Search results for: enzymatic treatment

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

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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: Payel Ghosh, Rama Chandra Pradhan, Sabyasachi Mishra

Abstract:

Jamun (Syzygium cuminii L.) is one of the important indigenous minor fruit with high medicinal value. The jamun cultivation is unorganized and there is huge loss of this fruit every year. The perishable nature of the fruit makes its postharvest management further difficult. Due to the strong cell wall structure of pectin-protein bonds and hard seeds, extraction of juice becomes difficult. Enzymatic treatment has been commercially used for improvement of juice quality with high yield. The objective of the study was to optimize the best treatment method for juice extraction. Enzymes (Pectinase and Tannase) from different stains had been used and for each enzyme, best result obtained by using response surface methodology. Optimization had been done on the basis of physicochemical property, nutritional property, sensory quality and cost estimation. According to quality aspect, cost analysis and sensory evaluation, the optimizing enzymatic treatment was obtained by Pectinase from Aspergillus aculeatus strain. The optimum condition for the treatment was 44 ^oC with 80 minute with a concentration of 0.05% (w/w). At these conditions, 75% of yield with turbidity of 32.21NTU, clarity of 74.39%T, polyphenol content of 115.31 mg GAE/g, protein content of 102.43 mg/g have been obtained with a significant difference in overall acceptability.

Keywords: Jamun, enzymatic treatment, physicochemical property, sensory analysis, optimization.

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Authors: Boonyisa Wanitwattanarumlug, Apanee Luengnaruemitchai, Sujitra Wongkasemjit

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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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Authors: S. Gaewchingduang, P. Pengthemkeerati

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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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Authors: Samera Salimpour Abkenar

Abstract:

In traditional dyeing of natural fibers with natural dyes, metal salts are commonly used to increase color stability. This method always carries the risk of environmental pollution (contamination of arable soils and fresh groundwater) due to the release of dyeing effluents containing large amounts of metal. Therefore, researchers are always looking for new methods to obtain a green dyeing system. In this research, the use of the enzymatic dyeing method to prevent environmental pollution with metals and reduce production costs has been proposed. After degumming and bleaching, raw silk fabrics were dyed with natural dyes (Madder and Sumac) by three methods (pre-mordanting with a metal salt, one-step enzymatic dyeing, and two-step enzymatic dyeing). Results show that silk dyed with natural dyes by the enzymatic method has higher color strength and colorfastness than the pretreated with a metal salt. Also, the amount of remained dyes in the dyeing wastewater is significantly reduced by the enzymatic method. It is found that the enzymatic dyeing method leads to improvement of dye absorption, color strength, soft hand, no change in color shade, low production costs (due to low dyeing temperature), and a significant reduction in environmental pollution.

Keywords: Eco-friendly, natural dyes, silk, traditional dyeing.

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Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej

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

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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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Authors: Kessara Seneesrisakul, Erdogan Gulari, Sumaeth Chavadej

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

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

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

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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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Authors: Khalida Boutemak, Nasssima Benali, Nadji Moulai-Mostefa

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Effect of enzyme on the yield and chemical composition of Artemisia campestris essential oil is reported in the present study. It was demonstrated that enzyme facilitated the extraction of essential oil with increase in oil yield and did not affect any noticeable change in flavour profile of the  volatile oil. Essential oil was tested for antibacterial activity using Escherichia coli; which was extremely sensitive against control with the largest inhibition (29mm), whereas Staphylococcus aureus was the most sensitive against essential oil obtained from enzymatic pre-treatment with the largest inhibition zone (25mm). The antioxidant activity of the essential oil with hemicellulase pre-treatment (EO2) and control sample (EO1) was determined through reducing power. It was significantly lower than the standard drug (vitamin C) in this order: vitamin C˃EO2˃EO1.

Keywords: Artemisia campestris, enzyme pre-treatment, hemicellulase, antibacterial activity, antioxidant activity.

