Search results for: enzyme kinetics

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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Authors: Fatima Zohra Ibn Majdoub Hassani, Ivan Lavandera, Joseph Kreit

Abstract:

This study explored the effects of different organic solvents, temperature, and the amount of glycerol on the alcohol dehydrogenase (ADH)-catalysed stereoselective reduction of different ketones. These conversions were then analyzed by gas chromatography. It was found that when the amount of deep eutectic solvents (DES) increases, it can improve the stereoselectivity of the enzyme although reducing its ability to convert the substrate into the corresponding alcohol. Moreover, glycerol was found to have a strong stabilizing effect on the ADH from Ralstonia sp. (E. coli/ RasADH). In the case of organic solvents, it was observed that the best conversions into the alcohols were achieved with DMSO and hexane. It was also observed that temperature decreased the ability of the enzyme to convert the substrates into the products and also affected the selectivity. In addition to that, the recycling of DES up to three times gave good conversions and enantiomeric excess results and glycerol showed a positive effect in the stability of various ADHs. Using RasADH, a good conversion and enantiomeric excess into the S-alcohol were obtained. It was found that an enhancement of the temperature disabled the stabilizing effect of glycerol and decreased the stereoselectivity of the enzyme. However, for other ADHs a temperature increase had an opposite positive effect, especially with ADH-T from Thermoanaerobium sp. One of the objectives of this study was to see the effect of cofactors such as NAD(P) on the biocatlysis activities of ADHs.

Keywords: Alcohol dehydrogenases, DES, gas chromatography, RasADH.

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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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Authors: Wen-Shyong Tzou, Chih-Ching Huang, Chin-Hwa Hu, Ying-Tsang Lo, Tun-Wen Pai, Chia-Yin Chiang, Chung-Hao Li, Hong-Jyuan Jian

Abstract:

Plasmin plays an important role in the human circulatory system owing to its catalytic ability of fibrinolysis. The immediate injection of plasmin in patients of strokes has intrigued many scientists to design vectors that can transport plasmin to the desired location in human body. Here we predict the structure of human plasmin and investigate the interaction of plasmin with the gold-nanoparticle. Because the crystal structure of plasminogen has been solved, we deleted N-terminal domain (Pan-apple domain) of plasminogen and generate a mimic of the active form of this enzyme (plasmin). We conducted a simulated annealing process on plasmin and discovered a very large conformation occurs. Kringle domains 1, 4 and 5 had been observed to leave its original location relative to the main body of the enzyme and the original doughnut shape of this enzyme has been transformed to a V-shaped by opening its two arms. This observation of conformational change is consistent with the experimental results of neutron scattering and centrifugation. We subsequently docked the plasmin on the simulated gold surface to predict their interaction. The V-shaped plasmin could utilize its Kringle domain and catalytic domain to contact the gold surface. Our findings not only reveal the flexibility of plasmin structure but also provide a guide for the design of a plasmin-gold nanoparticle.

Keywords: Docking, gold nanoparticle, molecular simulation, plasmin.

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Authors: Ratchadaporn Oonsivilai, Jutarat Manatwiyangkool, Anant Oonsivilai

Abstract:

Theoptimal extraction condition of dried Phaseolus vulgaris powderwas studied. The three independent variables are raw material concentration, shaking and centrifugaltime. The dependent variables are both yield percentage of crude extract and alphaamylase enzyme inhibition activity. The experimental design was based on box-behnkendesign. Highest yield percentage of crude extract could get from extraction condition at concentration of 1, 0,1, concentration of 0.15 M ,extraction time for 2hour, and separationtime for60 min. Moreover, the crude extract with highest alpha-amylase enzyme inhibition activityoccurred by extraction condition at concentration of 0.10 M, extraction time for 2 min, and separation time for 45 min

Keywords: Extraction time, Optimal condition, Alpha-amylase enzymeinhibition activity

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Authors: Turan Çalban, Nursel Keskin, Sabri Çolak, Soner Kuşlu

Abstract:

Gypsum (CaSO4.2H2O) is a mineral that is found in large quantities in the Turkey and in the World. In this study, the dissolution of this mineral in the diammonium hydrogen phosphate solutions has been studied. The dissolution and dissolution kinetics of gypsum in diammonium hydrogen phosphate solutions will be useful for evaluating of solid wastes containing gypsum. Parameters such as diammonium hydrogen phosphate concentration, temperature and stirring speed affecting on the dissolution rate of the gypsum in diammonium hydrogen phosphate solutions were investigated. In experimental studies have researched effectiveness of the selected parameters. The dissolution of gypsum were examined in two parts at low and high temperatures. The experimental results were successfully correlated by linear regression using Statistica program. Dissolution curves were evaluated shrinking core models for solidfluid systems. The activation energy was found to be 34.58 kJ/mol and 44.45 kJ/mol for the low and the high temperatures. The dissolution of gypsum was controlled by chemical reaction both low temperatures and high temperatures.

Keywords: Diammonium hydrogen phosphate, Dissolution, Gypsum, Kinetics.

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Authors: A. Peter Amala Sujith, T.V. Hymavathi, P. Yasoda Devi

Abstract:

Lutein is a dietary oxycarotenoid which is found to reduce the risks of Age-related Macular Degeneration (AMD). Supercritical fluid extraction of lutein esters from marigold petals was carried out and was found to be much effective than conventional solvent extraction. The saponification of pre-concentrated lutein esters to produce free lutein was studied which showed a composition of about 88% total carotenoids (UV-VIS spectrophotometry) and 90.7% lutein (HPLC). The lipase catalyzed hydrolysis of lutein esters in conventional medium was investigated. The optimal temperature, pH, enzyme concentration and water activity were found to be 50°C, 7, 15% and 0.33 respectively and the activity loss of lipase was about 25% after 8 times re-use in at 50°C for 12 days. However, the lipase catalyzed hydrolysis of lutein esters in conventional media resulted in poor conversions (16.4%).

Keywords: lutein, preconcentration, saponification, lipase

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Authors: Aparajita Goswami, Mihir K Purkait

Abstract:

In the present study Schwertmannite (an iron oxide hydroxide) is selected as an adsorbent for defluoridation of water. The adsorbent was prepared by wet chemical process and was characterized by SEM, XRD and BET. The fluoride adsorption efficiency of the prepared adsorbent was determined with respect to contact time, initial fluoride concentration, adsorbent dose and pH of the solution. The batch adsorption data revealed that the fluoride adsorption efficiency was highly influenced by the studied factors. Equilibrium was attained within one hour of contact time indicating fast kinetics and the adsorption data followed pseudo second order kinetic model. Equilibrium isotherm data fitted to both Langmuir and Freundlich isotherm models for a concentration range of 5-30 mg/L. The adsorption system followed Langmuir isotherm model with maximum adsorption capacity of 11.3 mg/g. The high adsorption capacity of Schwertmannite points towards the potential of this adsorbent for fluoride removal from aqueous medium.

Keywords: Adsorption, fluoride, isotherm study, kinetics, schwertmannite.

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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: Norzita Ngadi, John Abrahamson, Conan Fee, Ken Morison

Abstract:

Nonspecific protein adsorption generally occurs on any solid surfaces and usually has adverse consequences. Adsorption of proteins onto a solid surface is believed to be the initial and controlling step in biofouling. Surfaces modified with end-tethered poly(ethylene glycol) (PEG) have been shown to be protein-resistant to some degree. In this study, the adsorption of β-casein and lysozyme was performed on 6 different types of surfaces where PEG was tethered onto stainless steel by polyethylene imine (PEI) through either OH or NHS end groups. Protein adsorption was also performed on the bare stainless steel surface as a control. The adsorption was conducted at 23 °C and pH 7.2. In situ QCM-D was used to determine PEG adsorption kinetics, plateau PEG chain densities, protein adsorption kinetics and plateau protein adsorbed quantities. PEG grafting density was the highest for a NHS coupled chain, around 0.5 chains / nm2. Interestingly, lysozyme which has smaller size than β-casein, appeared to adsorb much less mass than that of β- casein. Overall, the surface with high PEG grafting density exhibited a good protein rejection.

