Search results for: enzymatic treatment

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: Sasiporn Rattanasupha, Settapat Chinviriyasit

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The treatment function adopts a continuous and differentiable function which can describe the effect of delayed treatment when the number of infected individuals increases and the medical condition is limited. In this paper, the SEIR epidemic model with treatment function is studied to investigate the dynamics of the model due to the effect of treatment. It is assumed that the treatment rate is proportional to the number of infective patients. The stability of the model is analyzed. The model is simulated to illustrate the analytical results and to investigate the effects of treatment on the spread of infection.

Keywords: basic reproduction number, local stability, SEIR epidemic model, treatment function

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Authors: Meha Alouini, Wissem Mnif, Yasmine Souissi

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This work will be conducted within the framework of the environmental sustainable development. It involves waste recovering into biodiesel fuel. Low cost feedstocks such as waste of frying oil and animal fats have been utilized to replace refined vegetable oil for biodiesel production. Biodiesel which refers to fatty acid methyl esters (FAME) was carried out by both chemical and enzymatic reaction of transesterification. In order to compare the two studied reactions the obtained biodiesel was characterized by determining its esters content and its fuel properties according to the European standard EN 14214. It was noted that the chemical method gave the product with the best physical property. But the biological one was found more effective for obtaining important ester content. Thus it would be interesting to optimize the enzymatic pathway of production of biodiesel to obtain a better property of biodiesel.

Keywords: biodiesel, fatty acid methyl esters, transesterification, waste frying oil, waste beef fat

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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: Mustafa Ümi̇t Ünal, Nafi̇z Çeli̇ktaş, Aysun Şener, Sara Betül Dolgun, Duygu Keser

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The aim of this study was to compare acid and base pretreatment of switchgrass for bioethanol production. Switchgrass was pretreated with sulfuric acid and sodium hydroxide at 0.5, 1.0 and 1.5% (v/v) at 120, 140, 180 °C for 10, 60 and 90. Optimization of enzymatic hydrolysis of the pretreated switchgrass samples were carried out using three different enzyme mixtures (22.5 mg cellulase and 75 mg cellobiase /g biomass; 45 mg cellulase and 150 mg cellobiase /g biomass; 90 mg cellulase and 300 mg cellobiase /g biomass). Samples were removed at 24-h interval for fermentable sugar analyses with HPLC. The results showed that use of 90 mg cellulase and 300 mg cellobiase/g biomass resulted in the highest fermentable sugar formation. Furthermore, the highest fermentable sugar yield was obtained by pretreatment at 120 °C for 10 min using 1.0 % sodium hydroxide.

Keywords: switchgrass, acid pretreatment, enzymatic hydrolysis, base pretreatment, ethanol production

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

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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 butadiene-styrene latex, cellulosic-elastomeric material, keratin hydrolyzate, mulching, protein hydrolyzate

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Authors: Ajay Kumar Singh, Arvind Kumar, S. P. Singh

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Cellulose nanofibrils (CNF) have shown potential of their extensive use in various fields, including papermaking, due to their unique characteristics. In this study, CNF’s were prepared by fibrillating the pulp obtained from raw materials e.g. bagasse, hardwood and softwood using enzymatic pretreatment followed by mechanical refining. These nanofibrils, when examined under FE-SEM, show that partial fibrillation on fiber surface has resulted in production of nanofibers. Mixing these nanofibers with the unrefined and normally refined fibers show their reinforcing effect. This effect is manifested in observing the improvement in the physical and mechanical properties e.g. tensile index and burst index of paper. Tear index, however, was observed to decrease on blending with nanofibers. The optical properties of paper sheets made from blended fibers showed no significant change in comparison to those made from only mechanically refined pulp. Mixing of normal pulp fibers with nanofibers show increase in ºSR and consequent decrease in drainage rate. These changes observed in mechanical, optical and other physical properties of the paper sheets made from nanofibrils blended pulp have been tried to explain considering the distribution of the nanofibrils alongside microfibrils in the fibrous network. Since usually, paper/boards with higher strength are observed to have diminished optical properties which is a drawback in their quality, the present work has the potential for developing paper/boards having improved strength alongwith undiminished optical properties utilising the concepts of nanoscience and nanotechnology.

