Search results for: enzyme pre-treatment
Commenced in January 2007
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Edition: International
Paper Count: 1127

Search results for: enzyme pre-treatment

497 Ni Mixed Oxides Type-Spinel for Energy: Application in Dry Reforming of Methane for Syngas (H2 and CO) Production

Authors: Bedarnia Ishak

Abstract:

In the recent years, the dry reforming of methane has received considerable attention from an environmental view point because it consumes and eliminates two gases (CH4 and CO2) responsible for global warming by greenhouse effect. Many catalysts containing noble metal (Rh, Ru, Pd, Pt and Ir) or transition metal (Ni, Co and Fe) have been reported to be active in this reaction. Compared to noble metals, Ni-materials are cheap but very easily deactivated by coking. Ni-based mixed oxides structurally well-defined like perovskites and spinels are being studied because they possibly make solid solutions and allow to vary the composition and thus the performances properties. In this work, nano-sized nickel ferrite oxides are synthesized using three different methods: Co-precipitation (CP), hydrothermal (HT) and sol gel (SG) methods and characterized by XRD, Raman, XPS, BET, TPR, SEM-EDX and TEM-EDX. XRD patterns of all synthesized oxides showed the presence of NiFe2O4 spinel, confirmed by Raman spectroscopy. Hematite was present only in CP sample. Depending on the synthesis method, the surface area, particle size, as well as the surface Ni/Fe atomic ratio (XPS) and the behavior upon reduction varied. The materials were tested in methane dry reforming with CO2 at 1 atm and 650-800 °C. The catalytic activity of the spinel samples was not very high (XCH4 = 5-20 mol% and XCO2 = 25-40 mol %) when no pre-reduction step was carried out. A significant contribution of RWGS explained the low values of H2/CO ratio obtained. The reoxidation step of the catalyst carried out after reaction showed little amounts of coke deposition. The reducing pretreatment was particularly efficient in the case of SG (XCH4 = 80 mol% and XCO2 = 92 mol%, at 800 °C), with H2/CO > 1. In conclusion, the influence of preparation was strong for most samples and the catalytic behavior could be interpreted by considering the distribution of cations among octahedral (Oh) and tetrahedral (Td) sites as in (Ni2+1-xFe3+x) Td (Ni2+xFe3+2-x) OhO2-4 influenced the reducibility of materials and thus their catalytic performance.

Keywords: NiFe2O4, dry reforming of methane, spinel oxide, oxide zenc

Procedia PDF Downloads 268
496 Combined Treatment of Aged Rats with Donepezil and the Gingko Extract EGb 761® Enhances Learning and Memory Superiorly to Monotherapy

Authors: Linda Blümel, Bettina Bert, Jan Brosda, Heidrun Fink, Melanie Hamann

Abstract:

Age-related cognitive decline can eventually lead to dementia, the most common mental illness in elderly people and an immense challenge for patients, their families and caregivers. Cholinesterase inhibitors constitute the most commonly used antidementia prescription medication. The standardized Ginkgo biloba leaf extract EGb 761® is approved for treating age-associated cognitive impairment and has been shown to improve the quality of life in patients suffering from mild dementia. A clinical trial with 96 Alzheimer´s disease patients indicated that the combined treatment with donepezil and EGb 761® had fewer side effects than donepezil alone. In an animal model of cognitive aging, we compared the effect of combined treatment with EGb 761® or donepezil monotherapy and vehicle. We compared the effect of chronic treatment (15 days of pretreatment) with donepezil (1.5 mg/kg p. o.), EGb 761® (100 mg/kg p. o.), or the combination of the two drugs, or vehicle in 18 – 20 month old male OFA rats. Learning and memory performance were assessed by Morris water maze testing, motor behavior in an open field paradigm. In addition to chronic treatment, the substances were administered orally 30 minutes before testing. Compared to the first day and to the control group, only the combination group showed a significant reduction in latency to reach the hidden platform on the second day of testing. Moreover, from the second day of testing onwards, the donepezil, the EGb 761® and the combination group required less time to reach the hidden platform compared to the first day. The control group did not reach the same latency reduction until day three. There were no effects on motor behavior. These results suggest a superiority of the combined treatment of donepezil with EGb 761® compared to monotherapy.

Keywords: age-related cognitive decline, dementia, ginkgo biloba leaf extract EGb 761®, learning and memory, old rats

Procedia PDF Downloads 353
495 Ni Mixed Oxides Type-Spinel for Energy: Application in Dry Reforming of Methane for Syngas (H2 & Co) Production

Authors: Bouhenni Mohamed Saif El Islam

Abstract:

In the recent years, the dry reforming of methane has received considerable attention from an environmental view point because it consumes and eliminates two gases (CH4 and CO2) responsible for global warming by greenhouse effect. Many catalysts containing noble metal (Rh, Ru, Pd, Pt and Ir) or transition metal (Ni, Co and Fe) have been reported to be active in this reaction. Compared to noble metals, Ni-materials are cheap but very easily deactivated by coking. Ni-based mixed oxides structurally well-defined like perovskites and spinels are being studied because they possibly make solid solutions and allow to vary the composition and thus the performances properties. In this work, nano-sized nickel ferrite oxides are synthesized using three different methods: Co-precipitation (CP), hydrothermal (HT) and sol gel (SG) methods and characterized by XRD, Raman, XPS, BET, TPR, SEM-EDX and TEM-EDX. XRD patterns of all synthesized oxides showed the presence of NiFe2O4 spinel, confirmed by Raman spectroscopy. Hematite was present only in CP sample. Depending on the synthesis method, the surface area, particle size, as well as the surface Ni/Fe atomic ratio (XPS) and the behavior upon reduction varied. The materials were tested in methane dry reforming with CO2 at 1 atm and 650-800 °C. The catalytic activity of the spinel samples was not very high (XCH4 = 5-20 mol% and XCO2 = 25-40 mol %) when no pre-reduction step was carried out. A significant contribution of RWGS explained the low values of H2/CO ratio obtained. The reoxidation step of the catalyst carried out after reaction showed little amounts of coke deposition. The reducing pretreatment was particularly efficient in the case of SG (XCH4 = 80 mol% and XCO2 = 92 mol%, at 800 °C), with H2/CO > 1. In conclusion, the influence of preparation was strong for most samples and the catalytic behavior could be interpreted by considering the distribution of cations among octahedral (Oh) and tetrahedral (Td) sites as in (Ni2+1-xFe3+x)Td (Ni2+xFe3+2-x)OhO2-4 influenced the reducibility of materials and thus their catalytic performance.

