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.

Effect of Different Microbial Strains on Biological Pretreatment of Sugarcane Bagasse for Enzymatic Hydrolysis

Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.

Ethanol Production from Sugarcane Bagasse by Means of Enzymes Produced by Solid State Fermentation Method

Nowadays there is a growing interest in biofuel production in most countries because of the increasing concerns about hydrocarbon fuel shortage and global climate changes, also for enhancing agricultural economy and producing local needs for transportation fuel. Ethanol can be produced from biomass by the hydrolysis and sugar fermentation processes. In this study ethanol was produced without using expensive commercial enzymes from sugarcane bagasse. Alkali pretreatment was used to prepare biomass before enzymatic hydrolysis. The comparison between NaOH, KOH and Ca(OH)2 shows NaOH is more effective on bagasse. The required enzymes for biomass hydrolysis were produced from sugarcane solid state fermentation via two fungi: Trichoderma longibrachiatum and Aspergillus niger. The results show that the produced enzyme solution via A. niger has functioned better than T. longibrachiatum. Ethanol was produced by simultaneous saccharification and fermentation (SSF) with crude enzyme solution from T. longibrachiatum and Saccharomyces cerevisiae yeast. To evaluate this procedure, SSF of pretreated bagasse was also done using Celluclast 1.5L by Novozymes. The yield of ethanol production by commercial enzyme and produced enzyme solution via T. longibrachiatum was 81% and 50% respectively.

Enhancing Efficiency for Reducing Sugar from Cassava Bagasse by Pretreatment

Cassava bagasse is one of major biomass wastes in Thailand from starch processing industry, which contains high starch content of about 60%. The object of this study was to investigate the optimal condition for hydrothermally pretreating cassava baggasses with or without acid addition. The pretreated samples were measured reducing sugar yield directly or after enzymatic hydrolysis (alpha-amylase). In enzymatic hydrolysis, the highest reducing sugar content was obtained under hydrothermal conditions for at 125oC for 30 min. The result shows that pretreating cassava baggasses increased the efficiency of enzymatic hydrolysis. For acid hydrolysis, pretreating cassava baggasses with sulfuric acid at 120oC for 60 min gave a maximum reducing sugar yield. In this study, sulfuric acid had a greater capacity for hydrolyzing cassava baggasses than phosphoric acid. In comparison, dilute acid hydrolysis to provide a higher yield of reducing sugar than the enzymatic hydrolysis combined hydrothermal pretreatment. However, enzymatic hydrolysis in a combination with hydrothermal pretreatment was an alternative to enhance efficiency reducing sugar production from cassava bagasse.

Effect of Temperature and Time on Dilute Acid Pretreatment of Corn Cobs

Lignocellulosic materials are new targeted source to produce second generation biofuels like biobutanol. However, this process is significantly resisted by the native structure of biomass. Therefore, pretreatment process is always essential to remove hemicelluloses and lignin prior to the enzymatic hydrolysis. The goals of pretreatment are removing hemicelluloses and lignin, increasing biomass porosity, and increasing the enzyme accessibility. The main goal of this research is to study the important variables such as pretreatment temperature and time, which can give the highest total sugar yield in pretreatment step by using dilute phosphoric acid. After pretreatment, the highest total sugar yield of 13.61 g/L was obtained under an optimal condition at 140°C for 10 min of pretreatment time by using 1.75% (w/w) H3PO4 and at 15:1 liquid to solid ratio. The total sugar yield of two-stage process (pretreatment+enzymatic hydrolysis) of 27.38 g/L was obtained.

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.