Keywords: enzymatic pretreatment, mechanical refining, nanofibrils, paper properties

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Authors: Keyur Raval, Steffen Krohn, Bruno Moerschbacher

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Chitin, the biopolymer of the N-acetylglucosamine, is the most abundant biopolymer on the planet after cellulose. Industrially, chitin is isolated and purified from the shell residues of shrimps. A deacetylated derivative of chitin i.e. chitosan has more market value and applications owing to it solubility and overall cationic charge compared to the parent polymer. This deacetylation on an industrial scale is performed chemically using alkalis like sodium hydroxide. This reaction not only is hazardous to the environment owing to negative impact on the marine ecosystem. A greener option to this process is the enzymatic process. In nature, the naïve chitin is converted to chitosan by chitin deacetylase (CDA). This enzymatic conversion on the industrial scale is however hampered by the crystallinity of chitin. Thus, this enzymatic action requires the substrate i.e. chitin to be soluble which is technically difficult and an energy consuming process. We in this project wanted to address this shortcoming of CDA. In lieu of this, we have modeled a fusion protein with CDA and an auxiliary protein. The main interest being to increase the accessibility of the enzyme towards crystalline chitin. A similar fusion work with chitinases had improved the catalytic ability towards insoluble chitin. In the first step, suitable partners were searched through the protein data bank (PDB) wherein the domain architecture were sought. The next step was to create the models of the fused product using various in silico techniques. The models were created by MODELLER and evaluated for properties such as the energy or the impairment of the binding sites. A fusion PCR has been designed based on the linker sequences generated by MODELLER and would be tested for its activity towards insoluble chitin.

Keywords: chitin deacetylase, modeling, chitin binding domain, chitinases

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Authors: Lucretia Ramnath

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Fishmeal is extensively used for fish farming but is an expensive fish feed ingredient. A cheaper alternate to fishmeal is single cell protein (SCP) which can be cultivated on fermentable sugars recovered from organic waste streams such as pulp and paper mill sludge (PPMS). PPMS has a high cellulose content, thus is suitable for glucose recovery through enzymatic hydrolysis but is hampered by lignin and ash. To render PPMS amenable for enzymatic hydrolysis, the PPMS waspre-treated to produce a glucose-rich hydrolysate which served as a feed stock for the production of fungal SCP. The PPMS used in this study had the following composition: 72.77% carbohydrates, 8.6% lignin, and 18.63% ash. The pre-treatments had no significant effect on lignin composition but had a substantial effect on carbohydrate and ash content. Enzymatic hydrolysis of screened PPMS was previously optimized through response surface methodology (RSM) and 2-factorial design. The optimized protocol resulted in a hydrolysate containing 46.1 g/L of glucose, of which 86% was recovered after downstream processing by passing through a 100-mesh sieve (38 µm pore size). Vogel’s medium supplemented with 10 g/L hydrolysate successfully supported the growth of Fusarium venenatum, conducted using standard growth conditions; pH 6, 200 rpm, 2.88 g/L ammonium phosphate, 25°C. A maximum F. venenatum biomass of 45 g/L was produced with a yield coefficient of 4.67. Pulp and paper mill sludge hydrolysate contained approximately five times more glucose than what was needed for SCP production and served as a suitable carbon source. We have shown that PPMS can be successfully beneficiated for SCP production.

Keywords: pulp and paper waste, fungi, single cell protein, hydrolysate

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Authors: Preshanthan Moodley, E. B. Gueguim Kana

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This study examines the kinetics of S. cerevisiae BY4743 growth and bioethanol production from sugarcane leaf waste (SLW), utilizing two different optimized pretreatment regimes; under two fermentation modes: steam salt-alkali filtered enzymatic hydrolysate (SSA-F), steam salt-alkali unfiltered (SSA-U), microwave salt-alkali filtered (MSA-F) and microwave salt-alkali unfiltered (MSA-U). The kinetic coefficients were determined by fitting the Monod, modified Gompertz, and logistic models to the experimental data with high coefficients of determination R² > 0.97. A maximum specific growth rate (µₘₐₓ) of 0.153 h⁻¹ was obtained under SSA-F and SSA-U whereas, 0.150 h⁻¹ was observed with MSA-F and MSA-U. SSA-U gave a potential maximum bioethanol concentration (Pₘ) of 31.06 g/L compared to 30.49, 23.26 and 21.79g/L for SSA-F, MSA-F and MSA-U respectively. An insignificant difference was observed in the μmax and Pm for the filtered and unfiltered enzymatic hydrolysate for both SSA and MSA pretreatments, thus potentially reducing a unit operation. These findings provide significant insights for process scale up.