Keywords: NiFe2O4, dry reforming of methane, spinel oxide, XCO2

Procedia PDF Downloads 370
494 Plasma Pretreatment for Improving the Durability of Antibacterial Activity of Cotton Using ZnO Nanoparticles

Authors: Sheila Shahidi, Hootan Rezaee, Abosaeed Rashidi, Mahmood Ghoranneviss

Abstract:

Plasma treatment has an explosive increase in interest and use in industrial applications as for example in medical, biomedical, automobile, electronics, semiconductor and textile industry. A lot of intensive basic research has been performed in the last decade in the field of textiles along with technical textiles. Textile manufacturers and end-users alike have been searching for ways to improve the surface properties of natural and man-made fibers. Specifically, there is a need to improve adhesion and wettability. Functional groups may be introduced onto the fiber surface by using gas plasma treatments, improving fiber surface properties without affecting the fiber’s bulk properties. In this research work, ZnO nanoparticles (ZnO-NPs) were insitue synthesized by sonochemical method at room temperature on both untreated and plasma pretreated cotton woven fabric. Oxygen and nitrogen plasmas were used for pre-functionalization of cotton fabric. And the effect of oxygen and nitrogen pre-functionalization on adhesion properties between ZnO nanoparticles and cotton surface were studied. The results show that nanoparticles with average sizes of 20-100 nm with different morphologies have been created on the surface of samples. Synthesis of ZnO-NPs was varied in the morphological transformation by changes in zinc acetate dehydrate concentration. Characterizations were carried out using Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), Inductive coupled plasma (ICP) and Spectrophotometery. The antibacterial activities of the fabrics were assessed semi-quantitatively by the colonies count method. The results show that the finished fabric demonstrated significant antibacterial activity against S. aureus in antibacterial test. The wash fastness of both untreated and plasma pretreated samples after 30 times of washing was investigated. The results showed that the parameters of plasma reactor plays very important role for improving the antibacterial durability.

Keywords: antibacterial activity, cotton, fabric, nanoparticles, plasma

Procedia PDF Downloads 522
493 Production of Keratinase and Its Insilico Characterization

Authors: Akshita Bhardwaj

Abstract:

Keratinase is an enzyme obtained from extracellular sources that is involved in biodegradation of keratin. It is a member of a group of proteases that can break down keratin into amino acids. Keratinases are produced only in the presence of substrate that contain keratin. It attacked the disulfide bond of substrate and involve in keratin degradation. Human hair, feathers, animal hard tissues, horns, claws, and hooves all contain keratin.. It exists in two form alpha keratin (found in soft tissues) and beta keratin (found in hard tissue). By taking part in the degradation of keratin, keratinases derived from microbial sources, often referred to as microbial keratinases, are important in the process of turning wastes containing keratin into products with added value. Chicken feathers contain high level of keratin protein content than other sources and became a suitable protein source. Keratinase production occurs at near alkaline pH and thermophilic temperatures. The bioprocessing of keratinous waste benefits greatly from the use of keratinases. Additionally, it lessens the issue caused by poultry excrement. The use of feather meal, along with keratinase, improves the digestion of proteins and amino acids.

Keywords: mili litre (ml), micro litre (Ul), TCA - trichloroacetic acid, OD - optical density

Procedia PDF Downloads 62
492 Hematological Profiles of Visceral Leishmaniasis Patients before and after Treatment of Anti-Leishmanial Drugs at University of Gondar Leishmania Research and Treatment Center Northwest, Ethiopia

Authors: Fitsumbrhan Tajebe, Fadil Murad, Mitikie Tigabie, Mareye Abebaw, Tadele Alemu, Sefanit Abate, Rezika Mohammedw, Arega Yeshanew, Elias Shiferaw

Abstract:

Background: Visceral leshimaniasis is a parasitic disease characterized by a systemic infection of phagocytic cells. Hematological parameters of these patients may be affected by the progress of the disease or treatment. Thus, the current study aimed to assess the hematological profiles of visceral leishmaniasis patients before and after treatment. Method: An institutional based retrospective cohort study was conducted among visceral leishmaniasis patients at University of Gondar Comprehensive Specialized Referral Hospital Leishmaniasis Research and Treatment Center from 2013 to 2018. Hematological profiles before initiation and after completion of treatment were extracted from registration book. Descriptive statics was presented using frequency and percentage. Paired t-test and Wilcoxon Signed rank test were used for comparing mean difference for normally and non- normally distributed data, respectively. Spearman and Pearson correlation analysis was used to describe the correlation of hematological parameters with different variables. P value < 0.05 was considered as statistically significant. Result: Except absolute nerutrophil count, post treatment hematological parameters show a significant increment compared to pretreatment one. The prevalence of anemia, leucopenia and thrombocytopenia was 85.5%, 83.4% and 75.8% prior to treatment and it was 58.3%, 38.2% and 19.2% after treatment, respectively. Moreover, parasite load of the disease showed statistically significant negative correlation with hematological profiles mainly with white blood cell and red blood cell. Conclusion: Majority of hematological profiles of patients with active VL have been restored after treatment, which might be associated with treatment effect on parasite proliferation and concentration of parasite in visceral organ, which directly affect hematological profiles.

Keywords: visceral leshimaniasis, hematological profile, anti-leshimanial drug, Gondar

Procedia PDF Downloads 112
491 Sniff-Camera for Imaging of Ethanol Vapor in Human Body Gases after Drinking

Authors: Toshiyuki Sato, Kenta Iitani, Koji Toma, Takahiro Arakawa, Kohji Mitsubayashi

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A 2-dimensional imaging system (Sniff-camera) for gaseous ethanol emissions from a human palm skin was constructed and demonstrated. This imaging system measures gaseous ethanol concentrations as intensities of chemiluminescence (CL) by luminol reaction induced by alcohol oxidase and luminol-hydrogen peroxide system. A conversion of ethanol distributions and concentrations to 2-dimensional CL was conducted on an enzyme-immobilized mesh substrate in a dark box, which contained a luminol solution. In order to visualize ethanol emissions from human palm skin, we developed highly sensitive and selective imaging system for transpired gaseous ethanol at sub ppm-levels. High sensitivity imaging allows us to successfully visualize the emissions dynamics of transdermal gaseous ethanol. The intensity of each pixel on the palm shows the reflection of ethanol concentrations distributions based on the metabolism of oral alcohol administration. This imaging system is significant and useful for the assessment of ethanol measurement of the palmar skin.

