Characterisation of Fractions Extracted from Sorghum Byproducts

Sorghum byproducts, namely bran, stalk, and panicle are examples of lignocellulosic biomass. These raw materials contain large amounts of polysaccharides, in particular hemicelluloses, celluloses, and lignins, which if efficiently extracted, can be utilised for the development of a range of added value products with potential applications in agriculture and food packaging sectors. The aim of this study was to characterise fractions extracted from sorghum bran and stalk with regards to their physicochemical properties that could determine their applicability as food-packaging materials. A sequential alkaline extraction was applied for the isolation of cellulosic, hemicellulosic and lignin fractions from sorghum stalk and bran. Lignin content, phenolic content and antioxidant capacity were also investigated in the case of the lignin fraction. Thermal analysis using differential scanning calorimetry (DSC) and X-Ray Diffraction (XRD) revealed that the glass transition temperature (Tg) of cellulose fraction of the stalk was ~78.33 oC at amorphous state (~65%) and water content of ~5%. In terms of hemicellulose, the Tg value of stalk was slightly lower compared to bran at amorphous state (~54%) and had less water content (~2%). It is evident that hemicelluloses generally showed a lower thermal stability compared to cellulose, probably due to their lack of crystallinity. Additionally, bran had higher arabinose-to-xylose ratio (0.82) than the stalk, a fact that indicated its low crystallinity. Furthermore, lignin fraction had Tg value of ~93 oC at amorphous state (~11%). Stalk-derived lignin fraction contained more phenolic compounds (mainly consisting of p-coumaric and ferulic acid) and had higher lignin content and antioxidant capacity compared to bran-derived lignin fraction.

Production and Purification of Monosaccharides by Hydrolysis of Sugar Cane Bagasse in an Ionic Liquid Medium

The conversion of lignocellulosic waste materials, such as sugar cane bagasse, to biofuels such as ethanol has attracted significant interest as a potential element for transforming transport fuel supplies to totally renewable sources. However, the refractory nature of the cellulosic structure of lignocellulosic materials has impeded progress on developing an economic process, whereby the cellulose component may be effectively broken down to glucose monosaccharides and then purified to allow downstream fermentation. Ionic liquid (IL) treatment of lignocellulosic biomass has been shown to disrupt the crystalline structure of cellulose thus potentially enabling the cellulose to be more readily hydrolysed to monosaccharides. Furthermore, conventional hydrolysis of lignocellulosic materials yields byproducts that are inhibitors for efficient fermentation of the monosaccharides. However, selective extraction of monosaccharides from an aqueous/IL phase into an organic phase utilizing a combination of boronic acids and quaternary amines has shown promise as a purification process. Hydrolysis of sugar cane bagasse immersed in an aqueous solution with IL (1-ethyl-3-methylimidazolium acetate) was conducted at different pH and temperature below 100 ºC. It was found that the use of a high concentration of hydrochloric acid to acidify the solution inhibited the hydrolysis of bagasse. At high pH (i.e. basic conditions), using sodium hydroxide, catalyst yields were reduced for total reducing sugars (TRS) due to the rapid degradation of the sugars formed. For purification trials, a supported liquid membrane (SLM) apparatus was constructed, whereby a synthetic solution containing xylose and glucose in an aqueous IL phase was transported across a membrane impregnated with phenyl boronic acid/Aliquat 336 to an aqueous phase. The transport rate of xylose was generally higher than that of glucose indicating that a SLM scheme may not only be useful for purifying sugars from undesirable toxic compounds, but also for fractionating sugars to improve fermentation efficiency.