Keywords: QCM-D, PEG, stainless steel, β-casein, lysozyme.

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Authors: Pavlo Selyshchev, Samuel Akintunde

Abstract:

A theoretical approach to consider formation of chemical compound layer at the interface between initial substances A and B due to the interfacial interaction and diffusion is developed. It is considered situation when speed of interfacial interaction is large enough and diffusion of A-atoms through AB-layer is much more then diffusion of B-atoms. Atoms from A-layer diffuse toward B-atoms and form AB-atoms on the surface of B-layer. B-atoms are assumed to be immobile. The growth kinetics of the AB-layer is described by two differential equations with non-linear coupling, producing a good fit to the experimental data. It is shown that growth of the thickness of the AB-layer determines by dependence of chemical reaction rate on reactants concentration. In special case the thickness of the AB-layer can grow linearly or parabolically depending on that which of processes (interaction or the diffusion) controls the growth. The thickness of AB-layer as function of time is obtained. The moment of time (transition point) at which the linear growth are changed by parabolic is found.

Keywords: Phase formation, Binary systems, Interfacial Reaction, Diffusion, Compound layers, Growth kinetics.

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Authors: Zhen Chen, Haitao Zhang, Weiyong Ying, Dingye Fang

Abstract:

Based on a global kinetics of direct dimethyl ether (DME) synthesis process from syngas, a steady-state one-dimensional mathematical model for the bubble column slurry reactor (BCSR) has been established. It was built on the assumption of plug flow of gas phase, sedimentation-dispersion model of catalyst grains and isothermal chamber regardless of reaction heats and rates for the design of an industrial scale bubble column slurry reactor. The simulation results indicate that higher pressure and lower temperature were favorable to the increase of CO conversion, DME selectivity, products yield and the height of slurry bed, which has a coincidence with the characteristic of DME synthesis reaction system, and that the height of slurry bed is lessen with the increasing of operation temperature in the range of 220-260℃. CO conversion, the optimal operation conditions in BCSR were proposed. 

Keywords: Alcohol/ether fuel, bubble column slurry reactor, global kinetics, mathematical model.

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Authors: Soni Yadav, Sitansu Kumar Verma, Jitendra Singh, Ajay Kumar

Abstract:

This article comprises detail information about L-asparaginase, encompassing topic such as various sources of L-asparaginase, mechanism and properties of L-asparaginase. Also describe the production, cultivation and purification of L-asparaginase along with information about the application of L-asparaginase. L-asparaginase catalyzes the conversion reaction to convert asparagine to aspartic acid and ammonia. Asparagine is a nutritional requirement for both normal and tumor cell. Present scenario has found that L-asparaginase has been found to be a best anti tumor or antileukemic agent. In the recent years this enzyme gained application in the field of clinical research pharmacologic and food industry. It has been characterized based on the enzyme assay principle hydrolyzing L-asparagine into L-aspartic acid and ammonia. It has been observed that eukaryotic microorganisms such as yeast and filamentous fungi have a potential for L-asparaginase production. L-asparaginase has been and is still one of the most lengthily studied therapeutic enzymes by scientist and researchers worldwide.

Keywords: L-asparaginase, antitumor, solid state fermentation, chemotherapeutic.

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Authors: F. Bamdad, Sh. Dokhani, J. Keramat, R. Zareie

Abstract:

Biologically active peptides are of particular interest in food science and human nutrition because they have been shown to play several physiological roles. In vitro gastrointestinal digestion of lentil and whey proteins in this study produced high angiotensin-I converting enzyme inhibitory activity with 75.5±1.9 and 91.4±2.3% inhibition, respectively. High ACE inhibitory activity was observed in lentil after 5 days of germination (84.3±1.2%). Fractionation by reverse phase chromatography gave inhibitory activities as high as 86.3±2.0 for lentil, 94.8±1.8% for whey and 93.7±1.7% at 5th day of germination. Further purification by HPLC resulted in several inhibitory peptides with IC50 values ranging from 0.064 to 0.164 mg/ml. These results demonstrate that lentil proteins are a good source of peptides with ACE inhibitory activity that can be released by germination or gastrointestinal digestion. Despite the lower bioactivity in comparison with whey proteins, incorporation of lentil proteins in functional food formulations and natural drugs look promising.