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Authors: D.T. Mirzarakhmetova

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The studying of enzymatic esterification of carboxylic acids and higher alcohols was performed by esterase Saccharomyces cerevisiae in water-organic medium. Investigation of the enzyme specificity to acetic substrates showed the best result with acetic acid in esterification reactions with ethanol whereas within other carboxylic acids the esterification decreased with acids: hexanoic > pentanoic > butyric > decanoic. In relation to higher alcohols C3-C5, esterification increased with alcohols propanol < butanol < amylol. Also it was determined that esterase was more specific to alcohols with branched chain such as isobutyl alcohol and isoamyl alcohol. Data obtained may have important practical implications, for example, for application of yeast esterase in producing various volatile esters as well as in enzymatic transformation of volatile acids and toxic fusel alcohols into volatile esters by providing the production of the high quality alcoholic beverages with redused content of higher alcohols as well as with improved degustational and hygienic properties.

Keywords: enzymes in non-conventional media, esterification, higher alcohols, volatile esters, yeast esterase

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Authors: Shanmugasundaram Krishnakumari, Paramasivam Rajeswari, Subramanian Kathiravan

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Mushrooms are a group of fleshy macroscopic fungi. They have been valued throughout the world as both edible and medicine. They are highly nutritious with good amount of quality proteins, vitamins and minerals. An edible mushroom, Calocybe indica was selected to validate its nutritional and medicinal properties. Since tissue damage in hyperglycemia has been related to oxidative stress, we evaluated the enzymatic and non-enzymatic antioxidant status in the serum, liver and kidney since they are the target organs in diabetic complications. From the results, increased oxidative stress and decreased antioxidants might be related to the causation of diabetes mellitus. The treatment in the diabetic rats with the Calocybe indica showed an increase in the antioxidant system and decrease in the production of free radicals. The mushrooms which contain antioxidant phytochemicals has potential free radical scavenging capacity and hence can induce the antioxidant system in the body significantly reduces the generated free radicals thereby maintaining the normal levels of the antioxidants

Keywords: Antioxidants, Calocybe indica, diabetes mellitus, edible mushroom, oxidative stress.

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Authors: Ismail I. Abo Ghanema, Kadry M. Sadek

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Oxidative stress and overwhelming free radicals associated with diabetes mellitus are likely to be linked with development of certain complication such as retinopathy, nephropathy and neuropathy. Treatment of diabetic subjects with antioxidant may be of advantage in attenuating these complications. Olive leaf (Oleaeuropaea), has been endowed with many beneficial and health promoting properties mostly linked to its antioxidant activity. This study aimed to evaluate the significance of supplementation of Olive leaves extract (OLE) in reducing oxidative stress, hyperglycemia and hyperlipidemia in Sterptozotocin (STZ)- induced diabetic rats. After induction of diabetes, a significant rise in plasma glucose, lipid profiles except High density lipoproteincholestrol (HDLc), malondialdehyde (MDA) and significant decrease of plasma insulin, HDLc and Plasma reduced glutathione GSH as well as alteration in enzymatic antioxidants was observed in all diabetic animals. During treatment of diabetic rats with 0.5g/kg body weight of Olive leaves extract (OLE) the levels of plasma (MDA) ,(GSH), insulin, lipid profiles along with blood glucose and erythrocyte enzymatic antioxidant enzymes were significantly restored to establish values that were not different from normal control rats. Untreated diabetic rats on the other hand demonstrated persistent alterations in the oxidative stress marker (MDA), blood glucose, insulin, lipid profiles and the antioxidant parameters. These results demonstrate that OLE may be of advantage in inhibiting hyperglycemia, hyperlipidemia and oxidative stress induced by diabetes and suggest that administration of OLE may be helpful in the prevention or at least reduced of diabetic complications associated with oxidative stress.

