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    Optimization of l-asparaginase immobilization onto calcium alginate beads

    , Article Chemical Engineering Communications ; Volume 204, Issue 2 , 2017 , Pages 216-220 ; 00986445 (ISSN) Bahraman, F ; Alemzadeh, I ; Sharif University of Technology
    Taylor and Francis Ltd  2017
    Abstract
    In this study, anti-leukemic enzyme L-asparaginase (E.C.3.5.1.1) from Escherichia coli ATCC 11303 was modified by the microencapsulation technique onto calcium alginate beads. Using response surface methodology (RSM), a three-level full factorial design, the values of concentration of sodium alginate, concentration of calcium chloride, and enzyme loading were investigated to obtain the highest residual L-asparaginase (L-ASNase) activity % (immobilized enzyme activity/free enzyme activity). The effects of the studied factors on immobilization were evaluated The predicted values by the model were close to the experimental values, indicating suitability of the model. The results presented that... 

    Investigation of intestinal enzyme activity and effects of non-starch polysaccharide on it

    , Article International Journal of Engineering and Technology(UAE) ; Volume 7, Issue 4 , 2018 , Pages 28-31 ; 2227524X (ISSN) Piryaei, M ; Motamedi, A ; Mehrabi Far, A ; Sharif University of Technology
    Science Publishing Corporation Inc  2018
    Abstract
    This experiment investigates the effects of utilizing Rahnama cultivar with high non-starchy polysaccharide content and supplementation of xylanase enzyme in poultry feed on the productivity features, nutrient digestibility and intestinal enzymes activity of 21-47 week laying hens. The experiment was conducted quite randomly and in factorial design that included eight treatments with 4 wheat levels (zero, 23, 46 and 69% that contained 1.8, 2.0, 2.2, 2.4% of xylose respectively) and two level enzymes (with and without enzyme) and 5 replications (6 hens) in each replication. During the experiment, by xylose level increase in diet, the weight (p > 0.05) and mass of the egg (p > 0.01) decreased... 

    Superparamagnetic enzyme-graphene oxide magnetic nanocomposite as an environmentally friendly biocatalyst: Synthesis and biodegradation of dye using response surface methodology

    , Article Microchemical Journal ; Volume 145 , 2019 , Pages 547-558 ; 0026265X (ISSN) Kashefi, S ; Borghei, S. M ; Mahmoodi, N. M ; Sharif University of Technology
    Elsevier Inc  2019
    Abstract
    The unique properties of graphene oxide (GO) nanosheets were integrated with the superparamagnetic characteristics of the CuFe2O4 nanoparticles to synthesize the magnetic graphene oxide (MGO), which was chemically modified with 3-amino propyl trimethoxy silane (APTMS) to functionalize the amine group on MGO (MGO-NH2). Afterward, MGO-NH2 was activated with glutaraldehyde (GLU) as a crosslinking agent to synthesize the functionalized MGO (fMGO) and its capability toward covalent Laccase immobilization was investigated. The comprehensive structural analysis using various characterization techniques, including Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), vibrating... 

    A numerical model for predicting hepatocytes ureagenesis and its related inborn enzyme deficiencies: case studies

    , Article Scientia Iranica ; Volume 26, Issue 1B , 2019 , Pages 408-420 ; 10263098 (ISSN) Sharifi, F ; Firoozabadi, B ; Saidi, M. S ; Firoozbakhsh, K ; Sharif University of Technology
    Sharif University of Technology  2019
    Abstract
    One important functionality of liver cells is ammonia detoxification and urea production. In this study, a numerical model of the urea cycle in hepatocytes was developed. Navier Stokes and convection equations were employed to study the process of ammonia elimination and urea production using a microfluidic channel. The concentration of urea and ammonia throughout the channel was obtained. Furthermore, the urea cycle was modelled with respect to its four main enzymes. This resulted in twelve rate equations that were solved to determine the concentration of each metabolite participating in the urea cycle. Application of results implied common disorders such as hyperammonemia types I and II... 