Screening of Factors Affecting the Enzymatic Hydrolysis of Empty Fruit Bunches in Aqueous Ionic Liquid and Locally Produced Cellulase System

The enzymatic hydrolysis of lignocellulosic biomass is one of the obstacles in the process of sugar production, due to the presence of lignin that protects the cellulose molecules against cellulases. Although the pretreatment of lignocellulose in ionic liquid (IL) system has been receiving a lot of interest; however, it requires IL removal with an anti-solvent in order to proceed with the enzymatic hydrolysis. At this point, introducing a compatible cellulase enzyme seems more efficient in this process. A cellulase enzyme that was produced by Trichoderma reesei on palm kernel cake (PKC) exhibited a promising stability in several ILs. The enzyme called PKC-Cel was tested for its optimum pH and temperature as well as its molecular weight. One among evaluated ILs, 1,3-diethylimidazolium dimethyl phosphate [DEMIM] DMP was applied in this study. Evaluation of six factors was executed in Stat-Ease Design Expert V.9, definitive screening design, which are IL/ buffer ratio, temperature, hydrolysis retention time, biomass loading, cellulase loading and empty fruit bunches (EFB) particle size. According to the obtained data, IL-enzyme system shows the highest sugar concentration at 70 °C, 27 hours, 10% IL-buffer, 35% biomass loading, 60 Units/g cellulase and 200 μm particle size. As concluded from the obtained data, not only the PKC-Cel was stable in the presence of the IL, also it was actually stable at a higher temperature than its optimum one. The reducing sugar obtained was 53.468±4.58 g/L which was equivalent to 0.3055 g reducing sugar/g EFB. This approach opens an insight for more studies in order to understand the actual effect of ILs on cellulases and their interactions in the aqueous system. It could also benefit in an efficient production of bioethanol from lignocellulosic biomass.

Analysis of a Lignocellulose Degrading Microbial Consortium to Enhance the Anaerobic Digestion of Rice Straws

Rice straw is lignocellulosic biomass which can be utilized as substrate for the biogas production. However, due to the property and composition of rice straw, it is difficult to be degraded by hydrolysis enzymes. One of the pretreatment methods that modify such properties of lignocellulosic biomass is the application of lignocellulose-degrading microbial consortia. The aim of this study is to investigate the effect of microbial consortia to enhance biogas production. To select the high efficient consortium, cellulase enzymes were extracted and their activities were analyzed. The results suggested that microbial consortium culture obtained from cattle manure is the best candidate compared to decomposed wood and horse manure. A microbial consortium isolated from cattle manure was then mixed with anaerobic sludge and used as inoculum for biogas production. The optimal conditions for biogas production were investigated using response surface methodology (RSM). The tested parameters were the ratio of amount of microbial consortium isolated and amount of anaerobic sludge (MI:AS), substrate to inoculum ratio (S:I) and temperature. Here, the value of the regression coefficient R2 = 0.7661 could be explained by the model which is high to advocate the significance of the model. The highest cumulative biogas yield was 104.6 ml/g-rice straw at optimum ratio of MI:AS, ratio of S:I, and temperature of 2.5:1, 15:1 and 44°C respectively.

Application of Statistical Approach for Optimizing CMCase Production by Bacillus tequilensis S28 Strain via Submerged Fermentation Using Wheat Bran as Carbon Source

Biofuels production has come forth as a future technology to combat the problem of depleting fossil fuels. Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient method and catching the eye of scientific community. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus main objective of the present study was to optimize composition of medium components for enhancing cellulase production by newly isolated strain of Bacillus tequilensis. Nineteen factors were taken into account using statistical Plackett-Burman Design. The significant variables influencing the cellulose production were further employed in statistical Response Surface Methodology using Central Composite Design for maximizing cellulase production. The optimum medium composition for cellulase production was: peptone (4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L), Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride (0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at 37oC. Analysis of variance (ANOVA) revealed high coefficient of determination (R2) of 0.99. Maximum cellulase productivity of 11.5 IU/ml was observed against the model predicted value of 13 IU/ml. This was found to be optimally active at 60oC and pH 5.5.

