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Adsorption behaviours of lysozyme onto poly-hydroxyethyl methacrylate cryogels containing methacryloyl antipyrine-Ce(III)
(Taylor & Francis As, 2018) Baysal, Zubeyde; Aksoy, Eyyup; Dolak, Ibrahim; Ersoz, Arzu; Say, Ridvan
In this study, Ce3+-based cryogel with methacryloyl antipyrine (MAAP) and 2-hydroxyethyl methacrylic acid (HEMA) [p(HEMA-MAAP-Ce3+] was prepared. MAAP-Ce3+ complex was characterized by UV-near infrared and energy-dispersive X-ray spectroscopy. Cryogel was characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and swelling test. Pore size of the cryogel was found to be about 30-50 mu m. The effects of flow rate, pH, temperature, and initial enzyme concentration have been investigated. Maximum adsorption capacity was found to be 57.84 mg g(-1) cryogel at pH 6.0. After seven times of adsorption-desorption cycles of same cryogel, it was observed that there is negligible decrease in the adsorption capacity. [GRAPHICS] .
Adsorption of cellulase on poly(2-hydroxyethyl methacrylate) cryogels containing phenylalanine
(Tubitak Scientific & Technological Research Council Turkey, 2016) Yavuz, Murat; Cakir, Oguz; Baysal, Zubeyde
The aim of this study was to prepare a supermacroporous cryogel that can be used for the adsorption of cellulase. The macroporous cryogel of poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-phenylalanine) [p(HEMA-MAPA)] was prepared by copolymerization of 2-hydroxyethyl methacrylate (HEMA) with a functional monomer of N-methacryloyl-L-phenylalanine (MAPA). The cryogel was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, and swelling tests. The effects of several parameters such as medium pH, temperature, ionic strength, and flow rate on cellulase adsorption were also investigated. Maximum cellulase adsorption was observed at 25 degrees C and pH 4.0. Furthermore, adsorbed cellulase was desorbed from the cryogel by using 1.0 M NaCl. The p(HEMA-MAPA) cryogel could be used many times without the cellulase adsorption capacity decreasing significantly.
Adsorption of Ni(II) from aqueous solution by Bacillus subtilis
(Taylor & Francis Inc, 2012) Bulut, Yasemin; Gul, Ayten; Baysal, Zubeyde; Alkan, Huseyin
This work reports the application of Bacillus subtilis as adsorbent for the removal of Ni(II) from aqueous solution. Batch experiments were conducted to study the effects of several parameters such as, contact time, initial concentration of adsorbate (25-200 mg L-1), temperature (298-318 K), and adsorbent dose (0.05-0.4 g) on Ni(II) adsorption. Equilibrium adsorption isotherms and kinetics were also investigated. The equilibrium experimental data were analyzed by Freundlich and Langmuir models. The kinetic data obtained with different initial concentration and temperature were analyzed using a pseudo-first-order, pseudo-secondorder, and intraparticle diffusion equations. The results showed that this novel adsorbent had a high adsorption capacity, making it suitable for use in the treatment of Ni(II)-enriched wastewater.
Development of molecular imprinting-based smart cryogels for selective recognition and separation of serum cytochrome-c as a biochemical indicator
(Elsevier Sci Ltd, 2021) Canpolat, Gurbet; Dolak, Ibrahim; Onat, Ruken; Kecili, Rustem; Baysal, Zubeyde; Ziyadanogullari, Berrin; Ersoz, Arzu
The recognition and detection of proteins has an important role in the fields of biochemistry and medicine in term of diagnosis and prognosis. In this study, molecularly imprinted cryogel (MIP) was synthesized for the selective recognition of Cytochrome-c (Cyt-c) by cryopolymerization techniques with the lanthanide-chelate approach using (MAAP)2-Ce(III) as complex functional monomer and Cyt-c as a template protein. The incorporation of template with monomer was characterized by near- infrared (Near-IR) spectroscopy. The binding capacity was optimized in accordance with numerous experimental parameters including pH, initial Cyt-c concentration, flow rate, temperature and ionic strength. As a result, the maximum binding capacity reached to 98.33 mg g-1 in a buffer system with a pH of 6.0. Beside of the excellent reusability performance of prepared MIP, it was successfully applied to separation of template Cyt-c in the existence of hemoglobin (Hb) and myoglobin (Mb) chosen as interfering proteins. The relative selectivity constants of the Cyt-imprinted polymer were determined as 13.84 for the Cyt/Mb pair and 16.05 for the Cyt/Hb pair. Consequently, a high selectivity, simple and cheap strategy for efficient separation of proteins was provided with this novel MIP cryogel.
