Commenced in January 2007
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Edition: International
Paper Count: 49

Search results for: Enzymatic hydrolysis

49 Ionic Liquid Pretreatment and Enzymatic Hydrolysis of Wood Biomass

Authors: M. Ungurean, F. Fitigau, C. Paul, A. Ursoiu, F. Peter

Abstract:

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.

Keywords: Cellulase, enzymatic hydrolysis, lignocellulosicbiomass, pretreatment.

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48 Optimization of Enzymatic Hydrolysis of Manihot Esculenta Root Starch by Immobilizeda-Amylase Using Response Surface Methodology

Authors: G. Baskar, C. Muthukumaran, S. Renganathan

Abstract:

Enzymatic hydrolysis of starch from natural sources finds potential application in commercial production of alcoholic beverage and bioethanol. In this study the effect of starch concentration, temperature, time and enzyme concentration were studied and optimized for hydrolysis of cassava (Manihot esculenta) starch powder (of mesh 80/120) into glucose syrup by immobilized (using Polyacrylamide gel) a-amylase using central composite design. The experimental result on enzymatic hydrolysis of cassava starch was subjected to multiple linear regression analysis using MINITAB 14 software. Positive linear effect of starch concentration, enzyme concentration and time was observed on hydrolysis of cassava starch by a-amylase. The statistical significance of the model was validated by F-test for analysis of variance (p < 0.01). The optimum value of starch concentration temperature, time and enzyme concentration were found to be 4.5% (w/v), 45oC, 150 min, and 1% (w/v) enzyme. The maximum glucose yield at optimum condition was 5.17 mg/mL.

Keywords: Enzymatic hydrolysis, Alcoholic beverage, Centralcomposite design, Polynomial model, glucose yield.

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47 Study on Microbial Pretreatment for Enhancing Enzymatic Hydrolysis of Corncob

Authors: Kessara Seneesrisakul, Erdogan Gulari, Sumaeth Chavadej

Abstract:

The complex structure of lignocellulose leads to great difficulties in converting it to fermentable sugars for the ethanol production. The major hydrolysis impediments are the crystallinity of cellulose and the lignin content. To improve the efficiency of enzymatic hydrolysis, microbial pretreatment of corncob was investigated using two bacterial strains of Bacillus subtilis A 002 and Cellulomonas sp. TISTR 784 (expected to break open the crystalline part of cellulose) and lignin-degrading fungus, Phanerochaete sordida SK7 (expected to remove lignin from lignocellulose). The microbial pretreatment was carried out with each strain under its optimum conditions. The pretreated corncob samples were further hydrolyzed to produce reducing glucose with low amounts of commercial cellulase (25 U·g-1 corncob) from Aspergillus niger. The corncob samples were determined for composition change by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). According to the results, the microbial pretreatment with fungus, P. sordida SK7 was the most effective for enhancing enzymatic hydrolysis, approximately, 40% improvement.

Keywords: Corncob, Enzymatic hydrolysis, Microorganisms, Pretreatment.

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46 Statistical Optimization of Enzymatic Hydrolysis of Potato (Solanum tuberosum) Starch by Immobilized α-amylase

Authors: N.Peatciyammal, B.Balachandar, M.Dinesh Kumar, K.Tamilarasan, C.Muthukumaran

Abstract:

Enzymatic hydrolysis of starch from natural sources finds potential application in commercial production of alcoholic beverage and bioethanol. In this study the effect of starch concentration, temperature, time and enzyme concentration were studied and optimized for hydrolysis of Potato starch powder (of mesh 80/120) into glucose syrup by immobilized (using Sodium arginate) α-amylase using central composite design. The experimental result on enzymatic hydrolysis of Potato starch was subjected to multiple linear regression analysis using MINITAB 14 software. Positive linear effect of starch concentration, enzyme concentration and time was observed on hydrolysis of Potato starch by α-amylase. The statistical significance of the model was validated by F-test for analysis of variance (p ≤ 0.01). The optimum value of starch concentration, enzyme concentration, temperature, time and were found to be 6% (w/v), 2% (w/v), 40°C and 80min respectively. The maximum glucose yield at optimum condition was 2.34 mg/mL.