Keywords: sniff-camera, gas-imaging, ethanol vapor, human body gas

Procedia PDF Downloads 347
490 Effects of Palm Kernel Expeller Processing on the Ileal Populations of Lactobacilli and Escherichia Coli in Broiler Chickens

Authors: B. Navidshad

Abstract:

The main objective of this study was to examine the effects of enzymatic treatment and shell content of palm kernel expeller (PKE) on the ileal Lactobacilli and Escherichia coli populations in broiler chickens. At the finisher phase, one hundred male broiler chickens (Cobb-500) were fed a control diet or the diets containing 200 g/kg of normal PKE (70 g/kg shell), low shell PKE (30 g/kg shell), enzymatic treated PKE or low shell-enzymatic treated PKE. The quantitative real-time PCR were used to determine the ileal bacteria populations. The lowest ileal Lactobacilli population was found in the chickens fed the low shell PKE diet. Dietary normal PKE or low shell-enzymatic treated PKE decreased the Escherichia coli population compared to the control diet. The results suggested that PKE could be included up to 200 g/kg in the finisher diet, however, any screening practice to reduce the shell content of PKE without enzymatic degradation of β-mannan, decrease ileal Lactobacilli population.

Keywords: palm kernel expeller, exogenous enzyme, shell content, ileum bacteria, broiler chickens

Procedia PDF Downloads 337
489 Achievement of High L-Cysteine Yield from Enzymatic Conversion Using Eutectic Mixtures of the Substrate ATC

Authors: Deokyeong Choe, Sung Hun Youn, Younggon Kim, Chul Soo Shin

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L-Cysteine, a sulfur-containing amino acid, has been often used in the pharmaceutical, cosmetic, food, and feed additive industries. This amino acid has been usually produced by acid-hydrolysis of human hair and poultry feathers. There are many problems, such as avoidance for use of animal hair, low yields, and formation of harmful waste material. As an alternative, the enzymatic conversion of D, L-2-amino-Δ2-thiazoline-4-carboxylic acid (ATC) to L-cysteine has been developed as an environmental-friendly method. However, the substrate solubility was too low to be used in industry. In this study, high concentrations of eutectic substrate solutions were prepared to solve the problem. Eutectic melting occurred at 39°C after mixing ATC and malonic acid at a molar ratio of 1:1. The characteristics of eutectic mixtures were analyzed by FE-SEM, EDS mapping, and XPS. However, since sorbitol, MnSO4, and NaOH should be added as supplements to the substrate mixture for the activation and stabilization of the enzyme, strategies for sequential addition of total five compounds, ATC, malonic acid, sorbitol, MnSO4, and NaOH were established. As a result, eutectic substrate mixtures of 670 mM ATC were successfully formulated. After 6 h of enzymatic reaction, 550 mM L-cysteine was made.

Keywords: D, L-2-amino-Δ2-thiazoline-4-carboxylicacid, enzymatic conversion, eutectic solution, l-cysteine

Procedia PDF Downloads 414
488 Semi-pilot Biooxidation of Refractory Sulfide-Gold Ore Using Ferroplasma Acidophilum: D-(+)-Sucsore as a Booster and Columns Tests

Authors: Mohammad Hossein Karimi Darvanjooghi, Sara Magdouli, Satinder Kaur Brar

Abstract:

It has been reported that the microorganism’s attachment to the surfaces of ore samples is a key factor that influences the biooxidation in pretreatment for recovery of gold in sulfide-bearing ores. In this research, the implementation of D-(+)-Sucrose on the biooxidation of ore samples were studied in a semi-pilot experiment. The experiments were carried out in five separate jacketed columns (1 m height and 6 cm diameter) at a constant temperature of 37.5 ̊C and saturated humidity. The airflow rate and recycling solution flow rate were studied in the research and the optimum operating condition were reported. The ore sample (0.49 ppm gold grade) was obtained from the Hammond Reef mine site containing 15 wt.% of pyrite which included 98% of gold according to the results of micrograph images. The experiments were continued up to 100 days while air flow rates were chosen to be 0.5, 1, 1.5, 2, and 3 lit/min and the recycling solution (Containing 9K media and 0.4 wt.% D-(+)-Sucrose) flow rates were kept 5, 8, 15 ml/hr. The results indicated that the addition of D-(+)-Sucrose increased the bacterial activity due to the overproduction of extracellular polymeric substance (EPS) up to 95% and for the condition that the recycling solution and air flow rate were chosen to be 8 ml/hr and 2 lit/min, respectively, the maximum pyrite dissolution of 76% was obtained after 60 days. The results indicated that for the air flow rates of 0.5, 1, 1.5, 2, and 3 lit/min the ratio of daily pyrite dissolution per daily solution lost were found to be 0.025, 0.033, 0.031, 0.043, and 0.009 %-pyrite dissolution/ml-lost. The implementation of this microorganisms and the addition of D-(+)-Sucrose will enhance the efficiency of gold recovery through faster biooxidation process and leads to decrease in the time and energy of operation toward desired target; however, still other parameters including particle size distribution, agglomeration, aeration design, chemistry of recycling solution need to be controlled and monitored for reaching the optimum condition.

Keywords: column tests, biooxidation, gold recovery, Ferroplasma acidophilum, optimization

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487 Antimicrobial, Antioxidant and Enzyme Activities of Geosmithia pallida (KU693285): A Fungal Endophyte Associated with Brucea mollis Wall Ex. Kurz, an Endangered and Medicinal Plant of N. E. India

Authors: Deepanwita Deka, Dhruva Kumar Jha

Abstract:

Endophytes are the microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects. Endophytes are rich source of therapeutic substances like antimicrobial, anticancerous, herbicidal, insecticidal, immunomodulatory compounds. Brucea mollis, commonly known as Quinine in Assam, belonging to the family Simaroubaceae, is a shrub or small tree, recorded as endangered species in North East India by CAMP survey in 2003. It is traditionally being used as antimalarial and antimicrobial agent and has antiplasmodial, cytotoxic, anticancer, diuretic, cardiovascular effect etc. Being endangered and medicinal; this plant may host certain noble endophytes which need to be studied in depth. The aim of the present study was isolation and identification of potent endophytic fungi from Brucea mollis, an endangered medicinal plant, to protect it from extinction due to over use for medicinal purposes. Aseptically collected leaves, barks and roots samples of healthy plants were washed and cut into a total of 648 segments of about 2 cm long and 0.5 cm broad with sterile knife, comprising 216 segments each from leaves, barks and roots. These segments were surface sterilized using ethanol, mercuric chloride (HgCl2) and aqueous solution of sodium hypochlorite (NaClO). Different media viz., Czapeck-Dox-Agar (CDA, Himedia), Potato-Dextrose-Agar (PDA, Himedia), Malt Extract Agar (MEA, Himedia), Sabourad Dextrose Agar (SDA, Himedia), V8 juice agar, nutrient agar and water agar media and media amended with plant extracts were used separately for the isolation of the endophytic fungi. A total of 11 fungal species were recovered from leaf, bark and root tissues of B. mollis. The isolates were screened for antimicrobial, antioxidant and enzymatic activities using certain protocols. Cochliobolus geniculatus was identified as the most dominant species. The mycelia sterilia (creamy white) showing highest inhibitory activity against Candida albicans (MTCC 183) was induced to sporulate using modified PDA media. The isolate was identified as Geosmithia pallida. The internal transcribed spacer of rDNA was sequenced for confirmation of the taxonomic identity of the sterile mycelia (creamy white). The internal transcribed spacer r-DNA sequence was submitted to the NCBI (KU693285) for the first time from India. G. pallida and Penicillium showed highest antioxidant activity among all the isolates. The antioxidant activity of G. pallida and Penicillium didn’t show statistically significant difference (P˃0.05). G. pallida, Cochliobolus geniculatus and P. purpurogenum respectively showed highest cellulase, amylase and protease activities. Thus, endopytic fungal isolates may be used as potential natural resource of pharmaceutical importance. The endophytic fungi, Geosmithia pallida, may be used for synthesis of pharmaceutically important natural products and consequently can replace plants hitherto used for the same purpose. This study suggests that endophytes should be investigated more aggressively to better understand the endophyte biology of B. mollis.