A novel lanthanide-chelate based molecularly imprinted cryogel for purification of hemoglobin from blood serum: An alternative method for thalassemia diagnosis
(Elsevier Sci Ltd, 2020) Dolak, Ibrahim; Canpolat, Gurbet; Onat, Ruken; Kecili, Rustem; Baysal, Zubeyde; Ziyadanogullari, Berrin; Ersoz, Arzu
Purification and analyzing of proteins is an essential means for understanding their function and diseases associated with their lack or defect. In this research, a new lanthanide-chelate based molecularly imprinted polymer (MIP) was synthesized for selective separation of Hemoglobin (Hb) from human serum in the presence of various interference molecules. The Hb-imprinted polymer was prepared by using complex functional monomer N-methacryloylamido antipyrine (MAAP)-Ce(III) and 2-Hydroxyethyl methacrylate (HEMA) in accordance of cryopolymerization techniques. The nonimprinted cryogel (NIP) was also prepared at same polymerization conditions in the absence of template Hb molecule. The effects of pH, initial Hb concentration, flow rate, temperature and ionic strength on the binding capacity of both imprinted and nonimprinted cryogels was investigated. The maximum binding capacity for the MIP column was found to be as 79.41 mg g(-1) dry cryogel, that is four times higher than the NIP column under the optimum conditions (pH 5.0, flow rate: 1.0 mL min(-1), T: 25 degrees C). Moreover, selectivity experiments were performed by using two interference proteins as myoglobin (Mb) and cytochrome c (Cyt-c) and the relative selectivity coefficients (k') for Hb/Mb and Hb/Cyt-c pairs were determined as 36.59 and 37.22, respectively.
Novel QCM and SPR sensors based on molecular imprinting for highly sensitive and selective detection of 2,4-dichlorophenoxyacetic acid in apple samples
(Elsevier, 2019) Cakir, Oguz; Bakhshpour, Monireh; Yilmaz, Fatma; Baysal, Zubeyde
This study aims to develop molecularly imprinted based quartz crystal microbalance (QCM) and surface plasmon resonance (SPR) sensors for highly sensitive and selective detection of 2,4-dichlorophenoxyacetic acid (2,4-D) and to determine their accuracy and precision by liquid chromatography-tandem mass spectrometry (LC-MS/MS) as a reference technique. Here, we synthesized non-imprinted (NIP) and 2,4-D-imprinted (MIP) [ethylene glycol dimetacrylate-N-metacryloyl-(L)-tryptophan methyl ester-p(EGDMA-MATrp)] polymeric nanofilms by using molecular imprinting technique. MIP and NIP nanofilms were characterized by fourier transform infrared spectroscopy attenuated total reflectance (FTIR-ATR), atomic force microscope (AFM), contact angle and ellipsometer measurements. The molecular imprinting procedures were successfully carried out and it was found that the prepared polymeric surfaces were highly desirable for sensitive recognition by QCM and SPR sensors. Competitive experiments for the sensors revealed that MW nanofilms were found to show more sensitivity and selectivity than NIP ones. The sensor responses have a good linear relationship with 2,4-D concentrations in the range of 0.23-8.0 nM with a limit of detection at 20.17 ng/L for QCM and 24.57 ng/L for SPR sensors. In conclusion, both QCM and SPR sensor systems showed good accuracy and precision, with recovery percentages between 90 and 92% and 87-93%, respectively. Furthermore, they have a fast response time, reusability, high selectivity and sensitivity and low limit of detection.