Keywords: Alcoholic beverage, Central Composite Design, Enzymatic hydrolysis, Glucose yield, Potato Starch.

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

Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej

Abstract:

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.

Keywords: Sugarcane bagasse, Microorganisms, Pretreatment, Enzymatic hydrolysis.

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44 New Kinetic Approach to the Enzymatic Hydrolysis of Proteins – A Case of Thermolysin-Catalyzed Albumin

Authors: Anna Trusek-Holownia, Andrzej Noworyta

Abstract:

Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: Peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides.

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

Authors: Md. Z. Alam, Amal A. Elgharbawy, Muhammad Moniruzzaman, Nassereldeen A. Kabbashi, Parveen Jamal

Abstract:

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.

Keywords: Cellulase, hydrolysis, lignocellulose, pretreatment, stability.

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42 Enhancing Efficiency for Reducing Sugar from Cassava Bagasse by Pretreatment

Authors: S. Gaewchingduang, P. Pengthemkeerati

Abstract:

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.

Keywords: Acid hydrolysis, cassava bagasse, enzymatic hydrolysis, hydrothermal pretreatment.

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41 Hydrolysis of Hull-Less Pumpkin Oil Cake Protein Isolate by Pepsin

Authors: Ivan Živanović, Žužana Vaštag, Senka Popović, Ljiljana Popović, Draginja Peričin

Abstract:

The present work represents an investigation of the hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein isolate (PuOC PI) by pepsin. To examine the effectiveness and suitability of pepsin towards PuOC PI the kinetic parameters for pepsin on PuOC PI were determined and then, the hydrolysis process was studied using Response Surface Methodology (RSM). The hydrolysis was carried out at temperature of 30°C and pH 3.00. Time and initial enzyme/substrate ratio (E/S) at three levels were selected as the independent parameters. The degree of hydrolysis, DH, was mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3 mA/mg proteins. Since the proposed second-order polynomial model showed good fit with the experimental data (R2 = 0.9822), the obtained mathematical model could be used for monitoring the hydrolysis of PuOC PI by pepsin, under studied experimental conditions, varying the time and initial E/S. To achieve the highest value of DH (39.13 %), the obtained optimum conditions for time and initial E/S were 30 min and 1.024 mA/mg proteins.

Keywords: Enzymatic hydrolysis, Pepsin, Pumpkin (CucurbitaPepo L.) oil cake protein isolate, Response surface methodology.

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40 Thermal and Morphological Evaluation of Chemically Pretreated Sugarcane Bagasse

Authors: Glauber Cruz, Patrícia A. S. Monteiro, Carlos E. M. Braz, Paulo Seleghin Jr., Igor Polikarpov, Paula M.Crnkovic

Abstract:

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.

Keywords: Alkaline solutions, bioethanol production, dilute acid, enzymatic hydrolysis, lignocellulosic biomass.

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39 Physical Properties and Resistant Starch Content of Rice Flour Residues Hydrolyzed by α-Amylase

Authors: Waranya Pongpaiboon, Warangkana Srichamnong, Supat Chaiyakul

Abstract:

Enzymatic modification of rice flour can produce highly functional derivatives use in food industries. This study aimed to evaluate the physical properties and resistant starch content of rice flour residues hydrolyzed by α-amylase. Rice flour hydrolyzed by α-amylase (60 and 300 u/g) for 1, 24 and 48 hours were investigated. Increasing enzyme concentration and hydrolysis time resulted in decreased rice flour residue’s lightness (L*) but increased redness (a*) and yellowness (b*) of rice flour residues. The resistant starch content and peak viscosity increased when hydrolysis time increased. Pasting temperature, trough viscosity, breakdown, final viscosity, setback and peak time of the hydrolyzed flours were not significantly different (p>0.05). The morphology of native flour was smooth without observable pores and polygonal with sharp angles and edges. However, after hydrolysis, granules with a slightly rough and porous surface were observed and a rough and porous surface was increased with increasing hydrolyzed time. The X-ray diffraction patterns of native flour showed A-type configuration, which hydrolyzed flour showed almost 0% crystallinity indicated that both amorphous and crystalline structures of starch were simultaneously hydrolyzed by α-amylase.