Keywords: ACE inhibitory peptides, digestion, germination, lentil proteins, whey proteins

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Authors: Le Thuy Mai, Vu Nguyen Thanh

Abstract:

β-Glucosidase is an important enzyme for production of ethanol from lignocellulose. With hydrolytic activity on cellooligosaccharides, especially cellobiose, β-glucosidase removes product inhibitory effect on cellulases and forms fermentable sugars. In this study, β-glucosidase encoding gene (BGL1) from traditional starter yeast Saccharomycosis fibuligera BMQ908 was cloned and expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared 98% nucleotide homology with the closest GenBank sequence (M22475) but identity in amino-acid sequences of catalytic domains. Recombinant plasmid pPICZαA/BGL1 containing the sequence encoding BGL1 mature protein and α-factor secretion signal was constructed and transformed into methylotrophic yeast P. pastoris by electroporation. The recombinant strain produced single extracellular protein with molecular weight of 120 kDa and cellobiase activity of 60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0 and the optimum temperature was 50°C.

Keywords: β-Glucosidase, Pichia pastoris, Saccharomycopsisfibuligera, recombinant enzyme.

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Authors: A. Taoufyq, B. Bakiz, A. Benlhachemi, L. Patout, D. V. Chokouadeua, F. Guinneton, G. Nolibe, A. Lyoussi, J-R. Gavarri

Abstract:

The aim of this study was to investigate the photocatalytic activity of polycrystalline phases of bismuth tungstate of formula Bi2WO6. Polycrystalline samples were elaborated using a coprecipitation technique followed by a calcination process at different temperatures (300, 400, 600 and 900°C). The obtained polycrystalline phases have been characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Crystal cell parameters and cell volume depend on elaboration temperature. High-resolution electron microscopy images and image simulations, associated with X-ray diffraction data, allowed confirming the lattices and space groups Pca21. The photocatalytic activity of the as-prepared samples was studied by irradiating aqueous solutions of Rhodamine B, associated with Bi2WO6 additives having variable crystallite sizes. The photocatalytic activity of such bismuth tungstates increased as the crystallite sizes decreased. The high specific area of the photocatalytic particles obtained at 300°C seems to condition the degradation kinetics of RhB.

Keywords: Bismuth tungstate, crystallite sizes, electron microscopy, photocatalytic activity, X-ray diffraction.

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Authors: S. Mat Radzi, W.A.F. Mustafa, S.S Othman, H.M. Noor

Abstract:

Nowadays, butyl acetate, a pineapple flavor has been applied widely in food, beverage, cosmetic and pharmaceutical industries. In this study, Butyl acetate, a flavor ester was successfully synthesized via green synthesis of enzymatic reaction route. Commercial immobilized lipase from Rhizomucor miehei (Lipozyme RMIM) was used as biocatalyst in the esterification reaction between acetic acid and butanol. Various reaction parameters such as reaction time (RT), temperature (T) and amount of enzyme (E) were chosen to optimize the reaction synthesis in solvent-free system. The optimum condition to produce butyl acetate was at reaction time (RT), 18 hours; temperature (T), 37°C and amount of enzyme, 25 % (w/w of total substrate). Analysis of yield showed that at optimum condition, >78 % of butyl acetate was produced. The product was confirmed as butyl acetate from FTIR analysis whereby the presence of an ester group was observed at wavenumber of 1742 cm-1.