Optimization and enhanced production of ?-amylase and protease by a newly isolated Bacillus licheniformis ZB-05 under solid-state fermentation
(Springer, 2013) Karatas, Hakan; Uyar, Fikret; Tolan, Veysel; Baysal, Zubeyde
Eight different agro-residues were tested for alpha-amylase and protease production by using Bacillus licheniformis ZB-05. Among them, rice husk (RH) was proved as the best substrate for two enzymes (alpha-amylase 443 U/g and protease 469,000 U/g). Maximum enzyme production was observed to be 30 % initial moisture, with a growth period of 36 h in 20 and 30 % inoculum volumes for alpha-amylase and protease, respectively. The best enzyme recovery from solid mass was obtained when extracted with tap water. Among the tested various nitrogen sources, 1 % ammonium sulphate followed by 2 % Bacto liver, 2 % ammonium sulphate and 1 % Bacto casaminoacid served as the best inorganic and organic nitrogen sources for alpha-amylase and protease production, respectively. As additional carbon sources, 2 % soluble starch enhanced alpha-amylase production, while 1 % maltose enhanced protease production.
OPTIMIZATION OF ?-AMYLASE PRODUCTION BY Bacillus subtilis RSKK96: USING THE TAGUCHI EXPERIMENTAL DESIGN APPROACH
(Taylor & Francis Inc, 2011) Uysal, Ersin; Akcan, Nurullah; Baysal, Zubeyde; Uyar, Fikret
In this study, the Taguchi experimental design was applied to optimize the conditions for alpha-amylase production by Bacillus subtilis RSKK96, which was purchased from Refik Saydam Hifzissihha Industry (RSHM). Four factors, namely, carbon source, nitrogen source, amino acid, and fermentation time, each at four levels, were selected, and an orthogonal array layout of L-16(4(5)) was performed. The model equation obtained was validated experimentally at maximum casein (1%), corn meal (1%), and glutamic acid (0.01%) concentrations with incubation time to 72 h in the presence of 1% inoculum density. Point prediction of the design showed that maximum alpha-amylase production of 503.26 U/ mg was achieved under optimal experimental conditions.
p(HEMA)-Pd(II) and p(HEMA-MAH)-Pd(II) Microspheres: Efficient, Recyclable and Ligand-Free Catalyst for Suzuki-Miyaura Cross-Coupling Reaction in Water
(Taylor & Francis Inc, 2016) Durap, Feyyaz; Baysal, Akin; Elma, Duygu; Aydemir, Murat; Ok, Ozge; Baysal, Zubeyde
In this study, the authors report the synthesis and characterization of new poly(hydroxyethyl methacrylate) palladium(II) (p (HEMA)-Pd(II)) and poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) palladium(II), p(HEMA-MAH)Pd(II) microsphere catalysts, and their catalysis in Suzuki-Miyaura coupling reactions of aryl bromides with phenylboronic acid in water. The microsphere catalysts were characterized by SEM, ICP-MS, FTIR, and diffuse-reflactance UV-visible spectroscopy. The p(HEMA)-Pd(II) and p(HEMA-MAH)-Pd(II) microsphere catalysts were active catalysts in Suzuki-Miyaura coupling reactions of arylbromides with phenylboronic acid affording biphenyls in high yield. Recycling experiments showed that the p(HEMA)-Pd(II) and p(HEMA-MAH)-Pd(II) microsphere catalysts could be used seven or ten times with essentially no loss in activity in the Suzuki-Miyaura coupling reactions.
Pesticide analysis with molecularly imprinted nanofilms using surface plasmon resonance sensor and LC-MS/MS: Comparative study for environmental water samples
(Elsevier Science Sa, 2019) Cakir, Oguz; Baysal, Zubeyde
SPR sensor chip nanofilms were prepared using a molecular imprinting method. Characterization of nanofilms was achieved by fourier-transform infrared spectroscopy attenuated total reflectance (FTIR-ATR), contact angle, atomic force microscope, and ellipsometer. Kinetic and affinity binding of pesticides were investigated by binding pesticide imprinted and nonimprinted sensor chips to the SPR sensor. Comparative adsorption experiments for SPR sensors were performed to display the selectivity of pesticide imprinted nanofilms. Analysis of the SPR sensors revealed that imprinted nanofilms showed more sensitivity and selectivity than the nonimprinted ones for pesticide determination. Exact masses of molecular and fragmentation ions of pesticides were determined by liquid chromatography ion trap time-of-flight mass spectroscopy (LC/MS-IT-TOF). According to these exact masses and fragmentation ions, an LC-MS/MS method was prepared and validated for qualitative and quantitative analysis of the studied pesticides. The limit of detection values of dimethoate and carbofuran were found to be 16.92 ng L-1 and 20.47 ng L-1 in the LC-MS/MS method and 8.37 ng L-1 and 7.11 ng L-1 in the SPR sensor system, respectively. The accuracy and precision of both methods were determined by comparison of six replicates at three concentrations (50, 250, and 1000 ng L-1), using the same environmental water matrix for each pesticide. SPR sensors showed good accuracy, with recovery percentages between 90 and 95 for both pesticides. The results showed that SPR sensors have higher selectivity and sensitivity, and lower detection limits compared to LC-MS/MS.