Keywords: lignocellulosic bioethanol, microwave pretreatment, sugarcane leaves, kinetics

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Authors: A. Zh. Aupova, A. Ulankyzy, A. Sarsenova, A. Kussayin, Sh. Turarbek, N. Moldagulova, A. Kurmanbayev

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One of the methods of organic waste bioconversion into environmentally-friendly fertilizer is composting. Microorganisms that produce hydrolytic enzymes play a significant role in accelerating the process of organic waste composting. We studied the enzymatic potential (amylase, protease, cellulase, lipase, urease activity) of bacteria isolated from the sewage sludge of Nur-Sultan, Rudny, and Fort-Shevchenko cities, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha for processing organic waste and identifying active strains. Microorganism isolation was carried out by the cultures enrichment method on liquid nutrient media, followed by inoculating on different solid media to isolate individual colonies. As a result, sixty-one microorganisms were isolated, three of which were thermophiles (DS1, DS2, and DS3). The highest number of isolates, twenty-one and eighteen, were isolated from sewage sludge of Nur-Sultan and Rudny cities, respectively. Ten isolates were isolated from the wastewater of the sewage treatment plant in Fort-Shevchenko. From the dacha soil of Nur-Sultan city and freshly cut grass - 9 and 5 isolates were revealed, respectively. The lipolytic, proteolytic, amylolytic, cellulolytic, ureolytic, and oil-oxidizing activities of isolates were studied. According to the results of experiments, starch hydrolysis (amylolytic activity) was found in 2 isolates - CB2/2, and CB2/1. Three isolates - CB2, CB2/1, and CB1/1 were selected for the highest ability to break down casein. Among isolated 61 bacterial cultures, three isolates could break down fats - CB3, CBG1/1, and IL3. Seven strains had cellulolytic activity - DS1, DS2, IL3, IL5, P2, P5, and P3. Six isolates rapidly decomposed urea. Isolate P1 could break down casein and cellulose. Isolate DS3 was a thermophile and had cellulolytic activity. Thus, based on the conducted studies, 15 isolates were selected as a potential for sewage sludge composting - CB2, CB3, CB1/1, CB2/2, CBG1/1, CB2/1, DS1, DS2, DS3, IL3, IL5, P1, P2, P5, P3. Selected strains were identified on a mass spectrometer (Maldi-TOF). The isolate - CB 3 was referred to the genus Rhodococcus rhodochrous; two isolates CB2 and CB1 / 1 - to Bacillus cereus, CB 2/2 - to Cryseobacterium arachidis, CBG 1/1 - to Pseudoxanthomonas sp., CB2/1 - to Bacillus megaterium, DS1 - to Pediococcus acidilactici, DS2 - to Paenibacillus residui, DS3 - to Brevibacillus invocatus, three strains IL3, P5, P3 - to Enterobacter cloacae, two strains IL5, P2 - to Ochrobactrum intermedium, and P1 - Bacillus lichenoformis. Hence, 60 isolates were isolated from the wastewater of the cities of Nur-Sultan, Rudny, Fort-Shevchenko, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha. Based on the highest enzymatic activity, 15 active isolates were selected and identified. These strains may become the candidates for bio preparation for sewage sludge processing.

Keywords: sewage sludge, composting, bacteria, enzymatic activity

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Authors: H. Djebar, L. Khene, M. Boucenna, M. R. Djebar, M. N. Khebbeb, M. Djekoun

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Currently in toxicology, use of alternative models permits to understand the mechanisms of toxicity at different levels of cells. Objectives of our research concern the determination of NPs ZnO, TiO2, AlO2, and FeO2 effect on ciliate protist freshwater Paramecium sp and Helix aspersa. The result obtained show that NPs increased antioxidative enzyme activity like catalase, glutathione –S-transferase and level GSH. Also, cells treated with high concentrations of NPs showed a high level of MDA. In conclusion, observations from growth and enzymatic parameters suggest on one hand that treatment with NPs provokes an oxidative stress and on the other that snale and paramecium are excellent alternatives models for ecotoxicological studies.

Keywords: NPs, GST, catalase, GSH, MDA, toxicity, snale and paramecium

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Authors: Harold Jideofor

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Wastewater treatment consists of applying known technology to improve or upgrade the quality of a wastewater. Usually wastewater treatment will involve collecting the wastewater in a central, segregated location (the Wastewater Treatment Plant) and subjecting the wastewater to various treatment processes. Most often, since large volumes of wastewater are involved, treatment processes are carried out on continuously flowing wastewaters (continuous flow or "open" systems) rather than as "batch" or a series of periodic treatment processes in which treatment is carried out on parcels or "batches" of wastewaters. While most wastewater treatment processes are continuous flow, certain operations, such as vacuum filtration, involving storage of sludge, the addition of chemicals, filtration and removal or disposal of the treated sludge, are routinely handled as periodic batch operations.

