Chemistry / Kimya
Permanent URI for this collectionhttps://hdl.handle.net/11147/4072
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Article Citation - WoS: 7Citation - Scopus: 8Thermoalkalophilic Recombinant Esterase Entrapment in Chitosan/Calcium Beads and Its Characterization(Wiley, 2021) Tercan, Cisem; Sürmeli, Yusuf; Şanlı Mohamed, GülşahBACKGROUND Esterases (EC 3.1.1.1), a class of hydrolases, degrade the ester bonds of lipids into alcohol and carboxylic acids and synthesize carboxylic ester bonds. They are used in a variety of biotechnological, industrial, environmental, and pharmaceutical applications due to their many valuable properties. Particularly, extremophilic esterases with many superior properties are of great interest for various reactions. Immobilization of enzymes may provide some advantages over free enzymes not only to improve the properties of enzymes but also to increase the reusability of biocatalyst in industrial applications. Therefore, many different immobilization applications for enzymes have been reported in various studies. To our knowledge, a thermophilic esterase has not so far been immobilized by entrapment using chitosan/calcium/alginate-blended beads. Here, we reported the immobilization of thermoalkalophilic recombinant esterase by entrapment using chitosan/calcium/alginate-blended beads, and then the entrapped esterase was characterized biochemically in details. RESULTS In the present study, a thermophilic recombinant esterase was immobilized by entrapment in chitosan/calcium/alginate-blended beads for the first time. The 0.5 mg mL(-1) purified recombinant esterase was entrapped in 1% chitosan, 2% alginate, and 0.7 M CaCl2 blended beads. The results showed that immobilization yield and entrapment efficiency of the entrapped esterase were 69.5% and 80.4%, respectively. SEM micrograph showed that the surface of the beads resembled a mesh and very compact structures. Chitosan/calcium/alginate-blended beads exhibited an 18.8% swelling ratio and had a moderate porous structure. The entrapment technique highly enhanced the thermostability of the esterase and shifted its optimum temperature from 65 to 80 degrees C. The immobilized esterase was very stable in a wide range of pH (8.5-11) displaying maximum activity at pH 9. ZnCl2 slightly increased the activity of immobilized esterase whereas several metal ions reduced the enzyme activity. When the enzyme was immobilized in chitosan/calcium/alginate-blended beads, its K-m increased about 2 times and V-max value decreased almost 1.5 times. Immobilization allowed repeated uses of the esterase having good operational stability in a continuous process. CONCLUSION The results revealed that the immobilization of a thermophilic recombinant esterase by entrapment in chitosan/calcium/alginate-blended beads exhibited considerably better compared to other immobilization processes with various entrapment strategies. (c) 2021 Society of Chemical Industry (SCI).Book Part Citation - Scopus: 6Recent Advances in Chitosan-Based Systems for Delivery of Anticancer Drugs(Springer, 2020) Polat, Mehmet; Polat, HürriyetProblems in transporting drug molecules to tumor sites in required dose or constitution lead to low efficacy and significant side effects. Shielding the drug molecules in micelles, liposomes, or nanoparticles is a major line of investigation to improve chemotherapeutic treatment. Though compatibility for proper envelopment of the drug and timely release at the tumor site are required of such a carrier, protecting its own physicochemical and morphological integrity during transport is another precondition. Because of its superior polymerization capability, biocompatibility, pH dependence, and charging characteristics, chitosan has been in the forefront of potential drug carriers. Numerous synthesis routes for chitosan-based nanocarriers have been suggested to the extent that a search of the literature published since 2000 with the keywords “novel + nano + chitosan” in the title results in 527 articles, indicating the bewildering quality and quantity of the new information. This review was carried out not only to peruse this large amount of work on chitosan-based anticancer drug delivery but also to extract manageable patterns from numerous synthesis routes. The main conclusion is that the synthesis methods suggested in literature can be combined into two main routes, and the degree of hydrophobicity of the drug determines which route should be followed. © Springer Nature Singapore Pte Ltd. 2019.Article Citation - WoS: 16Citation - Scopus: 19Designing of Spherical Chitosan Nano-Shells With Micellar Cores for Solvation and Safeguarded Delivery of Strongly Lipophilic Drugs(Elsevier Ltd., 2017) Cihan, Esra; Polat, Mehmet; Polat, HürriyetChitosan is a very effective biopolymer for drug delivery purposes due to its biocompatibility, positive charge and exceptionally pH sensitive degradability behavior in an aqueous medium. Nevertheless, its inability for dissolving lipophilic drug active material and the difficulties in controlling the size and shape of the synthesized particles in nanometer range are critical drawbacks in its effective use. In this study, a synthesis procedure which addresses both issues simultaneously is presented. The procedure is based on initial dissolution of lipophilic drug molecules within the hydrophobic cores of the micelles of a