1,721,142 research outputs found
Nonhydrolytic sol-gel synthesized oligosiloxane resin reinforced thiol-ene photocured coatings for the immobilization of acetylcholinesterase
No full textAcetylcholinesterase (AChE), which is responsible for the hydrolysis of neurotransmitter acetylcholine, is a critical enzyme for the nervous system and also a biomarker for organophosphorous pesticide detection. The immobilization of AChE is an active area of research and recently the use of sol-gel-derived materials for enzyme immobilization has gained a lot of attraction. In this work, AChE was covalently immobilized onto a photocured substrate which was reinforced with an oligosiloxane resin. The oligosiloxane resin was designed to have both vinyl and epoxide groups and prepared via nonhydrolytic sol-gel technique. The strategy employed in this study offered a platform that has good mechanical and thermal properties and also suitable for modification. Thus, AChE was also immobilized onto these substrates after amine modification of the epoxy groups and followed by glutaraldehyde activation. Over 80% enzyme immobilization yield was achieved. At certain pH values (5.5 and 8.5) and under relatively higher temperatures (above 40 degrees C) the immobilized enzymes were found to have higher catalytic activity than the free enzyme. Furthermore, by immobilization the reuse and the storage stability of the enzyme was improved and the stability of the immobilized enzyme against the inhibitory effects of certain metal cations was enhanced [GRAPHICS] . Nonhydrolytic sol-gel synthesized oligosiloxane resin reinforced thiol-ene photocured coatings for the immobilization of acetylcholinesterase. Emrah CAKMAKCI, Serap DEMIR. HighlightsAn oligosiloxane resin was prepared via nonhydrolytic sol-gel technique.The oligosiloxane resin was used to reinforce thiol-ene photocured coatings.Acetylcholinesterase was immobilized onto the photocured coatings.By immobilization, storage stability, reuse and metal ion resistance were improved
Maleic anhydride functionalization of OSTE based coatings via thiol-ene Click reaction for the covalent immobilization of xylanase
No full textCombination of Off-Stoichiometry Thiol-Ene (OSTE) polymers with thiol-ene Click reaction is a versatile method for the development of advanced functional materials. In this study OSTE based photocurable coating materials with excess thiol groups on the surface were prepared and functionalized with maleic anhydride via thiol-ene Click reaction. Xylanase enzyme was then covalently immobilized onto these coatings. Maleic anhydride functionalization of the prepared OSTE coatings was proved by FTIR and contact angle measurements. The developed functional support was found to have high affinity for xylanase enzyme and the immobilization capacity was determined as 212 mg g(-1). Almost 100% of immobilization yield was achieved. Immobilization significantly improved the stability of the xylanase at pH values over 6.5 and temperatures exceeding 60 degrees C. Moreover the immobilized xylanase exhibited superior reusability and storage stability compared to free xylanase. (C) 2016 Elsevier B.V. All rights reserved
Physical and Covalent Immobilization of Lipase onto Amine Groups Bearing Thiol-Ene Photocured Coatings
No full textIn this study, amine groups containing thiol-ene photocurable coating material for lipase immobilization were prepared. Lipase (EC 3.1.1.3) from Candida rugosa was immobilized onto the photocured coatings by physical adsorption and glutaraldehyde-activated covalent bonding methods, respectively. The catalytic efficiency of the immobilized and free enzymes was determined for the hydrolysis of p-nitrophenyl palmitate and also for the synthesis of p-nitrophenyl linoleate. The storage stability and the reusability of the immobilized enzyme and the effect of temperature and pH on the catalytic activities were also investigated. The optimum pH for free lipase and physically immobilized lipase was determined as 7.0, while it was found as 7.5 for the covalent immobilization. After immobilization, the optimum temperature increased from 37 A degrees C (free lipase) to 50-55 A degrees C. In the end of 15 repeated cycles, covalently bounded enzyme retained 60 and 70 % of its initial activities for hydrolytic and synthetic assays, respectively. While the physically bounded enzyme retained only 56 % of its hydrolytic activity and 67 % of its synthetic activity in the same cycle period. In the case of hydrolysis V (max) values slightly decreased after immobilization. For synthetic assay, the V (max) value for the covalently immobilized lipase was found as same as free lipase while it decreased dramatically for the physically immobilized lipase. Physically immobilized enzyme was found to be superior over covalent bonding in terms of enzyme loading capacity and optimum temperature and exhibited comparable re-use values and storage stability. Thus, a fast, easy, and less laborious method for lipase immobilization was developed
