Search results for: Saccharification.

Authors: M. A. Kassim, R. Potumarthi, A. Tanksale, S. C. Srivatsa, S. Bhattacharya

Abstract:

Enzymatic saccharification of biomass for reducing sugar production is one of the crucial processes in biofuel production through biochemical conversion. In this study, enzymatic saccharification of dilute potassium hydroxide (KOH) pre-treated Tetraselmis suecica biomass was carried out by using cellulase enzyme obtained from Trichoderma longibrachiatum. Initially, the pre-treatment conditions were optimised by changing alkali reagent concentration, retention time for reaction, and temperature. The T. suecica biomass after pre-treatment was also characterized using Fourier Transform Infrared Spectra and Scanning Electron Microscope. These analyses revealed that the functional group such as acetyl and hydroxyl groups, structure and surface of T. suecica biomass were changed through pre-treatment, which is favourable for enzymatic saccharification process. Comparison of enzymatic saccharification of untreated and pre-treated microalgal biomass indicated that higher level of reducing sugar can be obtained from pre-treated T. suecica. Enzymatic saccharification of pre-treated T. suecica biomass was optimised by changing temperature, pH, and enzyme concentration to solid ratio ([E]/[S]). Highest conversion of carbohydrate into reducing sugar of 95% amounted to reducing sugar yield of 20 (wt%) from pre-treated T. suecica was obtained from saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1 after 72 h of incubation. Hydrolysate obtained from enzymatic saccharification of pretreated T. suecica biomass was further fermented into biobutanol using Clostridium saccharoperbutyliticum as biocatalyst. The results from this study demonstrate a positive prospect of application of dilute alkaline pre-treatment to enhance enzymatic saccharification and biobutanol production from microalgal biomass.

Keywords: Microalgal biomass, enzymatic saccharification, biobutanol, fermentation.

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Authors: Jhalique Jane R. Fojas, Ernesto J. Del Rosario

Abstract:

The dilute acid pretreatment and enzymatic saccharification of lignocellulosic substrate, cogon grass (Imperata cylindrical, L.) was optimized prior ethanol fermentation using simultaneous saccharification and fermentation (SSF) method. The optimum pretreatment conditions, temperature, sulfuric acid concentration, and reaction time were evaluated by determining the maximum sugar yield at constant enzyme loading. Cogon grass, at 10% w/v substrate loading, has optimum pretreatment conditions of 126°C, 0.6% v/v H2SO4, and 20min reaction time. These pretreatment conditions were used to optimize enzymatic saccharification using different enzyme combinations. The maximum saccharification yield of 36.68mg/mL (71.29% reducing sugar) was obtained using 25FPU/g-cellulose cellulase complex combined with 1.1% w/w of cellobiase, ß-glucosidase, and 0.225% w/w of hemicellulase complex, after 96 hours of saccharification. Using the optimum pretreatment and saccharification conditions, SSF of treated substrates was done at 37°C for 120 hours using industrial yeast strain HBY3, Saccharomyces cerevisiae. The ethanol yield for cogon grass at 4% w/w loading was 9.11g/L with 5.74mg/mL total residual sugar.

Keywords: Acid pretreatment, bioethanol, biomass, cogon grass, fermentation, lignocellylose, SSF.

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Authors: Rashid S. S., Alam M. Z.

Abstract:

A statistical optimization of the saccharification process of EFB was studied. The statistical analysis was done by applying faced centered central composite design (FCCCD) under response surface methodology (RSM). In this investigation, EFB dose, enzyme dose and saccharification period was examined, and the maximum 53.45% (w/w) yield of reducing sugar was found with 4% (w/v) of EFB, 10% (v/v) of enzyme after 120 hours of incubation. It can be calculated that the conversion rate of cellulose content of the substrate is more than 75% (w/w) which can be considered as a remarkable achievement. All the variables, linear, quadratic and interaction coefficient, were found to be highly significant, other than two coefficients, one quadratic and another interaction coefficient. The coefficient of determination (R2) is 0.9898 that confirms a satisfactory data and indicated that approximately 98.98% of the variability in the dependent variable, saccharification of EFB, could be explained by this model.

Keywords: Face centered central composite design (FCCCD), Liquid state bioconversion (LSB), Palm oil mill effluent, Trichoderma reesei RUT C-30.

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Authors: W. Wanmolee, W. Sornlake, N. Laosiripojana, V. Champreda

Abstract:

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.