Conditions of the Anaerobic Digestion of Biomass

Biological conversion of biomass to methane has received increasing attention in recent years. Grasses have been explored for their potential anaerobic digestion to methane. In this review, extensive literature data have been tabulated and classified. The influences of several parameters on the potential of these feedstocks to produce methane are presented. Lignocellulosic biomass represents a mostly unused source for biogas and ethanol production. Many factors, including lignin content, crystallinity of cellulose, and particle size, limit the digestibility of the hemicellulose and cellulose present in the lignocellulosic biomass. Pretreatments have used to improve the digestibility of the lignocellulosic biomass. Each pretreatment has its own effects on cellulose, hemicellulose and lignin, the three main components of lignocellulosic biomass. Solidstate anaerobic digestion (SS-AD) generally occurs at solid concentrations higher than 15%. In contrast, liquid anaerobic digestion (AD) handles feedstocks with solid concentrations between 0.5% and 15%. Animal manure, sewage sludge, and food waste are generally treated by liquid AD, while organic fractions of municipal solid waste (OFMSW) and lignocellulosic biomass such as crop residues and energy crops can be processed through SS-AD. An increase in operating temperature can improve both the biogas yield and the production efficiency, other practices such as using AD digestate or leachate as an inoculant or decreasing the solid content may increase biogas yield but have negative impact on production efficiency. Focus is placed on substrate pretreatment in anaerobic digestion (AD) as a means of increasing biogas yields using today’s diversified substrate sources.

Development of Efficient Fungal Biomass-Degrading Enzyme Mixtures for Saccharification of Local Lignocellulosic Feedstock

Conversion of lignocellulosic biomass is the basis process for production of fuels, chemicals and materials in the sustainable biorefinery industry. Saccharification of lignocellulosic biomass is an essential step which produces sugars for further conversion to target value-added products e.g. bio-ethanol, bio-plastic, g-valerolactone (GVL), 5-hydroxymethylfuroic acid (HMF), levulinic acid, etc. The goal of this work was to develop an efficient enzyme for conversion of biomass to reducing sugar based on crude fungal enzyme from Chaetomium globosum BCC5776 produced by submerged fermentation and evaluate its activity comparing to a commercial Acremonium cellulase. Five local biomasses in Thailand: rice straw, sugarcane bagasse, corncobs, corn stovers, and palm empty fruit bunches were pretreated and hydrolyzed with varying enzyme loadings. Saccharification of the biomass led to different reducing sugar levels from 115 mg/g to 720 mg/g from different types of biomass using cellulase dosage of 9 FPU/g. The reducing sugar will be further employed as sugar feedstock for production of ethanol or commodity chemicals. This work demonstrated the use of promising enzyme candidate for conversion of local lignocellulosic biomass in biorefinery industry.

Thermal and Morphological Evaluation of Chemically Pretreated Sugarcane Bagasse

Enzymatic hydrolysis is one of the major steps involved in the conversion from sugarcane bagasse to yield ethanol. This process offers potential for yields and selectivity higher, lower energy costs and milder operating conditions than chemical processes. However, the presence of some factors such as lignin content, crystallinity degree of the cellulose, and particle sizes, limits the digestibility of the cellulose present in the lignocellulosic biomasses. Pretreatment aims to improve the access of the enzyme to the substrate. In this study sugarcane bagasse was submitted chemical pretreatment that consisted of two consecutive steps, the first with dilute sulfuric acid (1 % (v/v) H2SO4), and the second with alkaline solutions with different concentrations of NaOH (1, 2, 3 and 4 % (w/v)). Thermal Analysis (TG/ DTG and DTA) was used to evaluate hemicellulose, cellulose and lignin contents in the samples. Scanning Electron Microscopy (SEM) was used to evaluate the morphological structures of the in natura and chemically treated samples. Results showed that pretreatments were effective in chemical degradation of lignocellulosic materials of the samples, and also was possible to observe the morphological changes occurring in the biomasses after pretreatments.

Bioprocessing of Proximally Analyzed Wheat Straw for Enhanced Cellulase Production through Process Optimization with Trichodermaviride under SSF

The purpose of the present work was to study the production and process parameters optimization for the synthesis of cellulase from Trichoderma viride in solid state fermentation (SSF) using an agricultural wheat straw as substrates; as fungal conversion of lignocellulosic biomass for cellulase production is one among the major increasing demand for various biotechnological applications. An optimization of process parameters is a necessary step to get higher yield of product. Several kinetic parameters like pretreatment, extraction solvent, substrate concentration, initial moisture content, pH, incubation temperature and inoculum size were optimized for enhanced production of third most demanded industrially important cellulase. The maximum cellulase enzyme activity 398.10±2.43 μM/mL/min was achieved when proximally analyzed lignocellulosic substrate wheat straw inocubated at 2% HCl as pretreatment tool along with distilled water as extraction solvent, 3% substrate concentration 40% moisture content with optimum pH 5.5 at 45°C incubation temperature and 10% inoculum size.