alpha-Amylase immobilization on functionalized nano CaCO3 by covalent attachment
No full textIn this study, a-amylase was immobilized on glutaraldehyde activated silanized calcium carbonate nanoparticles by a using covalent binding method. The surface modified nano calcium carbonate (CaCO3) were characterized using FTIR and SEM. Immobilization yield was found as 199.43 mg/g of calcium carbonate nanoparticles. The maximum activity was observed at pH 6.5. The immobilized enzyme had a higher activity at elevated temperature (5090 degrees C) than the free one. Reuse studies demonstrated that the immobilized enzyme could reuse 25 times while retaining 18.2% of its activity. Free enzyme lost its activity completely within 15 days. Vmax values for the free and immobilized enzymes were calculated as 10 and 0.35 mg/mL/min, respectively
Magnetic nanoparticle containing thiol-ene crosslinked hydrogels for controlled and targeted release of hydrophobic drugs
No full textThis study reports a straightforward but very effective method to produce nanocomposite hydrogels for targeted drug delivery system. An organic network containing both hydrophilic and hydrophobic components was obtained using thiol-ene cross-linking. Magnetite nanoparticles (MNPs) were synthesized by chemical co-precipitation. The surface of the MNPs was modified with allyl groups to avoid agglomeration. Composite hydrogels were prepared by addition of surface modified MNPs to hydrogel formulation at different ratios. The prepared composite hydrogels were investigated for their ability to loading and release hydrophobic drugs tamoxifen and clarithromycin. The morphological and structural characterizations of MNPs were performed by Fourier transform infrared spectroscopy (FT-IR) and Environmental scanning electron microscopy (SEM) and Environmental scanning transmission electron microscopy (STEM). Moreover the superparamagnetic behavior of the hydrogels was studied by a vibrating sample magnetometer (VSM). The saturation magnetization values increased with the magnetite content. Swelling, gel content and degradation of the hydrogels were examined. According to the loading of drugs, the maximum loading efficiency was 91% for tamoxifen and 70% for clarithromycin due to the stronger hydrophobic interactions. In vitro release studies showed that controlled release of the drugs occurred over a prolonged period of time. Therefore, drug loaded hydrogels can be considered as potential long-term sustained drug release systems. POLYM. COMPOS., 39:E200-E209, 2018. (c) 2016 Society of Plastics Engineer
Immobilization of alpha-amylase onto poly(glycidyl methacrylate) grafted electrospun fibers by ATRP
No full textIn this study, novel alpha-amylase immobilized poly(vinyl alcohol) (PVA) nanofibers were prepared. The PVA nanofiber surfaces were functionalized with 2-bromoisobutyryl bromide (BiBBr) and followed by surface initiated atom transfer radical polymerization (SI-ATRP) of glycidyl methacrylate (GMA). The morphology of the poly(glycidyl methacrylate) (PGMA) grafted PVA nanofibers was characterized by scanning electron microscopy (SEM). Also PGMA brushes were confirmed by X-ray photo electron microscopy (XPS). alpha-Amylase was immobilized in a one step process onto the PGMA grafted PVA nanofiber. The characteristic properties of the immobilized and free enzymes were examined. The thermal stability of the enzyme was improved and showed maximum activity at 37 degrees C by immobilization, pH values of the maximum activity of the free and immobilized enzymes were also found at 6.0 and 6.5, respectively. Free enzyme lost its activity completely within 15 days. The immobilized enzyme lost only 23.8% of its activity within 30 days. (C) 2015 Elsevier B.V. All rights reserved
Immobilization of pectinase on polyethyleneimine based support via spontaneous amino-yne click reaction