Keywords: wastewater treatment, environmental engineering, waste water

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Authors: Nurul Aqilah Mohd Zaini, Afroditi Chatzifragkou, Dimitris Charalampopoulos

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D-Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, Polylactic Acid (PLA). In this study, D-lactic acid was produced in microbial cultures using Lactobacillus coryniformis subsp. torquens as D-lactic acid producer and hydrolysates of Dried Distillers Grains with Solubles (DDGS) as fermentation substrate. Prior to fermentation, DDGS was first alkaline pretreated with 5% (w/v) NaOH, for 15 minutes (121oC/ ~16 psi). This led to the generation of DDGS solid residues, rich in carbohydrates and especially cellulose (~52%). The carbohydrate-rich solids were then subjected to enzymatic hydrolysis with Accellerase® 1500. For Separate Hydrolysis and Fermentation (SHF), enzymatic hydrolysis was carried out at 50oC for 24 hours, followed by fermentation of D-lactic acid at 37oC in controlled pH 6. The obtained hydrolysate contained 24 g/l glucose, 5.4 g/l xylose and 0.6 g/l arabinose. In the case of Simultaneous Saccharification and Fermentation (SSF), hydrolysis and fermentation were conducted in a single step process at 37oC in pH 5. The enzymatic hydrolysis of DGGS pretreated solids took place mostly during lag phase of L. coryniformis fermentation, with only a small amount of glucose consumed during the first 6 h. When exponential phase was started, glucose generation reduced as the microorganism started to consume glucose for D-lactic acid production. Higher concentrations of D-lactic acid were produced when SSF approach was applied, with 28 g/l D-lactic acid after 24 h of fermentation (84.5% yield). In contrast, 21.2 g/l D-lactic acid were produced when SHF was used. The optical pu rity of D-lactic acid produced from both experiments was 99.9%. Besides, approximately 2 g/l acetic acid was also generated due to lactic acid degradation after glucose depletion in SHF. SSF was proved an efficient towards DDGS ulilisation and D-lactic acid production, by reducing the overall processing time, yielding sufficient D-lactic acid concentrations without the generation of fermentation by-products.

Keywords: DDGS, alkaline pretreatment, SSF, D-lactic acid

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Authors: Abdulmannan Fadel

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Arabinoxylan is a non-starch polysaccharide (NSP), which is one of the most important polysaccharides contained within cereal grains. Wheat endosperm pentosan and rice bran contain a significant amount of arabinoxylan (7% in rice bran and 10-12% in wheat endosperm pentosan). Several methods have been used for arabinoxylan extraction with varying degrees of success e.g. enzymatic and alkaline treatment. Yet, the use of extrusion alone as a pre-treatment to increase the yield and reduce the molecular weight in wheat endosperm pentosan and rice bran has not been investigated. The samples (wheat pentosan and rice bran) were extruded using a Twin-screw extruder at a range of screw speeds (80 and 160 rpm) and barrel temperatures range (80 to 140°C) with a throughput of 30 Kg hr-1 and moisture content of 25%. Arabinoxylans were extracted with water and the extraction yield and molecular weight was determined using size exclusion high-pressure liquid chromatography system. It was found that increasing screw speed from 80 rpm to 160 rpm, did not effect the extraction yield (p < 0.05) of arabinoxylan from either the wheat endosperm pentosan or the rice bran. However, the molecular weight of the extracted arabinoxylans from pentosan was found to decrease with increasing screw speed in wheat endosperm pentosan. These low molecular weight arabinoxylans have been suggested as immunomodulators.