Keywords: Butyl acetate, immobilized enyzme, esterification, flavor ester, green synthesis

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Authors: Makhlouf Mourad, Messabih Sidi Mohamed, Bouchher Omar, Houali Farida, Benrachedi Khaled

Abstract:

Mesoporous materials are very commonly used as adsorbent materials for removing phenolic compounds. However, the adsorption mechanism of these compounds is still poorly controlled. However, understanding the interactions mesoporous materials/adsorbed molecules is very important in order to optimize the processes of liquid phase adsorption. The difficulty of synthesis is to keep an orderly and cubic pore structure and achieve a homogeneous surface modification. The grafting of Si(CH₃)₃ was chosen, to transform hydrophilic surfaces hydrophobic surfaces. The aim of this work is to study the kinetics and thermodynamics of two volatile organic compounds VOC phenol (PhOH) and P hydroxy benzoic acid (4AHB) on a mesoporous material of type MCM-48 grafted with an organosilane of the Trimethylchlorosilane (TMCS) type, the material thus grafted or functionalized (hereinafter referred to as MCM-48-G). In a first step, the kinetic and thermodynamic study of the adsorption isotherms of each of the VOCs in mono-solution was carried out. In a second step, a similar study was carried out on a mixture of these two compounds. Kinetic models (pseudo-first order, pseudo-second order) were used to determine kinetic adsorption parameters. The thermodynamic parameters of the adsorption isotherms were determined by the adsorption models (Langmuir, Freundlich). The comparative study of adsorption of PhOH and 4AHB proved that MCM-48-G had a high adsorption capacity for PhOH and 4AHB; this may be related to the hydrophobicity created by the organic function of TMCS in MCM-48-G. The adsorption results for the two compounds using the Freundlich and Langmuir models show that the adsorption of 4AHB was higher than PhOH. The values ​​obtained by the adsorption thermodynamics show that the adsorption interactions for our sample with the phenol and 4AHB are of a physical nature. The adsorption of our VOCs on the MCM-48 (G) is a spontaneous and exothermic process.

Keywords: Adsorption, kinetics, isotherm, mesoporous materials, TMCS, phenol, P-hydroxy benzoic acid.

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Authors: Abdul Rauf, Muhammad Irfan, Muhammad Nadeem, Ishtiaq Ahmed, Hafiz Muhammad Nasir Iqbal

Abstract:

Rhizopus oligosporus was used in the present study for the production of protease enzyme under SSF. Sunflower meal was used as by-product of oil industry incorporated with organic salts was employed for the production of protease enzyme. The main purpose of the present was to study different parameters of protease productivity, its yields and to optimize basal fermentation conditions. The optimal conditions found for protease production using sunflower meal as a substrate in the present study were inoculum size (1%), substrate (Sunflower meal), substrate concentration (20 g), pH (3), cultivation period (72 h), incubation temperature (35oC), substrate to diluent-s ratio (1:2) and tween 81 (1 mL). The maximum production of protease in the presence of cheaper substrate at low concentration and stability at acidic pH, these characteristics make the strain and its enzymes useful in different industry.

Keywords: Acidic protease, Rhizopus oligosporus, Mediaoptimization, Solid state Fermentation

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Authors: A. Magesh, B. Preetha, T. Viruthagiri

Abstract:

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.

Keywords: Fusarium oxysporum, Lignocellulosic biomass, Product formation kinetics, Statistical experimental design

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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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Authors: Mohamed H. Khalifa, Fikry I. El-Shahawi, Nabil A. Mansour

Abstract:

The cotton leafworm, Spodoptera littoralis (Boisduval) is a major insect pest of vegetables and cotton crops in Egypt, and exhibits different levels of tolerance to certain insecticides. Chlorantraniliprole has been registered recently in Egypt for control this insect. The susceptibilities of three S. littoralis populations collected from El Behaira governorate, north Egypt to chlorantraniliprole were determined by leaf-dipping technique on 4^th instar larvae. Obvious variation of toxicity was observed among the laboratory susceptible, and three field populations with LC₅₀ values ranged between 1.53 µg/ml and 6.22 µg/ml. However, all the three field populations were less susceptible to chlorantraniliprole than a laboratory susceptible population. The most tolerant populations were sampled from El Delengat (ED) Province where S. littoralis had been frequently challenged by insecticides. Certain enzyme activity assays were carried out to be correlated with the mechanism of the observed field population tolerance. All field populations showed significantly enhanced activities of detoxification enzymes compared with the susceptible strain. The regression analysis between chlorantraniliprole toxicities and enzyme activities revealed that the highest correlation is between α-esterase or β-esterase (α-β-EST) activity and collected field strains susceptibility, otherwise this correlation is not significant (P > 0.05). Synergism assays showed the ED and susceptible strains could be synergized by known detoxification inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.01-8.76-fold and 1.09-2.94 fold, respectively), TPP showed the maximum synergism in both strains. The results show that there is a correlation between the enzyme activity and tolerance, and carboxylic-esterase (Car-EST) is likely the main detoxification mechanism responsible for tolerance of S. littoralis to chlorantraniliprole.