bio-compatible block-copolymer by ionic gelation and subsequent formation of a chitosan shell by polymerization around the micellar structures. Well-formed, hollow and perfectly spherical chitosan particles (nano-shells) in the 30–300 nm size range could be successfully manufactured. Characterization by STEM, TEM, AFM, FTIR and DLS, DLS-LDV techniques showed clearly that the drug was successfully incorporated into the chitosan structure. It was demonstrated that the particles enveloped the micelle(s) of a Pluronic copolymer (P-123) whose hydrophobic cores contained a strongly hydrophobic drug Probucol. The chitosan nano-shells are expected to act as an agent protecting the integrity of the drug-loaded micelles in the body fluid while providing a pH sensitive release medium. The drug uptake by the chitosan particles was very high. A very sharp increase in the amount of the drug released with a slight change in the acidity of the medium was an indication of the potential of the manufactured chitosan nano-shells as pH sensitive, target specific delivery vehicles for drug release.Article Citation - WoS: 15Citation - Scopus: 15Bodipy-Conjugated Chitosan Nanoparticles as a Fluorescent Probe(Taylor and Francis Ltd., 2017) Bor, Gizem; Üçüncü, Muhammed; Emrullahoğlu, Mustafa; Tomak, Aysel; Şanlı Mohamed, GülşahRecently, development of fluorescent nanoparticle-based probes for various bioimaging applications has attracted great attention. This work aims to develop a new type fluorescent nanoparticle conjugate and evaluate its cytotoxic effects on A549 and BEAS 2B cell lines. Throughout the study, ionically crosslinked chitosan nanoparticles (CNs) were conjugated with carboxylated 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY-COOH). The results of conjugates (BODIPY-CNs) were investigated with regard to their physic-chemical, optical, cytotoxic properties and cellular internalization. The morphology of BODIPY-CNs was found to be spherical in shape and quite uniform having average diameter of 70.25 ± 11.99 nm. Cytotoxicty studies indicated that although BODIPY-COOH itself was quite toxic on both A549- and BEAS 2B-treated cells, CNs increased the cell viability of both cell lines via conjugation to BODIPY-COOH fluorescent molecule up to 67% for A549 and 74% for BEAS 2B cells. These results may suggest a possible utilization of the new fluorescent nanoparticle-based probe for bioimaging in biology and medicine.Article Citation - WoS: 16Citation - Scopus: 18Cytotoxic and Cytostatic Side Effects of Chitosan Nanoparticles as a Non-Viral Gene Carrier(Elsevier Ltd., 2016) Bor, Gizem; Mytych, Jennifer; Zebrowski, Jacek; Wnuk, Maciej; Şanlı Mohamed, GülşahAlthough chitosan nanoparticles (CNs) became a promising tool for several biological and medical applications owing to their inherent biocompatibility and biodegradability features, studies regarding their effects on cytotoxic and cytostatic properties still remain insufficient. Therefore, in the present study, we decided to perform comprehensive analysis of the interactions between CNs–pKindling-Red-Mito (pDNA) and different cell line models derived from blood system and human solid tissues cancers. The resulting CNs-pDNA was investigated in terms of their cellular uptake, transfection efficiency, and physico-chemical, cytotoxic and cytostatic properties. The nanoparticles showed high encapsulation efficiency and physical stability for various formulations even after two days time period. Moreover, high gene expression levels were observed after 96 h of transfection. CNs-pDNA treatment, despite the absence of oxidative stress induction, caused cell cycle arrest in G0/G1 phase and as a consequence led to premature senescence which turned out to be both p21-dependent and p21-independent. Also, observed DNMT2 upregulation may suggest the activation of different pathways protecting from the results of CNs-mediated stress. In conclusion, treatment of different cell lines with CNs-pDNA showed that their biocompatibility was limited and the effects were cell type-dependent.Article Citation - WoS: 140Citation - Scopus: 171Physicochemical Characterization of Chitosan Extracted From Metapenaeus Stebbingi Shells(Elsevier Ltd., 2011) Küçükgülmez, Aygül; Çelik, Mehmet; Yanar, Yasemen; Şen, Didem; Polat, Hürriyet; Kadak, Ali EslemIn this study, chitosan was extracted from Metapenaeus stebbingi shells. In order to determine physicochemical characteristics of the extracted chitosan, the yield, moisture and ash contents, degree of deacetylation, molecular weight, water and fat binding capacities, apparent viscosity and colour properties were measured using a variety of techniques including Fourier transform infrared spectroscopy, scanning electron microscopy and X-ray diffraction. In addition, the physicochemical characteristics of the chitosan extracted from M. stebbingi shells were compared to commercial chitosan. The degree of deacetylation was calculated by the titration method and elemental analysis. The molecular weight was determined by viscosimetric methods. The results of the study indicate that shrimp shells are a rich source of chitosan as 17.48% of the shell's dry weight is consisted of this material. Extracted chitosan exhibited a lower molecular weight, higher degree of deacetylation, higher viscosity and higher water and fat binding capacities compared to the commercial chitosan.Article