Keywords: arabinoxylans, extrusion, wheat endosperm pentosan, rice bran

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Authors: A. T. Ramachandra Naik, H. Shivananda Murthy, H. n. Anjanayappa

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The freshwater prawn, Macrobrachium rosenbergii is a more popular crustacean cultured widely in monoculture system in India. It has got high nutritional value in the human diet. Hence, understanding its enzymatic and body composition is important in order to judge its flesh quality. Fish oil specially derived from Indian oil sardine is a good source of highly unsaturated fatty acid and lipid source in fish/prawn diet. A 35% crude protein diet with graded levels of Sardine oil as a source of fat was incorporated at four levels viz, 2.07, 4.07, 6.07 and 8.07% maintaining a total lipid level of feed at 8.11, 10.24, 12.28 and 14.33% respectively. Diet without sardine oil (6.05% total lipid) was served as basal treatment. The giant freshwater prawn, Macrobrachium rosenbergii was used as test animal and the experiment was lost for 112 days. Significantly, higher gain in weight of prawn was recorded in the treatment with 6.07% sardine oil incorporation followed by higher specific growth rate, food conversion rate and protein efficiency ratio. The 8.07% sardine oil diet produced the highest RNA: DNA ratio in the prawn muscle. Digestive enzyme analyses in the digestive tract and mid-gut gland showed the greatest activity in prawns fed the 8.07% diet.

Keywords: digestive enzyme, fish diet, Macrobrachium rosenbergii, sardine oil

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Authors: M. Farhaoui, M. Derraz

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In the drinking water treatment processes, the optimization of the treatment is an issue of particular concern. In general, the process consists of many units as settling, coagulation, flocculation, sedimentation, filtration and disinfection. The optimization of the process consists of some measures to decrease the managing and monitoring expenses and improve the quality of the produced water. The objective of this study is to provide water treatment operators with methods and practices that enable to attain the most effective use of the facility and, in consequence, optimize the of the cubic meter price of the treated water. This paper proposes a review on optimization of drinking water treatment process by analyzing all of the water treatment units and gives some solutions in order to maximize the water treatment performances without compromising the water quality standards. Some solutions and methods are performed in the water treatment plant located in the middle of Morocco (Meknes).

Keywords: coagulation process, optimization, turbidity removal, water treatment

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

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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: Bengi Hakguder Taze, Sevcan Unluturk

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Şalak apricot (Prunus armeniaca L., cv. Şalak) is a specific variety grown in Igdir, Turkey. The fruit has distinctive properties distinguish it from other cultivars, such as its unique size, color, taste and higher water content. Drying is the widely used method for preservation of apricots. However, fresh consumption is preferred for Şalak apricot instead of drying due to its low dry matter content. Higher amounts of water in the structure and climacteric nature make the fruit sensitive against rapid quality loss during storage. Hence, alternative processing methods need to be introduced to extend the shelf life of the fresh produce. Mild heat (MH) treatment is of great interest as it can reduce the microbial load and inhibit enzymatic activities. Therefore, the aim of this study was to evaluate the impact of mild heat treatment on the natural microflora found on Şalak apricot surfaces and some physical quality parameters of the fruit, such as color and firmness. For this purpose, apricot samples were treated at different temperatures between 40 and 60 ℃ for different periods ranging between 10 to 60 min using a temperature controlled water bath. Natural flora on the fruit surfaces was examined using standard plating technique both before and after the treatment. Moreover, any changes in color and firmness of the fruit samples were also monitored. It was found that control samples were initially containing 7.5 ± 0.32 log CFU/g of total aerobic plate count (TAPC), 5.8±0.31 log CFU/g of yeast and mold count (YMC), and 5.17 ± 0.22 log CFU/g of coliforms. The highest log reductions in TAPC and YMC were observed as 3.87-log and 5.8-log after the treatments at 60 ℃ and 50 ℃, respectively. Nevertheless, the fruit lost its characteristic aroma at temperatures above 50 ℃. Furthermore, great color changes (ΔE ˃ 6) were observed and firmness of the apricot samples was reduced at these conditions. On the other hand, MH treatment at 41 ℃ for 10 min resulted in 1.6-log and 0.91-log reductions in TAPC and YMC, respectively, with slightly noticeable changes in color (ΔE ˂ 3). In conclusion, application of temperatures higher than 50 ℃ caused undesirable changes in physical quality of Şalak apricots. Although higher microbial reductions were achieved at those temperatures, temperatures between 40 and 50°C should be further investigated considering the fruit quality parameters. Another strategy may be the use of high temperatures for short time periods not exceeding 1-5 min. Besides all, MH treatment with UV-C light irradiation can be also considered as a hurdle strategy for better inactivation results.