Keywords: Antimicrobial activity, antioxidant activity, Brucea mollis, endophytic fungi, enzyme activity, Geosmithia pallida

Procedia PDF Downloads 166
486 Molecular Docking and Synthesis of Nitrogen-Containing Bisphosphonates

Authors: S. Ghalem, M. Mesmoudi, I. Daoudand, H. Allali

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The nitrogen-containing bisphosphonates (N-BPs) are well established as the treatments of choice for disorders of excessive bone resorption, myeloma and bone metastases, and osteoporosis. They inhibit farnesyl pyrophosphate synthase (FFPS), a key enzyme in the mevalonate pathway, resulting in inhibition of the prenylation of small GTP-binding proteins in osteoclasts and disruption of their cytoskeleton, adhesion/spreading, and invasion of cancer cells. A very few examples for synthesis of α-amino bisphosphonates based on several amino acids are known from the literature. In the present work, esters of aminoacid react with ketophsophonate (or their analog acid or acyl) to afford the desired products, α-iminophosphonates. The reaction of imine with dimethyl phosphate in the presence of catalytic amount of I2 give ester of α-aminobisphosphonate as sole product in good yield. Finally, we used computational docking methods to predict how several α-aminobisphosphonates bind to FPPS and how R and X influence. Pamidronate, β-aminobisphosphonate already marketed, was used as reference. These results are of interest since they represent a new and simple way to sythesize α-aminobisphosphonates with a free COOH group increased by R2 functionalisable and opening up the possibility of using the molecular docking to facilitate the design of other, novel FFPS inhibitors.

Keywords: drug research, cancer, α-amino bisphosphonates, molecular docking

Procedia PDF Downloads 254
485 Enhanced Enzymes Production through Immobilization of Filamentous Fungi

Authors: Zhanara B. Suleimenova, Zhazira K. Saduyeva

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Filamentous fungi are major producers of enzymes that have important applications in the food and beverage industries. The overall objective of this research is a strain improvement technology for efficient industrial enzymes production. The new way of filamentous fungi cultivation method has been developed. Such technology prolong producers’ cultivation period up to 60 days and create the opportunity to obtain enzymes repeatedly in every 2-3 days of fungal cultivation. This method is based on immobilizing enzymes producers with solid support in submerged conditions of growth. Immobilizing has a range of advantages: Decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, ability of various enzymes simultaneous production, etc. Design of proposed technology gives the opportunity to increase the activity of immobilized cells culture filtrate comparing to free cells, growing in periodic culture conditions. Thus, proposed research focuses on new, more versatile, microorganisms capable of squeezing more end-products as well as proposed cultivation technology led to increased enzymatic productivity by several times.

Keywords: filamentous fungi, immobilization, industrial enzymes production, strain improvement

Procedia PDF Downloads 347
484 The Potential Use of Flavin Mononucleotide for Photoluminescent and Bioluminescent Textile

Authors: Sweta Iyer, Nemeshwaree Behary, Jinping Guan, Guoqiang Chen, Vincent Nierstrasz

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Flavin mononucleotide widely known as 'FMN' is a biobased resource derived from riboflavin. The isoalloxazine ring present in the FMN molecule attributes the photoluminescence phenomenon, whereas FMN molecule in the presence of bacterial luciferase enzyme and co-factors such as NADH, long chain aldehyde leads to bioluminescence reaction. In this study, the FMN molecule was treated on cellulosic textile using chromojet technique and the photoluminescence property was characterized using spectroscopy technique. Further, the FMN was used as a substrate along with enzymes and co-factors to treat the non-woven textile, and the bioluminescence property was explored using luminometer equipment. The investigation revealed photoluminescence property on cellulosic textile, and the emission peak was observed at a wavelength around 530 nm with an average corrected spectral intensity of 10×106 CPS/Microamps. In addition, the measurement of nonwoven textile using bioluminescence reaction system exhibited light intensity measured in the form of relative light units (RLU). The study enabled to explore the use of FMN as both photoluminescent and bioluminescent textile. Further investigation would require for stability study of the same to provide an eco-efficient approach to obtain luminescent textile.

Keywords: flavin mononucleotide, photoluminescence, bioluminescence, luminescent textile

Procedia PDF Downloads 263
483 Bioaccessible Phenolics, Phenolic Bioaccessibilities and Antioxidant Activities of Cookies Supplemented with Pumpkin Flour

Authors: Emine Aydin, Duygu Gocmen

Abstract:

In this study, pumpkin flours (PFs) were used to replace wheat flour in cookie formulation at three different levels (10%, 20% and 30% w/w). For this purpose PFs produced by two different applications (with or without metabisulfite pre-treatment) and then dried in freeze dryer. Control sample included no PFs. The total phenolic contents of the cookies supplemented with PFs were higher than that of control and gradually increased in total phenolic contents of cookies with increasing PF supplementation levels. Phenolic content makes also significant contribution on nutritional excellence of the developed cookies. Pre-treatment with metabisulfite (MS) had a positive effect on free, bound and total phenolics of cookies which are supplemented with various levels of MS-PF. This is due to a protective effect of metabisulfite pretreatment for phenolic compounds in the pumpkin flour. Phenolic antioxidants may act and absorb in a different way in humans and thus their antioxidant and health effects will be changed accordingly. In the present study phenolics’ bioavailability of cookies was investigated in order to assess PF as sources of accessible phenolics. The content of bioaccessible phenolics and phenolic bioaccessibility of cookies supplemented with PFs had higher than those of control sample. Cookies enriched with 30% MS-PF had the highest bioaccessible phenolics (597.86 mg GAE 100g-1) and phenolic bioaccessibility (41.71%). MS application in PF production caused a significant increase in phenolic bioaccessibility of cookies. According to all assay (ABTS, CUPRAC, FRAP and DPPH), antioxidant activities of cookies with PFs higher than that of control cookie. It was also observed that the cookies supplemented with MS-PF had significantly higher antioxidant activities than those of cookies including PF. In presented study, antioxidative bioaccessibilities of cookies were also determined. The cookies with PFs had significantly (p ≤ 0.05) higher antioxidative bioaccessibilities than control ones. Increasing PFs levels enhanced antioxidative bioaccessibilities of cookies. As a result, PFs addition improved the nutritional and functional properties of cookie by causing increase in antioxidant activity, total phenolic content, bioaccessible phenolics and phenolic bioaccessibilities.