Keywords: α-Amylase, Enzymatic hydrolysis, Pasting properties, Resistant starch

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38 Characterization of Corn Cobs from Microwave and Potassium Hydroxide Pretreatment

Authors: Boonyisa Wanitwattanarumlug, Apanee Luengnaruemitchai, Sujitra Wongkasemjit

Abstract:

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.

Keywords: Corn cobs, Enzymatic hydrolysis, Microwave, Potassium hydroxide, Pretreatment.

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37 Effects of Li2O Thickness and Moisture Content on LiH Hydrolysis Kinetics in Slightly Humidified Argon

Authors: S. Xiao, M. B. Shuai, M. F. Chu

Abstract:

The hydrolysis kinetics of polycrystalline lithium hydride (LiH) in argon at various low humidities was measured by gravimetry and Raman spectroscopy with ambient water concentration ranging from 200 to 1200 ppm. The results showed that LiH hydrolysis curve revealed a paralinear shape, which was attributed to two different reaction stages that forming different products as explained by the 'Layer Diffusion Control' model. Based on the model, a novel two-stage rate equation for LiH hydrolysis reactions was developed and used to fit the experimental data for determination of Li2O steady thickness Hs and the ultimate hydrolysis rate vs. The fitted data presented a rise of Hs as ambient water concentration cw increased. However, in spite of the negative effect imposed by Hs increasing, the upward trend of vs remained, which implied that water concentration, rather than Li2O thickness, played a predominant role in LiH hydrolysis kinetics. In addition, the proportional relationship between vsHs and cw predicted by rate equation and confirmed by gravimetric data validated the model in such conditions.

Keywords: Hydrolysis kinetics, ‘Layer Diffusion Control’ model, Lithium hydride

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36 Hydrolysis Characteristics of Polycrystalline Lithium Hydride Powders and Sintered Bulk

Authors: M. B. Shuai, S. Xiao, Q. S. Li, M. F. Chu, X. F. Yang

Abstract:

Ambient hydrolysis products in moist air and hydrolysis kinetics in argon with humidity of RH1.5% for polycrystalline LiH powders and sintered bulks were investigated by X-ray diffraction, Raman spectroscopy and gravimetry. The results showed that the hydrolysis products made up a layered structure of LiOH•H2O/LiOH/Li2O from surface of the sample to inside. In low humid argon atmosphere, the primary hydrolysis product was Li2O rather than LiOH. The hydrolysis kinetic curves of LiH bulks present a paralinear shape, which could be explained by the “Layer Diffusion Control" model. While a three-stage hydrolysis kinetic profile was observed for LiH powders under the same experimental conditions. The first two sections were similar to that of the bulk samples, and the third section also presents a linear reaction kinetics but with a smaller reaction rate compared to the second section because of a larger exothermic effect for the hydrolysis reaction of LiH powder.

Keywords: Hydrolysis, lithium compound, polycrystallinelithium hydride

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35 Enzymatic Synthesis of Olive-Based Ferulate Esters: Optimization by Response Surface Methodology

Authors: S. Mat Radzi, N. J. Abd Rahman, H. Mohd Noor, N. Ariffin

Abstract:

Ferulic acid has widespread industrial potential by virtue of its antioxidant properties. However, it is partially soluble in aqueous media, limiting their usefulness in oil-based processes in food, cosmetic, pharmaceutical, and material industry. Therefore, modification of ferulic acid should be made by producing of more lipophilic derivatives. In this study, a preliminary investigation of lipase-catalyzed trans-esterification reaction of ethyl ferulate and olive oil was investigated. The reaction was catalyzed by immobilized lipase from Candida antarctica (Novozym 435), to produce ferulate ester, a sunscreen agent. A statistical approach of Response surface methodology (RSM) was used to evaluate the interactive effects of reaction temperature (40-80°C), reaction time (4-12 hours), and amount of enzyme (0.1-0.5 g). The optimum conditions derived via RSM were reaction temperature 60°C, reaction time 2.34 hours, and amount of enzyme 0.3 g. The actual experimental yield was 59.6% ferulate ester under optimum condition, which compared well to the maximum predicted value of 58.0%.