    A numerical model for predicting hepatocytes ureagenesis and its related inborn enzyme deficiencies: Case studies

    , Article Scientia Iranica ; Volume 26, Issue 1B , 2019 , Pages 408-420 ; 10263098 (ISSN) Sharifi, F ; Firoozabadi, B ; Saidi, M. S ; Firoozbakhsh, K ; Sharif University of Technology
    Sharif University of Technology  2019
    Abstract
    One important functionality of liver cells is ammonia detoxification and urea production. In this study, a numerical model of the urea cycle in hepatocytes was developed. Navier Stokes and convection equations were employed to study the process of ammonia elimination and urea production using a microfluidic channel. The concentration of urea and ammonia throughout the channel was obtained. Furthermore, the urea cycle was modelled with respect to its four main enzymes. This resulted in twelve rate equations that were solved to determine the concentration of each metabolite participating in the urea cycle. Application of results implied common disorders such as hyperammonemia types I and II... 

    Nano reengineering of horseradish peroxidase with dendritic macromolecules for stability enhancement

    , Article Enzyme and Microbial Technology ; Volume 50, Issue 1 , 2012 , Pages 10-16 ; 01410229 (ISSN) Khosravi, A ; Vossoughi, M ; Shahrokhian, S ; Alemzadeh, I ; Sharif University of Technology
    Abstract
    A simple bio-conjugation procedure to surround a single horseradish peroxidase (HRP) enzyme molecule with dendritic polyester macromolecules (polyester-32-hydroxyl-1-carboxyl bis-MPA dendron, generation 5) was proposed. The characterization of resultant nanoparticles entitled HRP dendrozyme, was performed by transmission electron microscopy, dynamic light scattering, gel permeation chromatography and Fourier transform infrared spectroscopy. The results showed that HRP nanoparticles were spherical in shape and have an average size of 14 ± 2. nm in diameter. Furthermore, bio-conformational characterization of HRP dendrozyme was performed by means of circular dichroism and fluorescence... 

    Magnetite nanoparticle as a support for stabilization of chondroitinase ABCI

    , Article Artificial Cells, Nanomedicine and Biotechnology ; Volume 47, Issue 1 , 2019 , Pages 2721-2728 ; 21691401 (ISSN) Askaripour, H ; Vossoughi, M ; Khajeh, K ; Alemzadeh, I ; Sharif University of Technology
    Taylor and Francis Ltd  2019
    Abstract
    Chondroitinase ABCI (cABCI) is a drug enzyme that can be used to treat spinal cord injuries. Due to low thermal stability of cABCI, this enzyme was immobilized on Fe3O4 nanoparticle to increase its thermal stability. The size and morphology, structure and magnetic property of the Fe3O4 nanoparticles were characterized by the analyses of SEM, XRD and VSM, respectively, and FTIR spectroscopy was employed to confirm the immobilization of cABCI on the surface of Fe3O4 nanoparticles. The results indicated that the optimum conditions for pH, temperature, cABCI-to-Fe3O4 mass ratio and incubation time in immobilization process were 6.5, 15 °C, 0.75 and 4.5 h, respectively, and about 0.037 mg cABCI... 

    Preparation of Polyelectrolyte-Coated Magnetic Silica Nanocomposite Particles and Investigation of their Application for Enzyme Immobilization and Biodiesel Production

    , Ph.D. Dissertation Sharif University of Technology Esmaeilnejad-Ahranjani, Parvaneh (Author) ; Kazemeini, Mohammad (Supervisor) ; Arpanaei, Ayyoob (Supervisor) ; Singh, Gurvinder (Co-Advisor)
    Abstract
    In this study, polyelectrolyte-coated magnetic silica nanocomposites were prepared via the covalent attachment of polyelectrolytes onto the as-prepared magnetite-silica particles with core-shell structures and average diameter of 210±50 nm, and subsequently applied for the immobilization of lipase molecules. In this regard, polyethyleneimine (PEI) with two molecular weights of 10000 and 750000 and polyacrylic acid (PAA) with two molecular weights of 1800 and 100000 were utilized. The type and molecular weight of the polyelectrolyte molecules greatly impacted the concentration of binding sites on the particles surface and consequently on the lipase immobilization efficiency and loading... 