Keywords: phenolic compounds, antioxidant activity, dietary fiber, pumpkin, freeze drying, cookie

Procedia PDF Downloads 246
482 Evaluation of the Effects of Lead on Some Physiological and Hormonal Biomarkeurs among Workers

Authors: Mansouri Ouarda, Adbdennour Cherif, Boukarma Ziad

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Environmental and biological monitoring are used for the evaluation of exposure to industrial chemicals, and provide a tool for assessing workers’ exposure to chemicals. The organs or tissues where the first biological effects can be observed with increasing amounts of lead toxicity. This study aims at evaluating the effect of the metal element-trace; lead, on the sex hormones in male workers, exposed to this metal on the level of the manufacturing plant of lead accumulators. The results indicate a significant reduction of the testosterone concentration in exposed workers compared to the control. However, the rate of LH was strongly increased at the individuals exposed to Pb. A significant difference concerning the rate of FSH, the hormone Prolactin and cortisol was recorded. The indicators of the lead poisoning indicate a very highly significant increase in the value of Pbs which vary between (142-796 µg/L) among which 50% of the workers present a high lead poisoning and the value of PPZ which vary between (43-554µg/L). The biochemical parameters show a significant increase in the rate of the créatinine, the urea and the acid urique. The hepatic results show no significant differentiation in the rate of TGO and TGP between both groups of study. However the rates of the enzyme phosphatase alkaline, triglyceride, and cholesterol a significant difference were registered.

Keywords: hormons, parameters, physilogical, Pbs, PPZ

Procedia PDF Downloads 366
481 Phytochemical Analysis and in vitro Biological Activities of an Ethyl Acetate Extract from the Peel of Punica granatum L. var. Dente di Cavallo

Authors: Silvia Di Giacomo, Marcello Locatelli, Simone Carradori, Francesco Cacciagrano, Chiara Toniolo, Gabriela Mazzanti, Luisa Mannina, Stefania Cesa, Antonella Di Sotto

Abstract:

Hyperglycemia represents the main pathogenic factor in the development of diabetes complications and has been found associated with mitochondrial dysfunction and oxidative stress, which in turn increase cell dysfunction. Therefore, counteract oxidative species appears to be a suitable strategy for preventing the hyperglycemia-induce cell damage and support the pharmacotherapy of diabetes and metabolic diseases. Antidiabetic potential of many food sources has been linked to the presence of polyphenolic metabolites, particularly flavonoids such as quercetin and its glycosylated form rutin. In line with this evidence, in the present study, we assayed the potential anti-hyperglycemic activity of an ethyl acetate extract from the peel of Punica granatum L. var. Dente di Cavallo (PGE), a fruit well known to traditional medicine for the beneficial properties of its edible juice. The effect of the extract on the glucidic metabolism has been evaluated by assessing its ability to inhibit α-amylase and α-glucosidase, two digestive enzymes responsible for the hydrolysis of dietary carbohydrates: their inhibition can delay the carbohydrate digestion and reduce glucose absorption, thus representing an important strategy for the management of hyperglycemia. Also, the PGE ability to block the release of advanced glycated end-products (AGEs), whose accumulation is known to be responsible for diabetic vascular complications, was studied. The iron-reducing and chelating activities, which are the primary mechanisms by which AGE inhibitors stop their metal-catalyzed formation, were evaluated as possible antioxidant mechanisms. At last, the phenolic content of PGE was characterized by chromatographic and spectrophotometric methods. Our results displayed the ability of PGE to inhibit α-amylase enzyme with a similar potency to the positive control: the IC₅₀ values were 52.2 (CL 27.7 - 101.2) µg/ml and 35.6 (CL 22.8 - 55.5) µg/ml for acarbose and PGE, respectively. PGE also inhibited the α-glucosidase enzyme with about a 25 higher potency than the positive controls of acarbose and quercetin. Furthermore, the extract exhibited ferrous and ferric ion chelating ability, with a maximum effect of 82.1% and 80.6% at a concentration of 250 µg/ml respectively, and reducing properties, reaching the maximum effect of 80.5% at a concentration of 10 µg/ml. At last, PGE was found able to inhibit the AGE production (maximum inhibition of 82.2% at the concentration of 1000 µg/ml), although with lower potency with respect to the positive control rutin. The phytochemical analysis of PGE displayed the presence of high levels of total polyphenols, tannins, and flavonoids, among which ellagic acid, gallic acid and catechin were identified. Altogether these data highlight the ability of PGE to control the carbohydrate metabolism at different levels, both by inhibiting the metabolic enzymes and by affecting the AGE formation likely by chelating mechanisms. It is also noteworthy that peel from pomegranate, although being a waste of juice production, can be reviewed as a nutraceutical source. In conclusion, present results suggest the possible role of PGE as a remedy for preventing hyperglycemia complications and encourage further in vivo studies.

Keywords: anti-hyperglycemic activity, antioxidant properties, nutraceuticals, polyphenols, pomegranate

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480 Evaluating Antifungal Potential of Respiratory Inhibitors against Phyto-Pathogenic Fungi

Authors: Sehrish Iftikhar, Ahmad Ali Shahid, Kiran Nawaz, Waheed Anwar

Abstract:

Discovery and development of new compounds require intense studies in chemistry, biochemistry. Numerous experiments under laboratory-, greenhouse- and field conditions can be performed to select suitable candidates and to understand their full potential. Novel fungicides are fundamental to combat plant diseases. Fusarium solani is important plant pathogen. New broad spectrum foliar fungicides against complex II were designed in this study. Complex II, namely succinate dehydrogenase (SDH), or succinate quinone oxidoreductase (SQR) is a multi-subunit enzyme at the crossroads of TCA and ETC at the inner mitochondrial membrane. The need for new and innovative fungicides is driven by resistance management, regulatory hurdles and increasing customer expectations amongst others. Fungicidal activity was assessed for the effect on mycelial growth and spore germination of the fungi using fungicide amended media assay. In mycelial growth assay compounds C10 and C6 were highly active against all the isolates. The compounds C1 and C10 were found most potent in spore germination test. It fully proved that the SDHIs designed in this paper displayed as good inhibitory effects against Fusarium solani.