Keywords: Ferulic acid, Enzymatic Synthesis, Esters, RSM.

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34 Enzymatic Activity of Alfalfa in a Phenanthrene-contaminated Environment

Authors: Anna Yu. Muratova, Vera V. Kapitonova, Marina P. Chernyshova, Olga V. Turkovskaya

Abstract:

This research was undertaken to study enzymatic activity in the shoots, roots, and rhizosphere of alfalfa (Medicago sativa L.) grown in quartz sand that was uncontaminated and contaminated with phenanthrene at concentrations of 10 and 100 mg kg-1. The higher concentration of phehanthrene had a distinct phytotoxic effect on alfalfa, inhibiting seed germination energy, plant survival, and biomass accumulation. The plant stress response to the environmental pollution was an increase in peroxidase activity. Peroxidases were the predominant enzymes in the alfalfa shoots and roots. The peroxidase profile in the shoots differed from that in the roots and had different isoenzyme numbers. 2,2'-Azinobis-(3-ethylbenzo-thiazoline-6-sulphonate) (ABTS) peroxidase was predominant in the shoots, and 2,7-diaminofluorene (2,2-DAF) peroxidase was predominant in the roots. Under the influence of phenanthrene, the activity of 2,7-DAF peroxidase increased in the shoots, and the activity of ABTS peroxidase increased in the roots. Alfalfa root peroxidases were the prevalent enzyme systems in the rhizosphere sand. Examination of the activity of alfalfa root peroxidase toward phenanthrene revealed the possibility of involvement of the plant enzyme in rhizosphere degradation of the PAH.

Keywords: Medicago sativa, enzymatic activity, peroxidase, phenanthrene.

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33 Production of Bioethanol through Hydrolysis of Agro-Industrial Banana Crop Residues

Authors: Sánchez Acuña, Juan Camilo, Granados Gómez, Mildred Magaly, Navarrete Rodríguez, Luisa Fernanda

Abstract:

Nowadays, the main biofuels source production as bioethanol is food crops. This means a high competition between foods and energy production. For this reason, it is necessary to take into account the use of new raw materials friendly to the environment. The main objective of this paper is to evaluate the potential of the agro-industrial banana crop residues in the production of bioethanol. A factorial design of 24 was used, the design has variables such as pH, time and concentration of hydrolysis, another variable is the time of fermentation that is of 7 or 15 days. In the hydrolysis phase, the pH is acidic (H2SO4) or basic (NaOH), the time is 30 or 15 minutes and the concentration is 0.1 or 0.5 M. It was observed that basic media, low concentrations, fermentation, and higher pretreatment times produced better performance in terms of biofuel obtained.

Keywords: Bioethanol, biofuels, banana waste, hydrolysis.

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32 Production of Glucose from the Hydrolysis of Cassava Residue using Bacteria Isolates from Thai Higher Termites

Authors: Pitcha Wongskeo, Pramoch Rangsunvigit, Sumaeth Chavadej

Abstract:

The possibility of using cassava residue containing 49.66% starch, 21.47% cellulose, 12.97% hemicellulose, and 21.86% lignin as a raw material to produce glucose using enzymatic hydrolysis was investigated. In the experiment, each reactor contained the cassava residue, bacteria cells, and production medium. The effects of particles size (40 mesh and 60 mesh) and strains of bacteria (A002 and M015) isolated from Thai higher termites, Microcerotermes sp., on the glucose concentration at 37°C were focused. High performance liquid chromatography (HPLC) with a refractive index detector was used to determine the quantity of glucose. The maximum glucose concentration obtained at 37°C using strain A002 and 60 mesh of the cassava residue was 1.51 g/L at 10 h.

Keywords: Hydrolysis, termites, glucose, cassava

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

Authors: T. R. Bandara, H. Jaelani, G. J. Griffin

Abstract:

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.

Keywords: Biomass, bagasse, hydrolysis, monosaccharide, supported liquid membrane, purification.