Preconcentration with Bacillus subtilis-Immobilized Amberlite XAD-16: Determination of Cu2+ and Ni2+ in River, Soil, and Vegetable Samples
(Taylor & Francis Inc, 2015) Okumus, Veysi; Ozdemir, Sadin; Kilinc, Ersin; Dundar, Abdurrahman; Yuksel, Uyan; Baysal, Zubeyde
Solid-phase extraction (SPE) method was developed for the preconcentration of Cu2+ and Ni2+ before their determination by inductively coupled plasma optical emission spectrometry (ICP-OES). Bacillus subtilis-immobilized Amberlite XAD-16 was used as biosorbent. Effects of critical parameters such as pH, flow rate of samples, amount of Amberlite XAD-16 and biosorbent, sample volume, eluent type, and volume and concentration of eluent on column preconcentration of Cu2+ and Ni2+ were optimized. Applicability of the method was validated through the analysis of the certified reference tea sample (NCS ZC73014). Sensitivity of ICP-OES was improved by 36.4-fold for Cu2+ and 38.0-fold for Ni2+ by SPE-ICP-OES method. Limit of quantitation (LOQ) was found to be 0.7 and 1.1ng/ml for Cu2+ and Ni2+, respectively. Concentrations of Cu2+ and Ni2+ were determined by ICP-OES after application of developed method. Relative standard deviations (RSDs) were lower than 4.9% for Cu2+ and 7.9% for Ni2+. The Tigris River that irrigates a large agricultural part of Southeast Turkey is polluted by domestic and industrial wastes. Concentrations of Cu2+ and Ni2+ were determined in water, soil, and some edible vegetables as a biomonitor for heavy metal pollution.
Preparation and Use of Poly(hydroxyethyl methacrylate) Cryogels Containing L-Histidine for ?-Casein Adsorption
(Wiley-Blackwell, 2013) Yavuz, Murat; Baysal, Zubeyde
The objective of this study was to determine -casein adsorption by using supermacroporous poly(2-hydroxyethyl methacrylate-N-methacryloyl-(l)-histidine methyl ester) [p(HEMA-MAH)] cryogel. -Casein adsorption properties of p(HEMA-MAH) cryogel were studied for the application of -casein purification. The cryogel was produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine and ammonium persulfate pairs in an ice bath. P(HEMA-MAH) cryogel was characterized by swelling tests, Fourier transform infrared spectroscopy, and scanning electron microscopy. The effects of the flow rate, pH, temperature, initial -casein concentration, and ionic strength on the adsorption efficiency of cryogel were studied. The equilibrium swelling degree of the p(HEMA-MAH) cryogel was 6.73g H2O/g cryogel. -Casein adsorption capacity of p(HEMA-MAH) cryogel from aqueous solution was estimated as 31.17mg/g cryogel. It was also observed that -casein could be repeatedly adsorbed and desorbed with p(HEMA-MAH) cryogel without significant loss in the adsorption capacity.