Citation - WoS: 15Citation - Scopus: 21Preparation, Characterization and Optimization of Chitosan Nanoparticles as Carrier for Immobilization of Thermophilic Recombinant Esterase(Taylor and Francis Ltd., 2011) İlgü, Hüseyin; Turan, Taylan; Şanlı Mohamed, GülşahImmobilization of biologically important molecules on myriad nano-sized materials has attracted great attention. Through this study, thermophilic esterase enzyme was obtained using recombinant DNA technology and purified applying one-step His-Select HF nickel affinity gel. The synthesis of chitosan was achieved from chitin by deacetylation process and degree of deacetylation was calculated as 89% by elemental analysis. Chitosan nanoparticles were prepared based on the ionic gelation of chitosan with tripolyphosphate anions. The physicochemical properties of the chitosan and chitosan nanoparticles were determined by several methods including SEM (Scanning Electron Microscopy), FT-IR (Fourier Transform Infrared Spectroscopy) and DLS (Dynamic Light Scattering). The morphology of chitosan nanoparticles was spherical and the nanospheres' average diameter was 75.3 nm. The purified recombinant esterase was immobilized efficiently by physical adsorption onto chitosan nanoparticles and effects of various immobilization conditions were investigated in details to develope highly cost-effective esterase as a biocatalyst to be utilized in biotechnological purposes. The optimal conditions of immobilization were determined as follows; 1.0 mg/mL of recombinant esterase was immobilized on 1.5 mg chitosan nanoparticles for 30 min at 60C, pH 7.0 under 100 rpm stirring speed. Under optimized conditions, immobilized recombinant esterase activity yield was 88.5%. The physicochemical characterization of enzyme immobilized chitosan nanoparticles was analyzed by SEM, FT-IR and AFM (Atomic Force Microscopy).Article Citation - WoS: 17Citation - Scopus: 21Humidity Sensing Properties of Chitosan by Using Quartz Crystal Microbalance Method(American Scientific Publishers, 2012) Havare, A. Kemal; İlgü, Hüseyin; Okur, Salih; Şanlı Mohamed, GülşahHumidity adsorption kinetics of chitosan films was investigated by quartz crystal microbalance (QCM) technique. In this study, chitosan was synthesized from chitin by deacetylation process in 2.0% (v/v) acetic acid solution and then coated on QCM to measure humidity response. The Langmuir model was used to determine the adsorption rates and Gibbs free energy for various relative humidity between 11% and 94%. The average Gibbs free energy for adsorption was obtained as 12.93 kJ/mol. Our reproducible experimental results show that chitosan films are very sensitive to relative humidity changes at room temperature. Copyright © 2012 American Scientific Publishers All rights reserved.Article Citation - WoS: 20Citation - Scopus: 23Rhodamine-Immobilised Electrospun Chitosan Nanofibrous Material as a Fluorescence Turn-On Hg2+ Sensor(John Wiley and Sons Inc., 2016) Horzum, Nesrin; Mete, Derya; Karakuş, Erman; Üçüncü, Muhammed; Emrullahoğlu, Mustafa; Demir, Mustafa MuammerA turn-on fluorescence sensing system for mercuric (Hg2+) ions relying on a modified rhodamine B–chitosan fluorophore moiety was developed. This novel sensing approach relies on the simultaneous electrospinning of chitosan and rhodamine B hydrazide with phenylisothiocyanate functionality in hexafluoroisopropanol solution at 3.4 kV cm−1. The electrospun mats exhibited not only considerably enhanced fluorescence intensity in the presence of mercury ions, a result attributed to the ring opening of the spirolactam unit of the rhodamine-based fluorophore, but also a remarkably high sensitivity and selectivity toward Hg2+. In effect, the strategy has the potential to open new avenues in the design and development of other high-performance nanofibrous sensing materials for detecting target metal species of environmental interest.Article Citation - WoS: 14Citation - Scopus: 15Chitosan-Immobilized Pumice for the Removal of As(v) From Waters(Springer Verlag, 2014) Turan, Dilek; Kocahakimoğlu, Cemre; Boyacı, Ezel; Sofuoğlu, Sait Cemil; Eroğlu, Ahmet EminA novel sorbent, chitosan-immobilized pumice, has been prepared for the sorption of As(V) from waters prior to its determination by hydride generation atomic absorption spectrometry. The success of the immobilization has been checked with such characterization techniques as scanning electron microscopy, thermal gravimetric analysis, and elemental analysis. Points of zero charge of the sorbents were determined with potentiometric mass titration. Batch-type equilibration studies have shown that the novel sorbent can be employed at a wide pH range resulting in quantitative sorption (>90 %) at pH 3.0-7.0 and greater than 70 % sorption at pH >8.0. These results demonstrate the advantage of immobilizing chitosan onto pumice, because, under the same conditions, pumice displays <20 % sorption toward As(V), whereas chitosan gives approximately 90%sorption only at pH 3.0. The validity of the method was verified through the analysis of ultrapure, bottled drinking, and tap water samples spiked with arsenate; the respective sorption percentages of 93.2 (±0.7), 89.0 (±1.0), and 80.9 (±1.3) were obtained by batch-type equilibration. Arsenic sorption was also examined in the presence of common interfering ions resulting in competing effects of PO3- 4 and NO- 3on As(V) adsorption.