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30 Pseudo-Homogeneous Kinetic of Dilute-Acid Hydrolysis of Rice Huskfor Ethanol Production: Effect of Sugar Degradation

Authors: Megawati, Wahyudi B. Sediawan, Hary Sulistyo, Muslikhin Hidayat

Abstract:

Rice husk is a lignocellulosic source that can be converted to ethanol. Three hundreds grams of rice husk was mixed with 1 L of 0.18 N sulfuric acid solutions then was heated in an autoclave. The reaction was expected to be at constant temperature (isothermal), but before that temperature was achieved, reaction has occurred. The first liquid sample was taken at temperature of 140 0C and repeated every 5 minute interval. So the data obtained are in the regions of non-isothermal and isothermal. It was observed that the degradation has significant effects on the ethanol production. The kinetic constants can be expressed by Arrhenius equation with the frequency factors for hydrolysis and sugar degradation of 1.58 x 105 min-1 and 2.29 x 108 L/mole-min, respectively, while the activation energies are 64,350 J/mole and 76,571 J/mole. The highest ethanol concentration from fermentation is 1.13% v/v, attained at 220 0C.

Keywords: degradation, ethanol, hydrolysis, rice husk.

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29 Reducing Sugar Production from Durian Peel by Hydrochloric Acid Hydrolysis

Authors: Matura Unhasirikul, Nuanphan Naranong, Woatthichai Narkrugsa

Abstract:

Agricultural waste is mainly composed of cellulose and hemicelluloses which can be converted to sugars. The inexpensive reducing sugar from durian peel was obtained by hydrolysis with HCl concentration at 0.5-2.0% (v/v). The hydrolysis range of time was for 15-60 min when the mixture was autoclaved at 121 °C. The result showed that acid hydrolysis efficiency (AHE) highest to 80.99% at condition is 2.0%concentration for 15 min. Reducing sugar highest to 56.07 g/litre at condition is 2.0% concentration for 45min. Total sugar highest to 59.83 g/litre at condition is 2.0%concentration for 45min, which was not significant (p < 0.05) with condition 2.0% concentration for 30 min and 1.5 % concentration for 45 and 60 min. The increase in concentration increased AHE, reducing sugar and total sugar. The hydrolysis time had no effect on AHE, reducing sugar and total sugar. The maximum reducing sugars of each concentration were at hydrolysis time 45 min .The hydrolysated were analysis by HPLC, the results revealed that the principle of sugar were glucose, fructose and xylose.

Keywords: acid hydrolysis efficiency (AHE), reducing sugar, total sugar

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28 Hydrolysis of Eicchornia crassipes and Egeria densa for Ethanol Production by Yeasts Isolated from Colombian Lake Fúquene

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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27 Enzymatic Saccharification of Dilute Alkaline Pre-treated Microalgal (Tetraselmis suecica) Biomass for Biobutanol Production

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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26 Pseudo-Homogeneous Kinetic of Dilute-Acid Hydrolysis of Rice Husk for Ethanol Production: Effect of Sugar Degradation

Authors: Megawati, Wahyudi B. Sediawan, Hary Sulistyo, Muslikhin Hidayat

Abstract:

Rice husk is a lignocellulosic source that can be converted to ethanol. Three hundreds grams of rice husk was mixed with 1 L of 0.18 N sulfuric acid solutions then was heated in an autoclave. The reaction was expected to be at constant temperature (isothermal), but before that temperature was achieved, reaction has occurred. The first liquid sample was taken at temperature of 140 0C and repeated every 5 minute interval. So the data obtained are in the regions of non-isothermal and isothermal. It was observed that the degradation has significant effects on the ethanol production. The kinetic constants can be expressed by Arrhenius equation with the frequency factors for hydrolysis and sugar degradation of 1.58 x 105 1/min and 2.29 x 108 L/mole/min, respectively, while the activation energies are 64,350 J/mole and 76,571 J/mole. The highest ethanol concentration from fermentation is 1.13% v/v, attained at 220 0C.