Keywords: Chlorantraniliprole, detoxification enzymes, Egypt, Spodoptera littoralis.

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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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Authors: Paola Lecca, Lorenzo Dematte, Corrado Priami

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The present models and simulation algorithms of intracellular stochastic kinetics are usually based on the premise that diffusion is so fast that the concentrations of all the involved species are homogeneous in space. However, recents experimental measurements of intracellular diffusion constants indicate that the assumption of a homogeneous well-stirred cytosol is not necessarily valid even for small prokaryotic cells. In this work a mathematical treatment of diffusion that can be incorporated in a stochastic algorithm simulating the dynamics of a reaction-diffusion system is presented. The movement of a molecule A from a region i to a region j of the space is represented as a first order reaction Ai k- ! Aj , where the rate constant k depends on the diffusion coefficient. The diffusion coefficients are modeled as function of the local concentration of the solutes, their intrinsic viscosities, their frictional coefficients and the temperature of the system. The stochastic time evolution of the system is given by the occurrence of diffusion events and chemical reaction events. At each time step an event (reaction or diffusion) is selected from a probability distribution of waiting times determined by the intrinsic reaction kinetics and diffusion dynamics. To demonstrate the method the simulation results of the reaction-diffusion system of chaperoneassisted protein folding in cytoplasm are shown.

Keywords: Reaction-diffusion systems, diffusion coefficient, stochastic simulation algorithm.

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Authors: Nagasamy Venkatesh Dhandapani, Amged Awad El-Gied

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Cefixime, a BCS class II drug, is insoluble in water but freely soluble in acetone and in alcohol. The aqueous solubility of cefixime in water is poor and exhibits exceptionally slow and intrinsic dissolution rate. In the present study, cefixime and β-Cyclodextrin (β-CD) solid dispersions were prepared with a view to study the effect and influence of β-CD on the solubility and dissolution rate of this poorly aqueous soluble drug. Phase solubility profile revealed that the solubility of cefixime was increased in the presence of β-CD and was classified as A_L-type. Effect of variable, such as drug:carrier ratio, was studied. Physical characterization of the solid dispersion was characterized by Fourier transform infrared spectroscopy (FT-IR) and Differential scanning calorimetry (DSC). These studies revealed that a distinct loss of drug crystallinity in the solid molecular dispersions is ostensibly accounting for enhancement of dissolution rate in distilled water. The drug release from the prepared solid dispersion exhibited a first order kinetics. Solid dispersions of cefixime showed a 6.77 times fold increase in dissolution rate over the pure drug.

Keywords: Cefixime, β-Cyclodextrin, solid dispersions, kneading method, dissolution, release kinetics.

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Authors: L. N. Corîci, A. E. Frissen, D -J. Van Zoelen, I. F. Eggen, F. Peter, C. M. Davidescu, C. G. Boeriu

Abstract:

Two commercial proteases from Bacillus licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were screened for the production of Z-Ala-Phe-NH2 in batch reaction. Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal amidation of Z-Ala-Phe-OMe using ammonium carbamate as ammonium source. Immobilization of protease has been achieved by the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and tetramethoxysilane (TMOS) as precursors (unpublished results). In batch production, about 95% of Z-Ala-Phe-NH2 was obtained at 30°C after 24 hours of incubation. Reproducibility of different batches of commercial Alcalase 2.4 L FG preparations was also investigated by evaluating the amidation activity and the entrapment yields in the case of immobilization. A packed-bed reactor (0.68 cm ID, 15.0 cm long) was operated successfully for the continuous synthesis of peptide amides. The immobilized enzyme retained the initial activity over 10 cycles of repeated use in continuous reactor at ambient temperature. At 0.75 mL/min flow rate of the substrate mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5 hours of substrate recycling. The product contained about 90% peptide amide and 10% hydrolysis byproduct.