Get PDFThe immobilization of an enzyme can improve catalytic activity, stability, and reusability of its. In this study, we investigated a new method for enzyme immobilization. Alkyne-pectinase was first immobilized on the polyethyleneimine-based cryogel via a spontaneous amino-yne click reaction under very mild conditions and then the apple juice was clarified. Amino-yne click reactions do not need any photoinitiator or catalyst, unlike other click reactions. The immobilization efficiency of the alkyne pectinase was 90%. The immobilized enzyme continued to retain 70% of its initial activity after 60 days. An improvement observed in the pH tolerance in the range of 6.5-8.0. The higher thermal tolerance of the immobilized pectinase was increased above 50 degrees C. Immobilized pectinase showed 100% activity at 55 degrees C and pH 6.5. The clarification rate of apple juice was achieved about 50% by the pectinase immobilized support. (C) 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved
Covalent immobilization of a-amylase onto thermally crosslinked electrospun PVA/PAA nanofibrous hybrid membranes
No full textPoly(vinyl alcohol)/poly(acrylic acid) (PVA/PAA) nanofibers with the fiber diameter of 100150 nanometers were fabricated by electrospinning. PVA/PAA nanofibers were crosslinked by heat-induced esterification and resulting nanofiber mats insoluble in water. a-Amylase was covalently immobilized onto the PVA/PAA nanofiber surfaces via the activation of amine groups in the presence of 1,1'-carbonyldiimidazole. The immobilized a-amylase has more resistance to temperature inactivation than that of the free form and showed maximum activity at 50 degrees C. pH-dependent activities of the free and immobilized enzymes were also investigated, and it was found that the pH of maximum activity for the free enzyme was 6.5, while for the optimal pH of the immobilized enzyme was 6.0. Reuse studies demonstrated that the immobilized enzyme could reuse 15 times while retaining 81.7% of its activity. Free enzyme lost its activity completely within 15 days. Immobilized enzyme lost only 17.1% of its activity in 30 days. (C) 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 201
Amine functional magnetic nanoparticles via waterborne thiol-ene suspension photopolymerization for antibody immobilization
No full textThe modification of magnetic nanoparticles (MNPs) via different routes for biomolecule binding is an attractive area of research. Waterborne thiol-ene suspension photopolymerization (TESP) can be a useful method for preparing functional MNPs. In this study, for the very first time waterborne TESP was performed in the presence of MNPs. Neat MNPs were coated and in situ functionalized with amine groups by using thiol-ene chemistry. Engrailed-2 (EN2) protein, a potential biomarker for various cancers such as prostate cancer, bladder cancer, breast cancer and ovarian cancer, is known to be a strong binder to a specific DNA sequence (50-TAATTA-30) to regulate transcription. Anti-EN2 antibodies were immobilized onto these MNPs by physical adsorption and covalent bonding methods, respectively. The amount of the physically immobilized antibodies (0.54 mg/g) were found to be lower than the loading of the covalently bonded antibodies (1.775 mg/g). The biomarker level in the artificial solutions prepared was determined by enzyme-linked immunosorbent assay. Coated MNPs were characterized by FTIR, TGA, SEM and STEM. After TESP, the average diameter of the neat magnetite nanoparticles increased from similar to 15 nm to similar to 32 nm
Preparation and characterization of sol-gel hybrid coating films for covalent immobilization of lipase enzyme
No full textIn this study UV-curable hybrid epoxy-silica polymer films were prepared via sol-gel method. Lipase (EC 3.1.1.3) from Candida rugosa was covalently immobilized onto hybrid epoxy-silica polymer films and immobilization capacity of polymer films was found 7.22 mg g(-1). The morphology of the polymeric support was characterized by scanning electron microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). Immobilized and free enzymes were used in two different reaction systems: hydrolysis of p-nitrophenyl palmitate in aqueous medium and synthesis of p-nitrophenyl linoleate (from p-nitrophenol and linoleic acid) in n-hexane medium. The effect of temperature on hydrolytic and synthetic activities was investigated and observed maximum activities at 50 degrees C and 45 degrees C for immobilized enzyme, orderly. Km values for free enzyme were determined 0.71 and 1.12 mM by hydrolytic and synthetic activity assays, respectively, while these values were observed as 0.91 mM and 1.19 mM for immobilized enzyme. At the end of 30 repeated cycles, 56% and 59% of initial activities remained for hydrolytic and synthetic assays, respectively. Native enzyme lost its activity completely within 20 days, whereas the immobilized enzyme retained for hydrolytic and synthetic activities was approximately 82% and 72%, respectively, under the same storage time. (C) 2016 Elsevier B.V. All rights reserved
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