Keywords: Biomass, Cellulase, Chaetomiun glubosum, Saccharification.

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Authors: K. Manikandan, T. Viruthagiri

Abstract:

Studies on Simultaneous Saccharification and Fermentation (SSF) of corn flour, a major agricultural product as the substrate using starch digesting glucoamylase enzyme derived from Aspergillus niger and non starch digesting and sugar fermenting Saccharomyces cerevisiae in a batch fermentation. Experiments based on Central Composite Design (CCD) were conducted to study the effect of substrate concentration, pH, temperature, enzyme concentration on Ethanol Concentration and the above parameters were optimized using Response Surface Methodology (RSM). The optimum values of substrate concentration, pH, temperature and enzyme concentration were found to be 160 g/l, 5.5, 30°C and 50 IU respectively. The effect of inoculums age on ethanol concentration was also investigated. The corn flour solution equivalent to 16% initial starch concentration gave the highest ethanol concentration of 63.04 g/l after 48 h of fermentation at optimum conditions of pH and temperature. Monod model and Logistic model were used for growth kinetics and Leudeking – Piret model was used for product formation kinetics.

Keywords: Simultaneous Saccharification and Fermentation(SSF), Corn Starch, Ethanol, Logisitic Model.

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Authors: Nasim Shaibani, Saba Ghazvini, Mohammad R. Andalibi, Soheila Yaghmaei

Abstract:

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.

Keywords: Alkali pretreatment, bioethanol, cellulase, simultaneous saccharification and fermentation, solid statefermentation, sugarcane bagasse

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Authors: E.Sasikumar, T.Viruthagiri

Abstract:

Simultaneous Saccharification and Fermentation (SSF) of sugarcane bagasse by cellulase and Pachysolen tannophilus MTCC *1077 were investigated in the present study. Important process variables for ethanol production form pretreated bagasse were optimized using Response Surface Methodology (RSM) based on central composite design (CCD) experiments. A 23 five level CCD experiments with central and axial points was used to develop a statistical model for the optimization of process variables such as incubation temperature (25–45°) X1, pH (5.0–7.0) X2 and fermentation time (24–120 h) X3. Data obtained from RSM on ethanol production were subjected to the analysis of variance (ANOVA) and analyzed using a second order polynomial equation and contour plots were used to study the interactions among three relevant variables of the fermentation process. The fermentation experiments were carried out using an online monitored modular fermenter 2L capacity. The processing parameters setup for reaching a maximum response for ethanol production was obtained when applying the optimum values for temperature (32°C), pH (5.6) and fermentation time (110 h). Maximum ethanol concentration (3.36 g/l) was obtained from 50 g/l pretreated sugarcane bagasse at the optimized process conditions in aerobic batch fermentation. Kinetic models such as Monod, Modified Logistic model, Modified Logistic incorporated Leudeking – Piret model and Modified Logistic incorporated Modified Leudeking – Piret model have been evaluated and the constants were predicted.

Keywords: Sugarcane bagasse, ethanol, optimization, Pachysolen tannophilus.

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Authors: Anita Singh, Namita Singh, Narsi R. Bishnoi

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To investigate the production of cellulases from Aspergillus heteromorphus, submerged fermentation was performed using wheat straw as substrate. Optimization of saccharification conditions like pH, temperature and time were studied. Highest reducing sugar was released on 5th day at 5 pH, 30° C temperature. When A. heteromorphous was grown on wheat straw in submerged fermentation after 5 days incubation at 30 ° C, 3.2 IU/ml and 83 IU/ml, filter paper activity and CMCase activity respectively.

Keywords: Aspergillus heteromorphus, Wheat Straw, Submerged Fermentation, Production of Cellulases

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Authors: Pilanee Vaithanomsat, Sinsupha Chuichulcherm, Waraporn Apiwatanapiwat

Abstract:

Sunflower stalks were analysed for chemical compositions: pentosan 15.84%, holocellulose 70.69%, alphacellulose 45.74%, glucose 27.10% and xylose 7.69% based on dry weight of 100-g raw material. The most optimum condition for steam explosion pretreatment was as follows. Sunflower stalks were cut into small pieces and soaked in 0.02 M H2SO4 for overnight. After that, they were steam exploded at 207 C and 21 kg/cm2 for 3 minutes to fractionate cellulose, hemicellulose and lignin. The resulting hydrolysate, containing hemicellulose, and cellulose pulp contained xylose sugar at 2.53% and 7.00%, respectively.The pulp was further subjected to enzymatic saccharification at 50 C, pH 4.8 citrate buffer) with pulp/buffer 6% (w/w)and Celluclast 1.5L/pulp 2.67% (w/w) to obtain single glucose with maximum yield 11.97%. After fixed-bed fermentation under optimum condition using conventional yeast mixtures to produce bioethanol, it indicated maximum ethanol yield of 0.028 g/100 g sunflower stalk.