Keywords: color, firmness, mild heat, natural flora, physical quality, şalak apricot

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Authors: Zintle Kolo, Ndiko Ludidi

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The enzymatic activity of p-coumarate 3-hydroxylase (C3H) synthesize caffeic acid from p-coumaric acid. We recently showed that exogenously applied caffeic acid confers salinity tolerance in soybean (Glycine max) by inducing antioxidant enzymatic activity to promote enhanced scavenging or reactive oxygen species, thus limiting salinity-induced oxidative stress. Recent evidence also establishes that pre-treatment of plants with exogenously supplied caffeic acid improves plant tolerance to osmotic stress by improving plant antioxidant capacity and enhancing biosynthesis of compatible solutes. We aimed to identify a C3H in maize (Zea mays) and evaluate the effect of drought on the spatial and temporal expression of the gene encoding the candidate maize C3H (ZmC3H). Primary sequence analysis shows that ZmC3H shares 71% identity with an Arabidopsis thaliana C3H that is implicated in the control of Arabidopsis cell expansion, growth, and responses to stress. In silico ZmC3H promoter analysis reveals the presence of cis-acting elements that interact with transcription factors implicated in plant responses to drought. Spatial expression analysis by semi-quantitative RT-PCR shows that ZmC3H is expressed in both leaves and roots under normal conditions. However, drought represses the expression of ZmC3H in leaves whereas it up-regulates its expression in roots. These changes in ZmC3H expression correlate with the changes in the content of caffeic acid in maize in response to drought. We illustrate the implications of these changes in the expression of the gene in relation to maize responses to drought and discuss the potential of regulating caffeic acid biosynthesis towards genetic improvement of maize tolerance to drought stress. These findings have implications for food security because of the potential of the implications of the study for drought tolerance in maize.

Keywords: caffeic acid, drought-responsive expression, maize drought tolerance, p-coumarate 3-hydroxylase

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

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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: Tatiana Franco, Hugo A. Estupinan

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Banana pseudostem fiber (BPF) and pineapple leaf fiber (PLF) for their excellent mechanical properties and biodegradability characteristics arouse interest in different areas of research. F In tropical regions, where the banana pseudostem and the pineapple leaf are transformed into hard-to-handle solid waste, they can be low-cost raw material and environmentally sustainable in research for composite materials. In terms of functionality of this type of fiber, an open structure would allow the adsorption and retention of organic, inorganic and metallic species. In general, natural fibers have closed structures on their surface with intricate internal arrangements that can be used for the solution of environmental problems and other technological uses, however it is not possible to access their internal structure and sublayers, exposing the fibers in the natural state. An alternative method to chemical and enzymatic treatment are the processes with the plasma treatments, which are known to be clean, economical and controlled. In this type of treatment, a gas contained in a reactor in the form of plasma acts on the fiber generating changes in its structure, morphology and topography. This work compares the effects on fibers of PLF and BPF treated with cold argon plasma, alternating time and current. These fibers are grown in the regions of Antioquia-Colombia. The morphological, compositional and wettability properties of the fibers were analyzed by Raman microscopy, contact angle measurements, scanning electron microscopy (SEM) and atomic force microscopy analysis (AFM). The treatment with cold plasma on PLF and BPF allowed increasing its wettability, the topography and the microstructural relationship between lignin and cellulose.

Keywords: cold plasma, contact angle, natural fibers, Raman, SEM, wettability

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Authors: Ouchemoukh Salim, Amessis-Ouchemoukh Nadia, Guenaoui Nawel, Moumeni Lynda, Zaidi Hicham, Otmani Amar, Sadou Dyhia

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Honey is a hive food rich in carbohydrates and water and it also has a lot of nutrients (enzymes, minerals, organic acids, phytochemicals...). It is used in different nutritional and therapeutic fields. Algerian honey was studied for its physicochemical parameters, nutritional values (moisture, brix, pH, electrical conductivity, and amounts of HMF, proteins, proline, total phenolic compounds and flavonoids) and some biological activities (antioxidant, anti-inflammatory and enzymatic anti-browning). The antioxidant activities of the samples were estimated using different methods (ABTS, DPPH free radicals scavenging, reducing power, and chelating ferrous activity). All honeys were acidic (3.45≤pH≤4.65). The color varied from mimosa yellow to dark brown. The specific rotation was levorotatory in most honey samples, and the electrical conductivity, hydroxymethylfurfural, and proline values agreed with the international honey requirements. For anti-inflammatory activity, the results showed that the inhibiting capacity of the denaturation of the BSA of the honey analyzed varied from 15 to 75 % with a maximum of activity at the concentration of 0,5 mg/ml. All honey exhibited enzymatic anti-browning on different slices of fruits. In fact, the results showed that the controls have the greatest browning unit compared to the honeys studied and PPO and POD enzymes had the lowest enzyme activity. High significant correlations were found between the color of honey, its antioxidant content and its biological activities (antioxidant, anti-inflammatory and enzymatic anti-browning). The dark color of honey is a good indicator of the best biological properties, therefore, the best nutritional and therapeutic values.