    α-Amylase Immobilization on Nanocomposite Magnetic Silica Particles and Characterization of the Prepared Nanobiocatalysts

    , M.Sc. Thesis Sharif University of Technology Sayyahmanesh, Maryam (Author) ; Mashayekhan, Shohreh (Supervisor) ; Arpanaei, Ayyoob (Supervisor)
    Abstract
    Enzymes are widely used in industry, however, the expensive cost, low stability, and the limited activity in a certain range of temperature and pH, have limited the usage of such bio-catalists. In addition to increasing and preserving their activity under different conditions, the enzyme immobilization can improve its stability in different pH, temperature values and increase its thermostability, storage stability and the possibility of their recycling. Meanwhile, utilizing the super-magnetic systems could be very helpful as well. For instance, separation of the the magnetic nano-particles like Fe3O4 from reaction system using an external magnetic field is easily possible. In this study,... 

    Studying of Laccase Immobilization on Nanoparticles for Bisphenol A Removal

    , M.Sc. Thesis Sharif University of Technology Sadeghzadeh, Sadegh (Author) ; Borghei, Mehdi (Supervisor) ; Ghobadi Nejad, Zahra (Co-Supervisor)
    Abstract
    Bisphenol A is one of Endocrine Disturbing Chemicals. Enzymatic removal of bisphenol A is an interesting and green method. However, application of enzymes has been hampered due to lower operational stability and lots of difficulties in recovery and recycling. Enzyme immobilization can ba an innovative approach to circumvent these drawbacks. Laccase from Trametes hirsuta used for Bisphenol A removal. Firstly, magnetic nanoparticles were synthesized by coprecipitation method and aminofunctionalized by (3-aminopropyl)-mmethoxysilane. Subsequently, laccase was precipitated with i-propanol in the presence of aminofunctionalized nanoparticles and followed by addition of glutaraldehyde to the final... 

    Removal of Pharmaceuticals from Polluted Water Using Carbon Nanotube-based Membranes Coated with Enzyme

    , M.Sc. Thesis Sharif University of Technology Masjoudi, Mahsa (Author) ; Borghei, Mehdi (Supervisor) ; Ghobadi Nejad, Zahra (Co-Supervisor)
    Abstract
    Today, presence of micropollutants, including pharmaceutical compounds, in aquatic systems has turned into a major environmental challenge threatening the health of humans and many aquatic and terrestrial organisms. In the recent years, biodegradation of pharmaceuticals using enzymes, such as laccase, has been presented as a green method for eliminating these types of pollutants. Furthermore, regarding the improved stability and reusability of immobilized enzymes, these types of biocatalysts are suitable for industrial applications. Enzymatic membrane reactor is a promising method of enzymatic treatment and fabrication of a high-efficiency enzymatic membrane is a major step to begin this... 

    The Study of Function of Lipase Biocatalyst Immobilized on Silica Nanoparticles Used for Biodiesel Production

    , M.Sc. Thesis Sharif University of Technology Kalantari, Mohammad (Author) ; Kazemini, Mohammad (Supervisor) ; Arpanaei, Ayyoob (Supervisor) ; Tabandeh, Fatemeh (Co-Advisor)
    Abstract
    Superparamagnetic core/shell nonporous (S1) and core/shell/shell mesoporous (S2 and S3) nanocomposite magnetite/silica particles with a magnetite cluster core of 130±30 nm, a nonporous silica shell of 90±10 nm thickness, and a mesoporous silica shell of 70±15 nm thickness were prepared thorough a simple method. Mesoporous particles were prepared with two BJH pore sizes (2.44 for S2 and 3.76 nm for S3 particles, respectively). The fabricated S1, S2 and S3 particles present high saturation magnetization values of 20, 13 and 17 emu/g, respectively. As a biological application, the lipase from Pseudomonas cepacia was successfully immobilized onto the amino-functionalized nanocomposite particles... 