Keywords: Enzymatic, steam explosion, sunflower stalk, ethanol production.

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Authors: A. Manivannan, R. T. Narendhirakannan

Abstract:

The objective of this research was to investigate biodegradation of water hyacinth (Eichhornia crassipes) to produce bioethanol using dilute-acid pretreatment (1% sulfuric acid) results in high hemicellulose decomposition and using yeast (Pachysolen tannophilus) as bioethanol producing strain. A maximum ethanol yield of 1.14g/L with coefficient, 0.24g g^-1; productivity, 0.015g l^-1h^-1 was comparable to predicted value 32.05g/L obtained by Central Composite Design (CCD). Maximum ethanol yield coefficient was comparable to those obtained through enzymatic saccharification and fermentation of acid hydrolysate using fully equipped fermentor. Although maximum ethanol concentration was low in lab scale, the improvement of lignocellulosic ethanol yield is necessary for large scale production.

Keywords: Acid hydrolysis, Biodegradation, Hemicellulose, Pachysolen tannophilus, Water hyacinth.

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Authors: Rotchanaphan Hengaroonprasan, Malinee Sriariyanun, Prapakorn Tantayotai, Supacharee Roddecha, Kraipat Cheenkachorn

Abstract:

Lignocellolusic material is a substance that is resistant to be degraded by microorganisms or hydrolysis enzymes. To be used as materials for biofuel production, it needs pretreatment process to improve efficiency of hydrolysis. In this work, chemical pretreatments on rice straw using three diluted organic acids, including acetic acid, citric acid, oxalic acid, were optimized. Using Response Surface Methodology (RSM), the effect of three pretreatment parameters, acid concentration, treatment time, and reaction temperature, on pretreatment efficiency were statistically evaluated. The results indicated that dilute oxalic acid pretreatment led to the highest enhancement of enzymatic saccharification by commercial cellulase and yielded sugar up to 10.67 mg/ml when using 5.04% oxalic acid at 137.11 oC for 30.01 min. Compared to other acid pretreatment by acetic acid, citric acid, and hydrochloric acid, the maximum sugar yields are 7.07, 6.30, and 8.53 mg/ml, respectively. Here, it was demonstrated that organic acids can be used for pretreatment of lignocellulosic materials to enhance of hydrolysis process, which could be integrated to other applications for various biorefinery processes. 

Keywords: Lignocellolusic biomass, pretreatment, organic acid response surface methodology, biorefinery.

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Authors: P. Martínez-Nieto, M. Vanegas-Hoyos, M. Zapata-Pineda, J. Robles-Camargo

Abstract:

The aquatic plants are a promising renewable energy resource. Lake Fúquene polluting macrophytes, water hyacinth (Eichhornia crassipes C. Mart.) and Brazilian elodea (Egeria densa Planch.), were saccharifiedby different treatments and fermented to ethanol by native yeasts. Among the tested chemical and biological methods for the saccharification, Pleurotus ostreatus at 10% (m/v) was chosen as the best pre-treatment in both macrophytes (P<0.01). Subsequently 49 yeasts were isolated from Lake Fúquene and nine strains were selected, which presented the highest precipitates characteristic of ethanol in the iodoform test. The fermentations from water hyacinth and Brazilian elodea hydrolysates using these yeasts produced ethanol at a rate between 0.38 to 0.80gL-1h-1 and 0.15 to 0.27gL-1h-1 respectively. The ethanol presence was confirmed by gas chromatography–mass spectrometry. The nine yeasts chosen were preliminarily identified as belonging to the genera Candida spp., Brettanomyces sp. and Hansenula spp.

Keywords: Bio-ethanol, Chemical hydrolysis, Invasive aquatic macrophytes, Native yeasts fermenting, P. ostreatus

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Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

Abstract:

The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever growing accepted option in the way to the development of biorefinery complexes. In the Mexican state of Sinaloa, a significant amount of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second generation bioethanol (2G), enzymes and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our work group. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU/mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: Biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol.

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