Keywords: honey, physico-chemical parameters, bioactive compounds, biological properties

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Authors: Shady S. Hassan, Brijesh K. Tiwari, Gwilym A. Williams, Amit K. Jaiswal

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Waste materials from a wide range of agro-industrial processes may be used as substrates for microbial growth, and subsequently the production of a range of high value products and bioenergy. In addition, utilization of these agro-residues in bioprocesses has the dual advantage of providing alternative substrates, as well as solving their disposal problems. Spent coffee grounds (SCG) are a by-product (45%) of coffee processing. SCG is a lignocellulosic material, which is composed mainly of cellulose, hemicelluloses, and lignin. Thus, a pretreatment process is required to facilitate an efficient enzymatic hydrolysis of such carbohydrates. In this context, microwave pretreatment of lignocellulosic biomass without the addition of harsh chemicals represents a green technology. Moreover, microwave treatment has a high heating efficiency and is easy to implement. Thus, microwave pretreatment of SCG without adding of harsh chemicals investigated as a green technology to enhance enzyme hydrolysis. In the present work, microwave pretreatment experiments were conducted on SCG at varying power levels (100, 250, 440, 600, and 1000 W) for 60 s. By increasing microwave power to a certain level (which vary by varying biomass), reducing sugar increases, then reducing sugar from biomass start to decrease with microwave power increase beyond this level. Microwave pretreatment of SCG at 60s followed by enzymatic hydrolysis resulted in total reducing sugars of 91.6 ± 7.0 mg/g of biomass (at microwave power of 100 w). Fourier transform Infrared Spectroscopy (FTIR) was employed to investigate changes in functional groups of biomass after pretreatment, while high-performance liquid chromatography (HPLC) was employed for determination of glucose. Pretreatment of lignocellulose using microwave was found to be an effective and energy efficient technology to improve saccharification and glucose yield. Energy performance will be evaluated for the microwave pretreatment, and the enzyme hydrolysate will be used as media component substitute for the production of ethanol and other high value products.

Keywords: lignocellulose, microwave, pretreatment, spent coffee grounds

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

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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

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Authors: Sanaz Alizadeh, Yves Comeau, Arshath Abdul Rahim, Sunhasis Ghoshal

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The application of silver nanoparticles (AgNPs) in personal care products, various household and industrial products has resulted in an inevitable environmental exposure of such engineered nanoparticles (ENPs). Ag ENPs, released via household and industrial wastes, reach water resource recovery facilities (WRRFs), yet the fate and transport of ENPs in WRRFs and their potential risk in the biological wastewater processes are poorly understood. Accordingly, our main objective was to elucidate the impact of long-term continuous exposure to AgNPs on biological activity of aerobic wastewater biofilm. The fate, transport and toxicity of 10 μg.L-1and 100 μg.L-1 PVP-stabilized AgNPs (50 nm) were evaluated in an attached growth biological treatment process, using lab-scale moving bed bioreactors (MBBRs). Two MBBR systems for organic matter removal were fed with a synthetic influent and operated at a hydraulic retention time (HRT) of 180 min and 60% volumetric filling ratio of Anox-K5 carriers with specific surface area of 800 m2/m3. Both reactors were operated for 85 days after reaching steady state conditions to develop a mature biofilm. The impact of AgNPs on the biological performance of the MBBRs was characterized over a period of 64 days in terms of the filtered biodegradable COD (SCOD) removal efficiency, the biofilm viability and key enzymatic activities (α-glucosidase and protease). The AgNPs were quantitatively characterized using single-particle inductively coupled plasma mass spectroscopy (spICP-MS), determining simultaneously the particle size distribution, particle concentration and dissolved silver content in influent, bioreactor and effluent samples. The generation of reactive oxygen species and the oxidative stress were assessed as the proposed toxicity mechanism of AgNPs. Results indicated that a low concentration of AgNPs (10 μg.L-1) did not significantly affect the SCOD removal efficiency whereas a significant reduction in treatment efficiency (37%) was observed at 100 μg.L-1AgNPs. Neither the viability nor the enzymatic activities of biofilm were affected at 10 μg.L-1AgNPs but a higher concentration of AgNPs induced cell membrane integrity damage resulting in 31% loss of viability and reduced α-glucosidase and protease enzymatic activities by 31% and 29%, respectively, over the 64-day exposure period. The elevated intercellular ROS in biofilm at a higher AgNPs concentration over time was consistent with a reduced biological biofilm performance, confirming the occurrence of a nanoparticle-induced oxidative stress in the heterotrophic biofilm. The spICP-MS analysis demonstrated a decrease in the nanoparticles concentration over the first 25 days, indicating a significant partitioning of AgNPs into the biofilm matrix in both reactors. The concentration of nanoparticles increased in effluent of both reactors after 25 days, however, indicating a decreased retention capacity of AgNPs in biofilm. The observed significant detachment of biofilm also contributed to a higher release of nanoparticles due to cell-wall destabilizing properties of AgNPs as an antimicrobial agent. The removal efficiency of PVP-AgNPs and the biofilm biological responses were a function of nanoparticle concentration and exposure time. This study contributes to a better understanding of the fate and behavior of AgNPs in biological wastewater processes, providing key information that can be used to predict the environmental risks of ENPs in aquatic ecosystems.