    Synthesis and Stability Evaluation of Single Enzyme Nanoparticles for Enzymatic Based Processes

    , Ph.D. Dissertation Sharif University of Technology Khosravi, Arezoo (Author) ; Vossoughi, Manoochehr (Supervisor) ; Shahrokhian, Saeed (Co-Advisor) ; Alemzadeh, Iran (Co-Advisor)
    Abstract
    Enzymes, the nanometric biocatalysts that play a key role in biochemical reactions of living cells and bio-organisms, offer various applications including biosensing, bioremediation and chemical synthesis. A simple bio-conjugation procedure to surround a single horseradish peroxidase (HRP) enzyme molecule with dendritic polyester macromolecules (Polyester-32-hydroxyl-1-carboxyl bis-MPA Dendron, Generation 5) was proposed in this project. The characterization of resultant nanoparticles entitled HRP dendrozyme, was performed by TEM, DLS, GPC and FTIR spectroscopy. The results showed that HRP nanoparticles were spherical in shape and have an average size of 14 nm in diameter. Furthermore,... 

    Comparing the Performance of Centrifuge, Membrane Filter and Filter Press for Clarification of Lipase Enzyme

    , M.Sc. Thesis Sharif University of Technology Taher Aslani, Ahmad Reza (Author) ; Rousta Azad, Reza (Supervisor)
    Abstract
    Lipases (Triacyl Glycerol Acyl Hydrolase, EC.3.1.1.3) are multidimensional biocatalysts which have many applications in food industrials, dairy, fats and oils processing, detergents, synthesis of pure chemical and pharmaceutical materials, paper industrial and cosmetics. Generally, industrial enzymes are produced extracellularly, so their separation from a low-cost complex medium (usually used for production) is one of the cost stages in their production process. Filtration is the most economical method of separation in the production of enzymes, but due to the small size of colloidal particles and cell masses present in the fermentation environment, its use is subject to limitations. In... 

    studying the Effect of Adding Nanoparticles in Polymer/salt Aqueous Two Phase Systems for Separating of Alpha-amylase Enzyme

    , Ph.D. Dissertation Sharif University of Technology Dehnavi, Mohsen (Author) ; Vossoughi, Manoochehr (Supervisor) ; Pazuki, Gholamreza (Supervisor)
    Abstract
    The effects of adding nanoparticles in partitioning of α-amylase in polymer/salt aqueous two phase system (ATPS) was studied in this research. First the binodal curves of magnesium sulfate salt with PEGs 2000,3000,4000,6000 and 10000 at 23 ° C has been plotted and the suitable system which was PEG 6000/ magnesium sulfate has been selected for studying the partitioning of enzyme. Thereupon the partition coefficient of α-amylase was measured and then effect of adding silica and graphene oxide nanoparticles was investigated. It was observed that nanoparticles caused a dramatic increase in partition coefficient and yield recovery (of top phase). For further investigation the partitioning of... 

    Experimental Study of Biological Synthesis, Stabilization and Delivery of the Recombinant Protein of Chondroitinase

    , Ph.D. Dissertation Sharif University of Technology Askaripour, Hossein (Author) ; Vossoughi, Manouchehr (Supervisor) ; Alemzadeh, Iran (Supervisor) ; Khajeh, Khosro (Supervisor)
    Abstract
    In this research, immobilization method was employed to increase the thermal stability of Chondroitinase ABC I (cABC I) enzyme. In first step, magnetite nanoparticle (Fe3O4) was selected as a support and cABC I enzyme was attached via the adsorption method. The results showed that pH=6.5, temperature 15 ˚C, enzyme-to-support mass ratio 0.75, and incubation time 4.5 hr were the appropriate conditions for immobilizing cABC I enzyme on Fe3O4 nanoparticle. It was also found that the optimum pH for free and immobilized enzymes was 7.6 and 8.0, respectively. The maximum activity of free enzyme was obtained at 25 ˚C, whereas the activity of immobilized enzyme was almost constant in the temperature... 