Preparation of a new quartz crystal microbalance sensor based on molecularly imprinted nanofilms for amitrole detection
(Biointerface Research Applied Chemistry, 2018) Cakir, Oguz; Yilmaz, Fatma; Baysal, Zubeyde; Denizli, Adil
Quartz crystal microbalance (QCM) sensors have been used to detect a variety of biomolecules due to their simplicity, specificity and sensitivity, real-time measurement, low cost and no labeling requirements. A new QCM sensor was prepared by using molecular imprinting method for selective recognition of amitrole. N-metacryloyl-(L)-tryptophan methyl ester (MATrp) was selected as a proper functional monomer and polymerized with ethylene glycol dimethacrylate (EGDMA). Pesticide imprinted poly(ethylene glycol dimetacrylate-N-metacryloyl-(L)-tryptophan methyl ester) [poly(EGDMA-MATrp)] nanofilms were attached to gold surfaces of QCM sensor chips and were characterized by several techniques such as atomic force microscope (AFM), an ellipsometer, FTIR-ATR and contact angle measurements. Kinetic and affinity binding of amitrole was investigated by binding the pesticide imprinted and nonimprinted sensor chips to QCM sensor chips. The imprinted nanofilms were found to show more sensitivity towards the target molecule than the nonimprinted ones. Furthermore, adsorption kinetics were determined by passing pesticide solutions at different concentrations through QCM sensor systems. The most proper model was found to be Langmuir adsorption model for these affinity systems. In addition, competitive adsorption experiments were performed to display selectivity of the pesticide imprinted nanofilms. The prepared sensor was also efficiently applied for the selective detection of amitrole in green pepper.
Preparation of poly(hydroxyethyl methacrylate) cryogels containing L-histidine for insulin recognition
(Elsevier, 2013) Cavus, Aynur; Baysal, Zubeyde; Alkan, Huseyin
In the present study, affinity adsorption technique was studied for insulin adsorption. Firstly, insulin-imprinted supermacroporous cryogel was prepared for the insulin adsorption. N-methacryloyl-(L)-histidine methyl ester (MAH) was chosen as the monomer. Insulin was complexed with MAH, and insulin-imprinted p(HEMA MAH) [insulin-(MIP)] cryogel was prepared by free radical polymerization with 2-hydroxyethyl methacrylate (HEMA), N,N,N',N'-tetramethylethylenediamine (TEMED) and ammonium persulfate (APS) in an ice bath. Then, insulin was removed from the cryogel by using 0.1 M glycine-HCl buffer (pH: 3.5). The characterization of the cryogel was carried out by using scanning electron microscopy (SEM) and swelling test. The equilibrium swelling ratios of the cryogels were found to be 8.56 +/- 0.42 g H2O/g polymer for p(HEMA) and 7.20 +/- 0.36 g H2O/g polymer for insulin-p(HEMA MAH). Insulin adsorption experiments were performed under different conditions, such as flow rate, medium pH, initial insulin concentration and ionic strength. It was observed that insulin could be repeatedly adsorbed and desorbed with MIP cryogel without any significant decrease in the adsorption capacity. (C) 2013 Elsevier B.V. All rights reserved.
PRODUCTION AND CHARACTERIZATION OF PARTIALLY PURIFIED EXTRACELLULAR THERMOSTABLE ?-AMYLASE BY Bacillus subtilis IN SUBMERGED FERMENTATION (SmF)
(Taylor & Francis Inc, 2011) Ozdemir, Sadin; Matpan, Fatma; Guven, Kemal; Baysal, Zubeyde
A Bacillus strain was isolated from soil samples from the campus area of Dicle University. Based on 16S ribosomal RNA sequencing, the microorganism was closely related to Bacillus subtilis. Effects of different culture medium, incubation time, carbon and nitrogen sources, and various starches, flours, and chemicals on alpha-amylase production were examined. Maximum enzyme production (7516 U/mL) was obtained in a basal medium A containing 0.05% Tween 40 in 24 h. Partially purified enzyme showed maximum activity at 60 degrees C with an optimum pH of 6.0. The effects of 0.2% detergents (sodium dodecyl sulfate [SDS], CHAPS [3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate], and commercial detergent Omo Matic) on partially purified enzyme activity over a period of time (15-150 min) were examined and the order of inhibition effect from the most to the least was found as SDS> Omo Matic> CHAPS. Different metal ions inhibited alpha-amylase activity at low concentrations (1.5 mM). Co2+ was a mild inhibitor and Hg2+ and Cd2+ were potent inhibitors, whereas Ca2+ and Mg2+ increased the enzyme activity. At 20 mM, Ca2+ enhanced enzyme activity, and different Ca2+ concentrations (10-300 mM) were studied.