Keywords: biofilm, silver nanoparticle, single particle ICP-MS, toxicity, wastewater

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Authors: Joseph A. Bentil, Anders Thygesen, Lene Langea, Moses Mensah, Anne Meyer

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The study investigated the saccharification potential of in-house enzymes produced from a white-rot basidiomycete strain, Polyporus ciliatus CBS 366.74. The in-house enzymes were produced by growing the fungus on mono and composite substrates of cocoa pod husk (CPH) and green seaweed (GS) (Ulva lactuca sp.) with and without the addition of 25mM ammonium nitrate at 4%w/v substrate concentration in submerged condition for 144 hours. The crude enzyme extracts preparations (CEE 1-5 and CEE 1-5+AN) obtained from the fungal cultivation process were sterile-filtered and used as enzyme sources for enzymatic hydrolysis of hydrothermally pretreated corn stover using a commercial cocktail enzyme, Cellic Ctec3, as benchmark. The hydrolysis was conducted at 50ᵒC with 50mM sodium acetate buffer, pH 5 based on enzyme dosages of 5 and 10 CMCase Units/g biomass at 1%w/v dry weight substrate concentration at time points of 6, 24, and 72 hours. The enzyme activity profile of the in-house enzymes varied among the growth substrates with the composite substrates (50-75% GS and AN inclusion), yielding better enzyme activities, especially endoglucanases (0.4-0.5U/mL), β-glucosidases (0.1-0.2 U/mL), and xylanases (3-10 U/mL). However, nitrogen supplementation had no significant effect on enzyme activities of crude extracts from 100% GS substituted substrates. From the enzymatic hydrolysis, it was observed that the in-house enzymes were capable of hydrolysing the pretreated corn stover at varying degrees; however, the saccharification yield was less than 10%. Consequently, theoretical glucose yield was ten times lower than Cellic Ctec3 at both dosage levels. There was no linear correlation between glucose yield and enzyme dosage for the in-house enzymes, unlike the benchmark enzyme. It is therefore recommended that the in-house enzymes are used to complement the dosage of commercial enzymes to reduce the cost of biomass saccharification.

Keywords: enzyme production, hydrolysis yield, feedstock, enzyme blend, Polyporus ciliatus

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Authors: Salwa Karboune, Amanda Waglay

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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.

Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides

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Authors: Sudkate Chaiyo, Weena Siangproh, Orawon Chailapakul, Kurt Kalcher

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In the present work, a simple and sensitive non-enzymatic electrochemical detection of glucose in disposable paper-based sensor was developed at ionic liquid/graphene/cobalt phthalocyanine composite (IL/G/CoPc) modified electrode. The morphology of the fabricated composite was characterized and confirmed by scanning electron microscopy and UV-Vis spectroscopy. The UV-Vis spectroscopy results confirmed that the G/CoPc composite formed via the strong π–π interaction between CoPc and G. Amperometric i-t technique was used for the determination of glucose. The response of glucose was linear over the concentration ranging from 10 µM to 1.5 mM. The response time of the sensor was found as 30 s with a limit of detection of 0.64 µM (S/N=3). The fabricated sensor also exhibited its good selectivity in the presence of common interfering species. In addition, the fabricated sensor exhibited its special advantages such as low working potential, good sensitivity along with good repeatability and reproducibility for the determination of glucose.

Keywords: glucose, paper-based sensor, ionic liquid/graphene/cobalt phthalocyanine composite, electrochemical detection

Procedia PDF Downloads 164