Keywords: degradation, ethanol, hydrolysis, rice husk

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25 Utilization of Sugarcane Bagasses for Lactic Acid Production by acid Hydrolysis and Fermentation using Lactobacillus sp

Authors: Woranart Jonglertjunya, Nattawadee Pranrawang, Nuanyai Phookongka, Thanasak Sridangtip, Watthana Sawedrungreang, Chularat Krongtaew

Abstract:

Sugarcane bagasses are one of the most extensively used agricultural residues. Using acid hydrolysis and fermentation, conversion of sugarcane bagasses to lactic acid was technically and economically feasible. This research was concerned with the solubility of lignin in ammonium hydroxide, acid hydrolysis and lactic acid fermentation by Lactococcus lactis, Lactobacillus delbrueckii, Lactobacillus plantarum, and Lactobacillus casei. The lignin extraction results for different ammonium hydroxide concentrations showed that 10 % (v/v) NH4OH was favorable to lignin dissolution. Acid hydrolysis can be enhanced with increasing acid concentration and reaction temperature. The optimum glucose and xylose concentrations occurred at 121 ○C for 1 hour hydrolysis time in 10% sulphuric acid solution were 32 and 11 g/l, respectively. In order to investigate the significance of medium composition on lactic acid production, experiments were undertaken whereby a culture of Lactococcus lactis was grown under various glucose, peptone, yeast extract and xylose concentrations. The optimum medium was composed of 5 g/l glucose, 2.5 g/l xylose, 10 g/l peptone and 5 g/l yeast extract. Lactococcus lactis represents the most efficient for lactic acid production amongst those considered. The lactic acid fermentation by Lactococcus lactis after 72 hours gave the highest yield of 1.4 (g lactic acid per g reducing sugar).

Keywords: sugarcane bagasses, acid hydrolysis, lactic acid, fermentation

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24 Mathematical modeling of Bi-Substrate Enzymatic Reactions with Ping-Pong Mechanism in the Presence of Competitive Inhibitors

Authors: Rafayel A. Azizyan, Aram E. Gevogyan, Valeri B. Arakelyan, Emil S. Gevorgyan

Abstract:

The mathematical modeling of different biological processes is usually used to predict or assess behavior of systems in which these processes take place. This study deals with mathematical and computer modeling of bi-substrate enzymatic reactions with ping-pong mechanism, which play an important role in different biochemical pathways. Besides that, three models of competitive inhibition were designed using different software packages. The main objective of this study is to represent the results from in silico investigation of bi-substrate enzymatic reactions with ordered pingpong mechanism in the presence of competitive inhibitors, as well as to describe in details the inhibition effects. The simulation of the models with certain kinetic parameters allowed investigating the behavior of reactions as well as determined some interesting aspects concerning influence of different cases of competitive inhibition. Simultaneous presence of two inhibitors, competitive to the S1 and S2 substrates have been studied. Moreover, we have found the pattern of simultaneous influence of both inhibitors.

Keywords: Mathematical modeling, bi-substrate enzymatic reactions, ping-pong mechanism, competitive inhibition.

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23 Mathematical Modeling of Uncompetitive Inhibition of Bi-Substrate Enzymatic Reactions

Authors: Rafayel A. Azizyan, Aram E. Gevorgyan, Valeri B. Arakelyan, Emil S. Gevorgyan

Abstract:

Currently, mathematical and computer modeling are widely used in different biological studies to predict or assess behavior of such a complex systems as a biological are. This study deals with mathematical and computer modeling of bi-substrate enzymatic reactions, which play an important role in different biochemical pathways. The main objective of this study is to represent the results from in silico investigation of bi-substrate enzymatic reactions in the presence of uncompetitive inhibitors, as well as to describe in details the inhibition effects. Four models of uncompetitive inhibition were designed using different software packages. Particularly, uncompetitive inhibitor to the first [ES1] and the second ([ES1S2]; [FS2]) enzyme-substrate complexes have been studied. The simulation, using the same kinetic parameters for all models allowed investigating the behavior of reactions as well as determined some interesting aspects concerning influence of different cases of uncompetitive inhibition. Besides, it has been shown that uncompetitive inhibitors exhibit specific selectivity depending on mechanism of bi-substrate enzymatic reaction. 

Keywords: Mathematical modeling, bi-substrate enzymatic reactions, sequential mechanism, ping-pong mechanism, uncompetitive inhibition.