Optimization of Two-Stage Pretreatment Combined with Microwave Radiation Using Response Surface Methodology

Pretreatment is an essential step in the conversion of lignocellulosic biomass to fermentable sugar that used for biobutanol production. Among pretreatment processes, microwave is considered to improve pretreatment efficiency due to its high heating efficiency, easy operation, and easily to combine with chemical reaction. The main objectives of this work are to investigate the feasibility of microwave pretreatment to enhance enzymatic hydrolysis of corncobs and to determine the optimal conditions using response surface methodology. Corncobs were pretreated via two-stage pretreatment in dilute sodium hydroxide (2 %) followed by dilute sulfuric acid 1 %. Pretreated corncobs were subjected to enzymatic hydrolysis to produce reducing sugar. Statistical experimental design was used to optimize pretreatment parameters including temperature, residence time and solid-to-liquid ratio to achieve the highest amount of glucose. The results revealed that solid-to-liquid ratio and temperature had a significant effect on the amount of glucose.

Thermogravimetry Study on Pyrolysis of Various Lignocellulosic Biomass for Potential Hydrogen Production

This paper aims to study decomposition behavior in pyrolytic environment of four lignocellulosic biomass (oil palm shell, oil palm frond, rice husk and paddy straw), and two commercial components of biomass (pure cellulose and lignin), performed in a thermogravimetry analyzer (TGA). The unit which consists of a microbalance and a furnace flowed with 100 cc (STP) min-1 Nitrogen, N2 as inert. Heating rate was set at 20⁰C min-1 and temperature started from 50 to 900⁰C. Hydrogen gas production during the pyrolysis was observed using Agilent Gas Chromatography Analyzer 7890A. Oil palm shell, oil palm frond, paddy straw and rice husk were found to be reactive enough in a pyrolytic environment of up to 900°C since pyrolysis of these biomass starts at temperature as low as 200°C and maximum value of weight loss is achieved at about 500°C. Since there was not much different in the cellulose, hemicelluloses and lignin fractions between oil palm shell, oil palm frond, paddy straw and rice husk, the T-50 and R-50 values obtained are almost similar. H2 productions started rapidly at this temperature as well due to the decompositions of biomass inside the TGA. Biomass with more lignin content such as oil palm shell was found to have longer duration of H2 production compared to materials of high cellulose and hemicelluloses contents.

Statistical Optimization of Process Variables for Direct Fermentation of 226 White Rose Tapioca Stem to Ethanol by Fusarium oxysporum

Direct fermentation of 226 white rose tapioca stem to ethanol by Fusarium oxysporum was studied in a batch reactor. Fermentation of ethanol can be achieved by sequential pretreatment using dilute acid and dilute alkali solutions using 100 mesh tapioca stem particles. The quantitative effects of substrate concentration, pH and temperature on ethanol concentration were optimized using a full factorial central composite design experiment. The optimum process conditions were then obtained using response surface methodology. The quadratic model indicated that substrate concentration of 33g/l, pH 5.52 and a temperature of 30.13oC were found to be optimum for maximum ethanol concentration of 8.64g/l. The predicted optimum process conditions obtained using response surface methodology was verified through confirmatory experiments. Leudeking-piret model was used to study the product formation kinetics for the production of ethanol and the model parameters were evaluated using experimental data.