Production of lipase by a newly isolated Bacillus coagulans under solid-state fermentation using melon wastes
(Humana Press Inc, 2007) Alkan, Huseyin; Baysal, Zubeyde; Uyar, Fikret; Dogru, Mehmet
An extracellular lipase was produced by Bacillus coagulans by solid-state fermentation. Solid waste from melon was used as the basic nutrient source and was supplemented with olive oil. The highest lipase production (78,069 U/g) was achieved after 24 h of cultivation with 1% olive oil enrichment. Enzyme had an optimal activity at 37 degrees C and pH 7.0, and sodium dodecyl sulfate increased lipase activity. NH4NO3 increased enzyme production, whereas organic nitrogen had no effect. The effect of the type of carbon sources on lipolytic enzyme production was also studied. The best results were obtained with starch and maltose (148,932 and 141,629 U/g, respectively), whereas a rather low enzyme activity was found in cultures grown on glucose and galactose (approx 118,769 and 123,622 U/g, respectively). Enzyme was inhibited with Mn+2 and Ni+2 by 68 and 74%, respectively. By contrast, Ca+2 enhanced enzyme production by 5%.
Removal of methylene blue from aqueous solutions onto Bacillus subtilis: determination of kinetic and equilibrium parameters
(Desalination Publ, 2013) Ayla, Ayseguel; Cavus, Aynur; Bulut, Yasemin; Baysal, Zubeyde; Aytekin, Cetin
In the present study, Bacillus subtilis, a Gram-positive bacteria, was used in dried biomass form as biosorbent for removal of methylene blue (MB) from aqueous solutions. Batch adsorption tests were performed at different contact times, temperatures, pH, adsorbent doses, and initial dye concentration. The adsorption isotherms are described by means of the Langmuir and Freundlich isotherms. It was found that the Langmuir equation fit better than the Freundlich equation. Maximum biosorption capacity was found to be 169.49, 178.57, and 181.82mgg(-1) at 298, 308, and 318K, respectively. The adsorption kinetics of MB could be described by the pseudo-second-order reaction model. The activation energy of the biosorption (E-a) was determined as 11.30kJmol(-1) at initial concentration of 50mgL(-1). Free energy of adsorption (G degrees), enthalpy (H degrees), and entropy (S degrees) changes were calculated to predict the nature of adsorption. The estimated values for G degrees were -21.22, -22.02, and -23.01kJmol(-1) at 298, 308, and 318K, respectively. The enthalpy changes and entropy of adsorption were 5.47kJmol(-1) and 89.43Jmol(-1), respectively. The experimental data obtained in the present study indicate that B. subtilis is a suitable and inexpensive adsorbent which can be used for dye removal in wastewater treatment processes.
Selective removal of the autoantibodies from rheumatoid arthritis patient plasma using protein A carrying affinity cryogels
(Elsevier, 2010) Alkan, Huseyin; Bereli, Nilay; Baysal, Zubeyde; Denizli, Adil
Rheumatoid arthritis is a chronic, progressive, deabilitating autoimmune disease that occurs in approximately 1% of adults. Rheumatoid arthritis is characterized by chronic polyarthritis and destruction of multiple joints. In this study, IgM-antibody removal from human plasma with supermacroporous poly(hydroxyethyl methacrylate) [PHEMA] cryogel carrying protein A has been evaluated. The PHEMA cryogel was prepared by bulk polymerization which proceeds in an aqueous solution of monomer frozen inside a plastic syringe (cryo-polymerization). After thawing, the PHEMA cryogel contains a continuous matrix having interconnected macropores of 10-200 mu m size. Pore volume in the PHEMA cryogel was 71.6%. Protein A molecules were covalently immobilized onto the PHEMA cryogel via cyanogen bromide (CNBr) activation. The PHEMA cryogel was contacted with blood in in vitro system for the determination of blood-compatibility. The supermacroporous structure of the PHEMA cryogel makes it possible to process blood cells without blocking the cryogel column. IgM-antibody adsorption capacity decreased significantly with the increase of the plasma flow-rate. The maximum IgM-antibody adsorption amount was 42.7 mg/g. IgM-antibody molecules could be repeatedly adsorbed and eluted without noticeable loss in the IgM-antibody adsorption amount. (C) 2010 Elsevier B.V. All rights reserved.

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