Keywords: Wilt, Fusarium, SDH, antifungal

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479 Hepatitis E among Pregnant Women in Urmia, Iran

Authors: Zakieh Rostamzadeh Khameneh, Nariman Sepehrvand, Khalkhali-Zahra Shirmohamadi

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Background: Although the hepatitis E virus mostly causes a self-limited disease in general population, the disease is more severe in pregnant women. Hepatitis E accounts for about 10% of pregnancy-associated deaths in southern Asia. Methods: 136 pregnant women who referred to urban health centers of Urmia for pursuing pregnancy-related health services were selected randomly and enrolled in a descriptive, cross-sectional study. Each subject was tested for the presence of anti-HEV IgG antibody using an enzyme-linked immunosorbent assay (ELISA, Dia.Pro). Results: The mean age among 136 pregnant women was 25.12±4.91 years old (range of 14-39 years). Only five cases (3.6%) among all 136 subjects were demonstrated to be seropositive for anti-HEV IgG using ELISA method. There was no significant difference between age (P=0.88), income level (P=0.19) of two seropositive and seronegative groups. All seropositive cases were from urban areas. Conclusion: The seroprevalence of anti-HEV IgG is low in the population of pregnant women in Urmia, Iran. Because of limited sample size in this study, we recommend to perform further studies with larger sample size in other regions of Iran in order to be able to systematically generalize the findings of studies to the population of Iranian pregnant women.

Keywords: pregnancy, hepatitis E, women, ELISA

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478 Detection of Total Aflatoxin in Flour of Wheat and Maize Samples in Albania Using ELISA

Authors: Aferdita Dinaku, Jonida Canaj

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Aflatoxins are potentially toxic metabolites produced by certain kinds of fungi (molds) that are found naturally all over the world; they can contaminate food crops and pose a serious health threat to humans by mutagenic and carcinogenic effects. Several types of aflatoxin (14 or more) occur in nature. In Albanian nutrition, cereals (especially wheat and corn) are common ingredients in some traditional meals. This study aimed to investigate the presence of aflatoxins in the flour of wheat and maize that are consumed in Albania’s markets. The samples were collected randomly in different markets in Albania and detected by the ELISA method, measured in 450 nm. The concentration of total aflatoxins was analyzed by enzyme-linked immunosorbent assay (ELISA), and they were ranged between 0.05-1.09 ppb. However, the screened mycotoxin levels in the samples were lower than the maximum permissible limits of European Commission No 1881/2006 (4 μg/kg). The linearity of calibration curves was good for total aflatoxins (B1, B2, G1, G2, M1) (R²=0.99) in the concentration range 0.005-4.05 ppb. The samples were analyzed in two replicated measurements and for each sample, the standard deviation (statistical parameter) is calculated. The results showed that the flour samples are safe, but the necessity of performing such tests is necessary.

Keywords: aflatoxins, ELISA technique, food contamination, flour

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477 Biomimetic Luminescent Textile Using Biobased Products

Authors: Sweta Iyer, Nemeshwaree Behary, Vincent Nierstrasz

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Various organisms involve bioluminescence for their particular biological function. The bio-based molecules responsible for bioluminescence vary from one species to another, research has been done to identify the chemistry and different mechanisms involved in light production in living organisms. The light emitting chemical systems such as firefly and bacterial luminous mostly involves enzyme-catalyzed reactions and is widely used for ATP measurement, bioluminescence imaging, environmental biosensors etc. Our strategy is to design bioluminescent textiles using such bioluminescent systems. Hence, a detailed literature work was carried out to study on how to mimic bioluminescence effect seen in nature. Reaction mechanisms in various bioluminescent living organisms were studied and the components or molecules responsible for luminescence were identified. However, the challenge is to obtain the same effect on textiles by immobilizing enzymes responsible for light creation. Another challenge is also to regenerate substrates involved in the reaction system to create a longer lasting illumination in bioluminescent textiles. Natural film-forming polymers were used to immobilize the reactive components including enzymes on textile materials to design a biomimetic luminescent textile.

Keywords: bioluminescence, biomimetic, immobilize, luminescent textile

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476 Association of Lipoprotein Lipase Gene (HindIII rs320) Polymorphisms with Moderate Hypertriglyceridemia Secondary to Metabolic Syndrome

Authors: Meryem Abi-Ayad, Biagio Arcidiacono, Eusebio Chiefari, Daniela Foti, Mohamed Benyoucef, Antonio Brunetti

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Lipoprotein Lipase (LPL) is a key enzyme for lipid metabolism; its genetic polymorphism can be a candidate for modulating lipids parameters in metabolic syndrome. The objective of the present study was to determine whether lipoproteins lipase polymorphisMetS (LPL-HindIII) could be associated with moderate hypertriglyceridemia (secondary to metabolism syndrome). The polymorphism Hind III (rs320) was assessed by PCR-RFLP in 51 MetS patients and 17 healthy controls from the hospital in Tlemcen. The logistic regression analyses showed no significant association with Hind III genotype and hypertriglyceridemia (TG ≥ 1,5g/l or TG lower treatment) (P=0,455), metabolic syndrome (P=0,455), hypertension (P=0,802) and type 2 diabetes (P=0,144). In terms of plasma biomarkers, although not statistically significant, there was a difference in TG levels (P > 0,05), which was lowest among carriers of the homogenous mutant allele (H-). In this study, there was no association between the rare allele (H-) and disease protection, and between the frequent allele (H+) and disease prevalence (hypertriglyceridemia, metabolic syndrome, hypertension, type 2 diabetes).

Keywords: moderate secondary hypertriglyceridemia, metabolic syndrome, lipids, polymorphism lipoprotein lipase, HindIII(rs320)

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475 Synthesis, Inhibitory Activity, and Molecular Modelling of 2-Hydroxy-3-Oxo-3-Phenylpropionate Derivatives as HIV-1-Integrase Inhibitors

Authors: O. J. Jesumoroti, Faridoon, R. Klein, K. A. Iobb, D. Mnkadhla, H. C. Hoppe, P. T. Kaye

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The 1, 3-aryl diketo acids (DKA) based agents represent an important class of HIV integrase (IN) strand transfer inhibitors. In other to study the chelating role of the divalent metal ion in the inhibition of IN strand transfer, we designed and synthesized a series of 2-hydroxy-3-oxo-3-phenyl propionate derivatives with the notion that such compounds could interact with the divalent ion in the active site of IN. The synthetic sequence to the desired compounds involves the concept of Doebner knoevenagel condensation, Fischer esterification and ketohydroxylation using neuclophilic re-oxidant; compounds were characterized by their IR, IHNMR, 13CNMR, HRMS spectroscopic data and melting point determination. Also, molecular docking was employed in this study and it was revealed that there is interaction with the active site of the enzyme. However, there is disparity in the corresponding anti-HIV activity determined by the experimental bioassay. These compounds lack potency at low micromolar concentration when compared to the results of the docking studies. Nevertheless, the results of the study suggest modification of the aryl ring with one or two hydroxyl groups to improve the inhibitory activity.