Keywords: packed-bed reactor, peptide amide, protease, sol-gel immobilization.

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Authors: Manjeet Bansal, Diwan Singh, V.K.Garg, Pawan Rose

Abstract:

The potential of economically cheaper cellulose containing natural materials like rice husk was assessed for nickel adsorption from aqueous solutions. The effects of pH, contact time, sorbent dose, initial metal ion concentration and temperature on the uptake of nickel were studied in batch process. The removal of nickel was dependent on the physico-chemical characteristics of the adsorbent, adsorbate concentration and other studied process parameters. The sorption data has been correlated with Langmuir, Freundlich and Dubinin-Radush kevich (D-R) adsorption models. It was found that Freundlich and Langmuir isotherms fitted well to the data. Maximum nickel removal was observed at pH 6.0. The efficiency of rice husk for nickel removal was 51.8% for dilute solutions at 20 g L-1 adsorbent dose. FTIR, SEM and EDAX were recorded before and after adsorption to explore the number and position of the functional groups available for nickel binding on to the studied adsorbent and changes in surface morphology and elemental constitution of the adsorbent. Pseudo-second order model explains the nickel kinetics more effectively. Reusability of the adsorbent was examined by desorption in which HCl eluted 78.93% nickel. The results revealed that nickel is considerably adsorbed on rice husk and it could be and economic method for the removal of nickel from aqueous solutions.

Keywords: Adsorption, nickel, SEM, EDAX.

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Authors: Asok Adak, Debabrata Mazumder, Pratip Bandyopadhyay

Abstract:

Anaerobic Digestion has become a promising technology for biological transformation of organic fraction of the municipal solid wastes (MSW). In order to represent the kinetic behavior of such biological process and thereby to design a reactor system, development of a mathematical model is essential. Addressing this issue, a simplistic mathematical model has been developed for anaerobic digestion of MSW in a continuous flow reactor unit under homogeneous steady state condition. Upon simulated hydrolysis, the kinetics of biomass growth and substrate utilization rate are assumed to follow first order reaction kinetics. Simulation of this model has been conducted by studying sensitivity of various process variables. The model was simulated using typical kinetic data of anaerobic digestion MSW and typical MSW characteristics of Kolkata. The hydraulic retention time (HRT) and solid retention time (SRT) time were mainly estimated by varying different model parameters like efficiency of reactor, influent substrate concentration and biomass concentration. Consequently, design table and charts have also been prepared for ready use in the actual plant operation.

Keywords: Anaerobic digestion, municipal solid waste (MSW), process design model, simulation study, hydraulic retention time(HRT), solid retention time (SRT).

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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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Authors: C. Zarcula, R. Croitoru, L. Corîci, C. Csunderlik, F. Peter

Abstract:

Lipases are enzymes particularly amenable for immobilization by entrapment methods, as they can work equally well in aqueous or non-conventional media and long-time stability of enzyme activity and enantioselectivity is needed to elaborate more efficient bioprocesses. The improvement of Pseudomonas fluorescens (Amano AK) lipase characteristics was investigated by optimizing the immobilization procedure in hybrid organic-inorganic matrices using ionic liquids as additives. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety are beneficial for the activity of immobilized lipase. Silanes with alkyl- or aryl nonhydrolizable groups used as precursors in combination with tetramethoxysilane could generate composites with higher enantioselectivity compared to the native enzyme in acylation reactions of secondary alcohols. The optimal effect on both activity and enantioselectivity was achieved for the composite made from octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60% increase of total activity following immobilization and enantiomeric ratio of 30). Ionic liquids also demonstrated valuable properties as reaction media for the studied reactions, comparable with the usual organic solvent, hexane.

Keywords: Ionic liquids, lipase, enantioselectivity, sol-gelimmobilization

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