Keywords: Diabetes mellitus, olive leaves, oxidative stress, red blood cells

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

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

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

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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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Authors: Vasudha Bansal, M. L. Singla, C. Ghanshyam

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The effect of nonthermal pulsed electric field (PEF) and thermal treatment (90⁰C for 60s) was studied on quality parameters of emblica officinalis juice for the period of 6 weeks at 4⁰C using monopolar rectangular pulse of 1µs width. The PEF treatment was given using static chamber at 24kV/cm for 500µs. The quality of emblica officinalis juice was investigated in terms of non enzymatic browning index (NEBI), 5-hydroxymethyl-2-furfural (HMF), total polyphenol content and antioxidant capacity. ⁰Brix, pH and conductivity were evaluated as physical parameters. The aim of the work was to investigate the effect of PEF on the retention of bioactive compounds and retardation of browning activity. The results showed that conventional thermal treatment had led to a significant (p < 0.05) decrease of 48.15% in polyphenol content (129.56 mg of GAE L^-1), with higher NEBI and HMF formation (p < 0.05) whilst PEF suppressed NEBI and retained higher polyphenol compounds (168.59 mg GAE L^-1) with limiting the loss to 32.56% along maximum free radical scavenging activity (92.07%). However, pH, ⁰brix and electrical conductivity of treated juice samples remain unaffected. Therefore, PEF can be considered as an effective nonthermal treatment for retaining bioactive compounds along suppressing browning of emblica juice.

 

Keywords: Emblica officinalis juice, Free radical scavenging activity, Pulsed electric field, Total polyphenol content.

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Authors: V. Radenkovs, D. Klava, K. Juhnevica

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The present study addresses problems and solutions related to new functional food production. Wheat (Triticum aestivum L) bran obtained from industrial mill company “Dobeles dzirnavieks”, was used to investigate them as raw material like nutrients for Bifidobacterium lactis Bb-12. Enzymatic hydrolysis of wheat bran starch was carried out by α-amylase from Bacillus amyloliquefaciens (Sigma Aldrich). The Viscozyme L purchased from (Sigma Aldrich) were used for reducing released sugar. Bifidibacterium lactis Bb-12 purchased from (Probio-Tec® CHR Hansen) was cultivated in enzymatically hydrolysed wheat bran mash. All procedures ensured the number of active Bifidobacterium lactis Bb-12 in the final product reached 105 CFUg-1. After enzymatic and bacterial fermentations sample were freeze dried for analysis of chemical compounds. All experiments were performed at Faculty of Food Technology of Latvia University of Agriculture in January- March 2013. The obtained results show that both types of wheat bran (enzymatically treated and non-treated) influenced the fermentative activity and number of Bifidibacterium lactis Bb-12 viable in wheat bran mash. Amount of acidity strongly increase during the wheat bran mash fermentation. The main objective of this work was to create low-energy functional enzymatically and bacterially treated food from wheat bran using enzymatic hydrolysis of carbohydrates and following cultivation of Bifidobacterium lactis Bb-12.

Keywords: Viscozyme L, α-amylase, Bifidobacterium lactis, fermented wheat bran.

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Authors: M. S. Kilic, S. Korkut, B. Hazer

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Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2 and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: Bilirubin oxidase, Enzymatic fuel cell, Glucose oxidase, Nanoparticles.

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Authors: R. Charoen, S. Tipkanon, W. Savedboworn, N. Phonsatta, A. Panya

