Chemistry / Kimya

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  • Article
    Citation - WoS: 7
    Citation - Scopus: 8
    Thermoalkalophilic Recombinant Esterase Entrapment in Chitosan/Calcium Beads and Its Characterization
    (Wiley, 2021) Tercan, Cisem; Sürmeli, Yusuf; Şanlı Mohamed, Gülşah
    BACKGROUND 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).
  • Article
    Citation - WoS: 16
    Citation - Scopus: 19
    Designing 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, Hürriyet
    Chitosan 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: 15
    Citation - Scopus: 15
    Bodipy-Conjugated Chitosan Nanoparticles as a Fluorescent Probe
    (Taylor and Francis Ltd., 2017) Bor, Gizem; Üçüncü, Muhammed; Emrullahoğlu, Mustafa; Tomak, Aysel; Şanlı Mohamed, Gülşah
    Recently, 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: 15
    Citation - Scopus: 21
    Preparation, 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, Gülşah
    Immobilization 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: 70
    Citation - Scopus: 82
    Sorption Efficiency of Chitosan Nanofibers Toward Metal Ions at Low Concentrations
    (American Chemical Society, 2010) Horzum Polat, Nesrin; Boyacı, Ezel; Eroğlu, Ahmet Emin; Shahwan, Talal; Demir, Mustafa Muammer
    Chitosan fibers showing narrow diameter distribution with a mean of 42 nm were produced by electrospinning and utilized for the sorption of Fe(III), Cu(II), Ag(I), and Cd(II) ions from aqueous solutions. The ion concentrations in the supernatant solutions were determined using inductively coupled plasma-mass spectrometry (ICP-MS). The filtration efficiency of the fibers toward these ions was studied by both batch and microcolumn methods. High efficiency in sorption of the metal ions was obtained in the both methods. The effects of sorbent amount (0.10−0.50 mg), shaking time (15−120 min), initial metal ion concentration (10.0−1000.0 μg·L−1), and temperature (25 and 50 °C) on the extent of sorption were examined. The sorbent amount did not significantly alter the efficiency of sorption; however, shaking time, temperature, and metal ion concentration were found to have a strong influence on sorption. By virtue of its mechanical integrity, the applicability of the chitosan mat in solid phase extraction under continuous flow looks promising.