Ionic Liquid Pretreatment and Enzymatic Hydrolysis of Wood Biomass

Pretreatment of lignocellulosic biomass materials from poplar, acacia, oak, and fir with different ionic liquids (ILs) containing 1-alkyl-3-methyl-imidazolium cations and various anions has been carried out. The dissolved cellulose from biomass was precipitated by adding anti-solvents into the solution and vigorous stirring. Commercial cellulases Celluclast 1.5L and Accelerase 1000 have been used for hydrolysis of untreated and pretreated lignocellulosic biomass. Among the tested ILs, [Emim]COOCH3 showed the best efficiency, resulting in highest amount of liberated reducing sugars. Pretreatment of lignocellulosic biomass using glycerol-ionic liquids combined pretreatment and dilute acid-ionic liquids combined pretreatment were evaluated and compared with glycerol pretreatment, ionic liquids pretreatment and dilute acid pretreatment.

Characterization of Corn Cobs from Microwave and Potassium Hydroxide Pretreatment

The complexity of lignocellulosic biomass requires a pretreatment step to improve the yield of fermentable sugars. The efficient pretreatment of corn cobs using microwave and potassium hydroxide and enzymatic hydrolysis was investigated. The objective of this work was to characterize the optimal condition of pretreatment of corn cobs using microwave and potassium hydroxide enhance enzymatic hydrolysis. Corn cobs were submerged in different potassium hydroxide concentration at varies temperature and resident time. The pretreated corn cobs were hydrolyzed to produce the reducing sugar for analysis. The morphology and microstructure of samples were investigated by Thermal gravimetric analysis (TGA, scanning electron microscope (SEM), X-ray diffraction (XRD). The results showed that lignin and hemicellulose were removed by microwave/potassium hydroxide pretreatment. The crystallinity of the pretreated corn cobs was higher than the untreated. This method was compared with autoclave and conventional heating method. The results indicated that microwave-alkali treatment was an efficient way to improve the enzymatic hydrolysis rate by increasing its accessibility hydrolysis enzymes.

A Comparison of Dilute Sulfuric and Phosphoric Acid Pretreatments in Biofuel Production from Corncobs

Biofuels, like biobutanol, have been recognized for being renewable and sustainable fuels which can be produced from lignocellulosic biomass. To convert lignocellulosic biomass to biofuel, pretreatment process is an important step to remove hemicelluloses and lignin to improve enzymatic hydrolysis. Dilute acid pretreatment has been successful developed for pretreatment of corncobs and the optimum conditions of dilute sulfuric and phosphoric acid pretreatment were obtained at 120 °C for 5 min with 15:1 liquid to solid ratio and 140 °C for 10 min with 10:1 liquid to solid ratio, respectively. The result shows that both of acid pretreatments gave the content of total sugar approximately 34–35 g/l. In case of inhibitor content (furfural), phosphoric acid pretreatment gives higher than sulfuric acid pretreatment. Characterizations of corncobs after pretreatment indicate that both of acid pretreatments can improve enzymatic accessibility and the better results present in corncobs pretreated with sulfuric acid in term of surface area, crystallinity, and composition analysis.

Pyrolysis Characteristics and Kinetics of Macroalgae Biomass Using Thermogravimetric Analyzer

The pyrolysis characteristics and kinetics of seven marine biomass, which are fixed Enteromorpha clathrata, floating Enteromorpha clathrata, Ulva lactuca L., Zosterae Marinae L., Thallus Laminariae, Asparagus schoberioides kunth and Undaria pinnatifida (Harv.), were studied with thermogravimetric analysis method. Simultaneously, cornstalk, which is a grass biomass, and sawdust, which is a lignocellulosic biomass, were references. The basic pyrolysis characteristics were studied by using TG- DTG-DTA curves. The results showed that there were three stages (dehydration, dramatic weight loss and slow weight loss) during the whole pyrolysis process of samples. The Tmax of marine biomass was significantly lower than two kinds of terrestrial biomass. Zosterae Marinae L. had a relatively high stability of pyrolysis, but floating Enteromorpha clathrata had lowest stability of pyrolysis and a good combustion characteristics. The corresponding activation energy E and frequency factor A were obtained by Coats-Redfern method. It was found that the pyrolysis reaction mechanism functions of three kinds of biomass are different.