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Rice bran protein hydrolysates (RBPH) were prepared from defatted rice bran of two different Thai rice cultivars (Plai-Ngahm-Prachinburi; PNP and Khao Dok Mali 105; KDM105) using an enzymatic method. This research aimed to optimize enzyme-assisted protein extraction. In addition, the functional properties of RBPH and their stabilities to environmental stresses including pH (3 to 8), ionic strength (0 mM to 500 mM) and the thermal treatment (30 °C to 90 °C) were investigated. Results showed that enzymatic process for protein extraction of defatted rice bran was as follows: enzyme concentration 0.075 g/ 5 g of protein, extraction temperature 50 °C and extraction time 4 h. The obtained protein hydrolysate powders had a degree of hydrolysis (%) of 21.05% in PNP and 19.92% in KDM105. The solubility of protein hydrolysates at pH 4-6 was ranged from 27.28-38.57% and 27.60-43.00% in PNP and KDM105, respectively. In general, antioxidant activities indicated by total phenolic content, FRAP, ferrous ion-chelating (FIC), and 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) of KDM105 had higher than PNP. In terms of functional properties, the emulsifying activity index (EAI) was was 8.78 m²/g protein in KDM105, whereas PNP was 5.05 m²/g protein. The foaming capacity at 5 minutes (%) was 47.33 and 52.98 in PNP and KDM105, respectively. Glutamine, Alanine, Valine, and Leucine are the major amino acid in protein hydrolysates where the total amino acid of KDM105 gave higher than PNP. Furthermore, we investigated environmental stresses on the stability of 5% oil in water emulsion (5% oil, 10 mM citrate buffer) stabilized by RBPH (3.5%). The droplet diameter of emulsion stabilized by KDM105 was smaller (d < 250 nm) than produced by PNP. For environmental stresses, RBPH stabilized emulsions were stable at pH around 3 and 5-6, at high salt (< 400 mM, pH 7) and at temperatures range between 30-50°C.

Keywords: Functional properties, oil in water emulsion, protein hydrolysates, rice bran protein.

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Authors: Anna Trusek-Holownia, Andrzej Noworyta

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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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Authors: Pilanee Vaithanomsat, Sinsupha Chuichulcherm, Waraporn Apiwatanapiwat

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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: Y.-H Tshela Ntumba, A. Przepiórkowska, M. Prochoń

Abstract:

Polymeric materials have become an integral part of every aspect of today's industry. They have wide applications, inter alia, in areas such as medicine, food industry and agriculture. In agriculture, for example, they are used for the production of pots, irrigation systems and for soil mulching. The aim of this study was the attempt to produce a biodecomposable agricultural mat, by coating cotton fabric with a blend of carboxylated styrene-butadiene latex (LBSK) containing the enzymatic hydrolyzate of keratin from cattle hair, which would serve as a material for mulching.

The production of such material allows the beneficial management of burdensome tannery waste constituted by keratin from cattle hair and at the same time, the production of agricultural mats that much faster undergo decomposition than commonly used polyethylene mats.

Keywords: Agricultural mat, biodecomposition, biodegradation, carboxylated styrene-butadiene latex, cellulosic-elastomeric material, keratin hydrolyzate, mulching, protein hydrolyzate.

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Authors: Jirakarn Nantapipat, Apanee Luengnaruemitchai, Sujitra Wongkasemjit

Abstract:

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

Keywords: Corncobs, Pretreatment, Sulfuric acid, Phosphoric acid.

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Authors: Sirikarn Satimanont, Apanee Luengnaruemitchai, Sujitra Wongkasemjit

Abstract:

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

Keywords: Butanol production, Corn cobs, Phosphoric acid, Pretreatment

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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: 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: M. Caramihai, Irina Severin, H. Balan, A. Blidaru, V. Balanica

Abstract:

Accurate assessment of the primary tumor response to treatment is important in the management of breast cancer. This paper introduces a new set of treatment evaluation indicators for breast cancer cases based on the computational process of three known metrics, the Euclidian, Hamming and Levenshtein distances. The distance principals are applied to pairs of mammograms and/or echograms, recorded before and after treatment, determining a reference point in judging the evolution amount of the studied carcinoma. The obtained numerical results are indeed very transparent and indicate not only the evolution or the involution of the tumor under treatment, but also a quantitative measurement of the benefit in using the selected method of treatment.

Keywords: Breast cancer, Distance metrics, Cancer treatment evaluation.

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