Keywords: anti-HIV-1 integrase, ketohydroxylation, molecular docking, propionate derivatives

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474 Effects of Different Thermal Processing Routes and Their Parameters on the Formation of Voids in PA6 Bonded Aluminum Joints

Authors: Muhammad Irfan, Guillermo Requena, Jan Haubrich

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Adhesively bonded aluminum joints are common in automotive and aircraft industries and are one of the enablers of lightweight construction to minimize the carbon emissions during transportation for a sustainable life. This study is focused on the effects of two thermal processing routes, i.e., by direct and induction heating, and their parameters on void formation in PA6 bonded aluminum EN-AW6082 joints. The joints were characterized microanalytically as well as by lap shear experiments. The aging resistance of the joints was studied by accelerated aging tests at 80°C hot water. It was found that the processing of single lap joints by direct heating in a convection oven causes the formation of a large number of voids in the bond line. The formation of voids in the convection oven was due to longer processing times and was independent of any surface pretreatments of the metal as well as the processing temperature. However, when processing at low temperatures, a large number of small-sized voids were observed under the optical microscope, and they were larger in size but reduced in numbers at higher temperatures. An induction heating process was developed, which not only successfully reduced or eliminated the voids in PA6 bonded joints but also reduced the processing times for joining significantly. Consistent with the trend in direct heating, longer processing times and higher temperatures in induction heating also led to an increased formation of voids in the bond line. Subsequent single lap shear tests revealed that the increasing void contents led to a 21% reduction in lap shear strengths (i.e., from ~47 MPa for induction heating to ~37 MPa for direct heating). Also, there was a 17% reduction in lap shear strengths when the consolidation temperature was raised from 220˚C to 300˚C during induction heating. However, below a certain threshold of void contents, there was no observable effect on the lap shear strengths as well as on hydrothermal aging resistance of the joints consolidated by the induction heating process.

Keywords: adhesive, aluminium, convection oven, induction heating, mechanical properties, nylon6 (PA6), pretreatment, void

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473 Design and Fabrication of Optical Nanobiosensors for Detection of MicroRNAs Involved in Neurodegenerative Diseases

Authors: Mahdi Rahaie

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MicroRNAs are a novel class of small RNAs which regulate gene expression by translational repression or degradation of messenger RNAs. To produce sensitive, simple and cost-effective assays for microRNAs, detection is in urgent demand due to important role of these biomolecules in progression of human disease such as Alzheimer’s, Multiple sclerosis, and some other neurodegenerative diseases. Herein, we report several novel, sensitive and specific microRNA nanobiosensors which were designed based on colorimetric and fluorescence detection of nanoparticles and hybridization chain reaction amplification as an enzyme-free amplification. These new strategies eliminate the need for enzymatic reactions, chemical changes, separation processes and sophisticated equipment whereas less limit of detection with most specify are acceptable. The important features of these methods are high sensitivity and specificity to differentiate between perfectly matched, mismatched and non-complementary target microRNAs and also decent response in the real sample analysis with blood plasma. These nanobiosensors can clinically be used not only for the early detection of neuro diseases but also for every sickness related to miRNAs by direct detection of the plasma microRNAs in real clinical samples, without a need for sample preparation, RNA extraction and/or amplification.

Keywords: hybridization chain reaction, microRNA, nanobiosensor, neurodegenerative diseases

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472 Enhancing the Sensitivity of Antigen Based Sandwich ELISA for COVID-19 Diagnosis in Saliva Using Gold Conjugated Nanobodies

Authors: Manal Kamel, Sara Maher

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Development of sensitive non-invasive tests for detection of SARS-CoV-2 antigens is imperative to manage the extent of infection throughout the population, yet, it is still challenging. Here, we designed and optimized a sandwich enzyme-linked immunosorbent assay (ELISA) for SARS-CoV-2 S1 antigen detection in saliva. Both saliva samples and nasopharyngeal swapswere collected from 170 PCR-confirmed positive and negative cases. Gold nanoparticles (AuNPs) were conjugated with S1protein receptor binding domain (RBD) nanobodies. Recombinant S1 monoclonal antibodies (S1mAb) as primery antibody and gold conjugated nanobodies as secondary antibody were employed in sandwich ELISA. Our developed system were optimized to achieve 87.5 % sensitivity and 100% specificity for saliva samples compared to 89 % and 100% for nasopharyngeal swaps, respectively. This means that saliva could be a suitable replacement for nasopharyngeal swaps No cross reaction was detected with other corona virus antigens. These results revealed that our developed ELISAcould be establishedas a new, reliable, sensitive, and non-invasive test for diagnosis of SARS-CoV-2 infection, using the easily collected saliva samples.

Keywords: COVID 19, diagnosis, ELISA, nanobodies

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471 Development and Evaluation of Novel Diagnostic Methods for Infectious Rhinotracheitis of Cattle

Authors: Wenxiao Liu, Kun Zhang, Yongqing Li

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Bovine herpesvirus 1, a member of the genus Variellovirus of the subfamily Alphaherpesvirinae, has caused severe economic cost to the bovine industry. In this study, BoHV-1 glycerol protein gD was expressed in insect cells, and the purified gD was immunized in the Balb/C mice to generate monoclonal antibodies. Based on hybridoma cell fusion techniques, 20 monoclonal antibodies against Bovine herpesvirus 1 have been obtained. Further, mAb 3F8 with neutralizing activity and gD were applied to develop a blocking enzyme-linked immunosorbent assay (Elisa) for detecting neutralizing antibodies against BoHV-1, which shows a significant correlation between the blocking Elisa and VNT. The sensitivity and specificity of the test were estimated to be 94.59% and 93.42%, respectively. Furthermore, antibody pairing tests revealed that mAb 1B6 conjugated to fluorescence microspheres was used as the capture antibody, and mAb 3F9 was used as the detectable antibody to establish the immunochromatographic assay (ICS). The ICS was conducted to detect BoHV-1 in bovine samples with high sensitivity, specificity, and good stability. Clinical sample testing revealed that the results of ICS and real-time PCR have a coincidence rate of 95.42%. Our research confirmed that the ICS is a rapid and reliable method for the diagnosis of BoHV-1. In conclusion, our results lay a solid foundation for the prevention and control of BoHV-1 infection.