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22 Degree of Hydrolysis of Proteinaceous Components of Porang Flour Using Papain

Authors: Fadilah Fadilah, Rochmadi Rochmadi, Siti Syamsiah, Djagal W. Marseno

Abstract:

Glucomannan can be found in the tuber of porang together with starch and proteinaceous components which were regarded as impurities. An enzymatic process for obtaining higher glucomannan content from Porang flour have been conducted. Papain was used for hydrolysing proteinaceous components in Porang flour which was conducted after a simultaneous extraction of glucomannan and enzymatic starch hydrolysis. Three variables affecting the rate were studied, i.e. temperature, the amount of enzyme and the stirring speed. The ninhydrin method was used to determine degree of protein hydrolysis. Results showed that the rising of degree of hydrolysis were fast in the first ten minutes of the reaction and then proceeded slowly afterward. The optimum temperature for hydrolysis was 60 oC. Increasing the amount of enzyme showed a remarkable effect to degree of hydrolysis, but the stirring speed had no significant effect. This indicated that the reaction controlled the rate of hydrolysis.

Keywords: Degree of hydrolysis, ninhydrin, papain, porang flour, proteinaceous components.

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21 The Contribution of the PCR-Enzymatic Digestion in the Positive Diagnosis of Proximal Spinal Muscular Atrophy in the Moroccan Population

Authors: H. Merhni, A. Sbiti, I. Ratbi, A. Sefiani

Abstract:

The proximal spinal muscular atrophy (SMA) is a group of neuromuscular disorders characterized by progressive muscle weakness due to the degeneration and loss of anterior motor neurons of the spinal cord. Depending on the age of onset of symptoms and their evolution, four types of SMA, varying in severity, result in a mutations of the SMN gene (survival of Motor neuron). We have analyzed the DNA of 295 patients referred to our genetic counseling; since January 1996 until October 2014; for suspected SMA. The homozygous deletion of exon 7 of the SMN gene was found in 133 patients; of which, 40.6% were born to consanguineous parents. In countries like Morocco, where the frequency of heterozygotes for SMA is high, genetic testing should be offered as first-line and, after careful clinical assessment, especially in newborns and infants with congenital hypotonia unexplained and prognosis compromise. The molecular diagnosis of SMA allows a quick and certainly diagnosis, provide adequate genetic counseling for families at risk and suggest, for couples who want prenatal diagnosis. The analysis of the SMN gene is a perfect example of genetic testing with an excellent cost/benefit ratio that can be of great interest in public health, especially in low-income countries. We emphasize in this work for the benefit of the generalization of molecular diagnosis of SMA by the technique of PCR-enzymatic digestion in other centers in Morocco.

Keywords: Exon7, PCR-digestion, SMA, SMN gene.

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20 Qualitative Characteristics of Meat from Lambs Fed Hydrolyzed Sugarcane

Authors: V. Endo, A. G. Silva Sobrinho, F. A. Almeida, N. L. L. Lima, G. M. Manzi, L. G. A. Cirne, N. M. B. L. Zeola

Abstract:

We used 24 Ile de France lambs, weighing between 15 and 32 kg (BW). Treatments were supplemented with concentrate: “in nature” sugarcane (IN), sugarcane hydrolyzed using 0.6% calcium oxide (CaO) under aerobic condition (AER), and sugarcane hydrolyzed using 0.6% CaO under anaerobic condition (ANA), constituting a completely randomized design with eight repetitions per treatment. Lambs were housed in individual stalls and fed into the through, allowing 10% of leftovers. Lambs were slaughtered when body weight reached 32 kg. The following parameters were determined on Longissimu lumborum muscle of hot and cold carcasses: pH and color, 45 minutes and 24 hours after slaughtering. Qualitative analysis of the meat were performed in the loins, water-holding capacity (WHC), cooking loss (CL), and shear force (SF). We used a completely randomized design with three treatments and eight repetitions. Means were compared by Tukey test at 5% significance. A higher value for redness (a*) 45 minutes after slaughter (10.48) were found for lambs fed hydrolyzed under anaerobic conditions sugarcane. The other qualitative characteristics of meat were not affected by treatments (P >0.05). The comparison of meat quality resulting from the treatments shows that it is possible to feed in nature sugarcane to lambs, thus waiving hydrolyses process and the spending with alkalizing agent.

Keywords: Calcium oxide, hydrolysis, meat quality, pH.

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