquid hot water and alkaline pretreatment of soybean straw for improving cellulose digestibility

Soybean straw was pretreated with either liquid hot water (LHW) (170-210 °C for 3-10 min) or alkaline soaking (4-40 g NaOH/100 g dry straw) at room temperature to evaluate the effects on cellulose digestibility. Nearly 100% cellulose was recovered in pretreated solids for both pretreatment methods. For LHW pretreatment, xylan dissolution from the raw material increased with pretreatment temperature and time. Cellulose digestibility was correlated with xylan dissolution. A maximal glucose yield of 70.76%, corresponding to 80% xylan removal, was obtained with soybean straw pretreated at 210 °C for 10 min. NaOH soaking at ambient conditions removed xylan up to 46.37% and the subsequent glucose yield of pretreated solids reached up to 64.55%. Our results indicated LHW pretreatment was more effective than NaOH soaking for improving cellulose digestibility of soybean straw.