Keywords: bovine disease, BoHV-1, ELISA, ICS assay

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470 Green Extraction Processes for the Recovery of Polyphenols from Solid Wastes of Olive Oil Industry

Authors: Theodora-Venetia Missirli, Konstantina Kyriakopoulou, Magdalini Krokida

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Olive mill solid waste is an olive oil mill industry by-product with high phenolic, lipid and organic acid concentrations that can be used as a low cost source of natural antioxidants. In this study, extracts of Olea europaea (olive tree) solid olive mill waste (SOMW) were evaluated in terms of their antiradical activity and total phenolic compounds concentrations, such as oleuropein, hydroxytyrosol etc. SOMW samples were subjected to drying prior to extraction as a pretreatment step. Two drying processes, accelerated solar drying (ASD) and air-drying (AD) (at 35, 50, 70°C constant air velocity of 1 m/s), were applied. Subsequently, three different extraction methods were employed to recover extracts from untreated and dried SOMW samples. The methods include the green Microwave Assisted (MAE) and Ultrasound Assisted Extraction (UAE) and the conventional Soxhlet extraction (SE), using water and methanol as solvents. The efficiency and selectivity of the processes were evaluated in terms of extraction yield. The antioxidant activity (AAR) and the total phenolic content (TPC) of the extracts were evaluated using the DPPH assay and the Folin-Ciocalteu method, respectively. The results showed that bioactive content was significantly affected by the extraction technique and the solvent. Specifically, untreated SOMW samples showed higher performance in the yield for all solvents and higher antioxidant potential and phenolic content in the case of water. UAE extraction method showed greater extraction yields than the MAE method for both untreated and dried leaves regardless of the solvent used. The use of ultrasound and microwave assisted extraction in combination with industrially applied drying methods, such as air and solar drying, was feasible and effective for the recovery of bioactive compounds.

Keywords: antioxidant potential, drying treatment, olive mill pomace, microwave assisted extraction, ultrasound assisted extraction

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469 Comparison of Storage Facilities on Different Varieties of Orange Fleshed Sweet Potato Grown in Rwanda

Authors: Jean Paul Hategekimana, Dukuzumuremyi Yvonne, Mukeshimana Marthe, Alexandre Niyonshima

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Sweet potato (Ipomoea batatas) is a very important staple food crop in Rwanda due to its high growth and consumption in all parts of the country. The effect of seven different storage conditions on the quality and nutritional composition of the three most grown and consumed varieties of orange-fleshed sweet potato (OFSP), namely Kabode, Terimbere, and Vita, were studied over a period of six weeks at Postharvest Service and Training Center of University Rwanda, Busogo Campus. The potato stored under the following conditions (zero energy cooling chamber, ground washed sweet potato, ground unwashed sweet potato, perforated washed sweet potato, perforated unwashed sweet potato, non-perforated washed sweet potato, and non-perforated unwashed sweet potato) were assessed in this study. These storage conditions are the modifications of existing methods currently used in Rwanda for suitable local climatic conditions. Hence, 30kgs of freshly harvested OFSP for each variety were bought from farmers of Gakenke and Rulindo districts and then transported to the postharvest training and service center UR-CAVM, Busogo Campus. 2.5kg of each potato sample was selected and stored under the above-mentioned storage conditions after pretreatment. Data were collected for six weeks on percent weight loss, shrinkability and the general appearance at interval of three days. The stored samples were also analyzed for moisture, crude ash, crude fiber, and reduced sugar levels during the entire storage period. Results showed the difference among the various storage conditions. It was shown that ZECC and non-perforated sacs (in the open air) storage techniques had good potential for storage of orange flesh sweet potato for up to six weeks without considerable change in physical and nutritional parameters compared to other considered conditions and, therefore, can be recommended as more useful for OSFP at farm level and during transport and market storage.

Keywords: ZECC, orange fleshed sweet potato, perforated sacs, storage conditions

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468 Screening and Improved Production of an Extracellular β-Fructofuranosidase from Bacillus Sp

Authors: Lynette Lincoln, Sunil S. More

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With the rising demand of sugar used today, it is proposed that world sugar is expected to escalate up to 203 million tonnes by 2021. Hydrolysis of sucrose (table sugar) into glucose and fructose equimolar mixture is catalyzed by β-D-fructofuranoside fructohydrolase (EC 3.2.1.26), commonly called as invertase. For fluid filled center in chocolates, preparation of artificial honey, as a sweetener and especially to ensure that food stuffs remain fresh, moist and soft for longer spans invertase is applied widely and is extensively being used. From an industrial perspective, properties such as increased solubility, osmotic pressure and prevention of crystallization of sugar in food products are highly desired. Screening for invertase does not involve plate assay/qualitative test to determine the enzyme production. In this study, we use a three-step screening strategy for identification of a novel bacterial isolate from soil which is positive for invertase production. The primary step was serial dilution of soil collected from sugarcane fields (black soil, Maddur region of Mandya district, Karnataka, India) was grown on a Czapek-Dox medium (pH 5.0) containing sucrose as the sole C-source. Only colonies with the capability to utilize/breakdown sucrose exhibited growth. Bacterial isolates released invertase in order to take up sucrose, splitting the disaccharide into simple sugars. Secondly, invertase activity was determined from cell free extract by measuring the glucose released in the medium at 540 nm. Morphological observation of the most potent bacteria was examined by several identification tests using Bergey’s manual, which enabled us to know the genus of the isolate to be Bacillus. Furthermore, this potent bacterial colony was subjected to 16S rDNA PCR amplification and a single discrete PCR amplicon band of 1500 bp was observed. The 16S rDNA sequence was used to carry out BLAST alignment search tool of NCBI Genbank database to obtain maximum identity score of sequence. Molecular sequencing and identification was performed by Xcelris Labs Ltd. (Ahmedabad, India). The colony was identified as Bacillus sp. BAB-3434, indicating to be the first novel strain for extracellular invertase production. Molasses, a by-product of the sugarcane industry is a dark viscous liquid obtained upon crystallization of sugar. An enhanced invertase production and optimization studies were carried out by one-factor-at-a-time approach. Crucial parameters such as time course (24 h), pH (6.0), temperature (45 °C), inoculum size (2% v/v), N-source (yeast extract, 0.2% w/v) and C-source (molasses, 4% v/v) were found to be optimum demonstrating an increased yield. The findings of this study reveal a simple screening method of an extracellular invertase from a rapidly growing Bacillus sp., and selection of best factors that elevate enzyme activity especially utilization of molasses which served as an ideal substrate and also as C-source, results in a cost-effective production under submerged conditions. The invert mixture could be a replacement for table sugar which is an economic advantage and reduce the tedious work of sugar growers. On-going studies involve purification of extracellular invertase and determination of transfructosylating activity as at high concentration of sucrose, invertase produces fructooligosaccharides (FOS) which possesses probiotic properties.

Keywords: Bacillus sp., invertase, molasses, screening, submerged fermentation

Procedia PDF Downloads 221