    Investigate Kinetic Parameters of L-Asparaginase Ii as Well as its Performance in Organic Reactions and Modifying the Enzyme Through Directed Evolution for Carrying Out Promiscuous Reactions

    , M.Sc. Thesis Sharif University of Technology Noori Hosseinabadi, Zahra (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    L-asparaginase II is an amidohydrolase which hydrolyzes L-asparagine to furnish L-aspartic acid and ammonia. The enzyme has played an important role for 40 years in treating cancer in persons with juvenile leukemias or lymphomas. Furthermore, the enzyme has become important in food industries and biosensors technology.In this study, the expression of recombinant L-asparaginase II in E. Coli was optimized and the purification of this enzyme from growth culture was carried out with nickel-charged affinity resin. Afterwards, the activity, and kinetic parameters of purified recombinant L-asparaginase II including Km, kcat, Vmax, and enzyme efficiency were measured.In the next step, random... 

    Studying Immobilization of Laccase by Entrapment and Its Applications in Micropollutants Removal from Aqueous Wastewater

    , M.Sc. Thesis Sharif University of Technology Toutounchi, Arvin (Author) ; Yaghmaei, Soheila (Supervisor) ; Ghabodinejad, Zahra (Co-Supervisor)
    Abstract
    One of the major environmental problems in recent years is the presence of micropollutants that cause many problems for the environment and living organisms. The micropollutant bisphenol A and the drug diclofenac are one of the pollutants and endocrine disruptors. Usually, conventional biological and physico-chemical methods do not have a high ability to remove such pollutants. Also, advanced oxidation methods usually lead to the production of highly toxic products. Using enzymes is an attractive method to remove these micropollutants. Laccase enzyme is one of the most famous enzymes in the field of biotherapy because it can remove a wider range of pollutants and it is one of the... 

    Immobilization of Laccase Using Metal-Organic Framework and its Application in Micropollutant Removal

    , M.Sc. Thesis Sharif University of Technology Ghassemi, Raman (Author) ; Yaghmaei, Soheila (Supervisor) ; Ghobadinejad, Zahra (Supervisor)
    Abstract
    Nowadays, micropollutants present in aquatic environments posses a significant threat to the well-being and health of human beings. Various physicochemical and advanced approaches have been proposed to confront these hazards. In recent years, biological treatment methods have gained great interest for their green approach. One of these biological treatment options is the treatment of polluted waters by enzymes. Enzymatic treatment of micropollutants can be achieved by oxidoreductase enzymes. One of the most important and widely used oxidoreductase enzymes employed for water treatment is Laccase. However, enzyme immobilization is employed to counter the drawbacks of using enzymes, such as... 

    Treatment of Triclosan Through Enhanced Microbial Biodegradation

    , M.Sc. Thesis Sharif University of Technology Ghafouri, Mahsa (Author) ; Yaghmaei, Soheila (Supervisor) ; Ghobadi Nejad, Zahra (Supervisor)
    Abstract
    Triclosan is widely used as an antibacterial agent in personal health care products, especially in recent years with the spread of the Coronavirus. This substance is one of the emerging contaminants, and due to its stable and toxic nature, this material cannot be destroyed in the process of biological wastewater treatment. In this research, a new bacterial strain was isolated and purified from industrial wastewater for the biological removal of triclosan. Based on 16S rRNA sequencing, it was found that the isolated strain belongs to the Enterobacter cloacae family and can grow in high concentrations of triclosan. It was observed that naphthalene dioxygenase and laccase enzymes are active...