Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
Permanent URI for this collectionhttps://hdl.handle.net/11147/7148
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Article Citation - Scopus: 3Development of Chrono-Spectral Gold Nanoparticle Growth Based Plasmonic Biosensor Platform(Elsevier, 2024) Sözmen, Alper Baran; Elveren, Beste; Erdoğan, Duygu; Mezgil, Bahadır; Baştanlar, Yalın; Yıldız, Ümit Hakan; Arslan Yıldız, AhuPlasmonic sensor platforms are designed for rapid, label-free, and real-time detection and they excel as the next generation biosensors. However, current methods such as Surface Plasmon Resonance require expertise and well-equipped laboratory facilities. Simpler methods such as Localized Surface Plasmon Resonance (LSPR) overcome those limitations, though they lack sensitivity. Hence, sensitivity enhancement plays a crucial role in the future of plasmonic sensor platforms. Herein, a refractive index (RI) sensitivity enhancement methodology is reported utilizing growth of gold nanoparticles (GNPs) on solid support and it is backed up with artificial neural network (ANN) analysis. Sensor platform fabrication was initiated with GNP immobilization onto solid support; immobilized GNPs were then used as seeds for chrono-spectral growth, which was carried out using NH2OH at varied incubation times. The response to RI change of the platform was investigated with varied concentrations of sucrose and ethanol. The detection of bacteria E.coli BL21 was carried out for validation as a model microorganism and results showed that detection was possible at 102 CFU/ml. The data acquired by spectrophotometric measurements were analyzed by ANN and bacteria classification with percentage error rates near 0% was achieved. The proposed LSPR-based, label-free sensor application proved that the developed methodology promises utile sensitivity enhancement potential for similar sensor platforms. © 2024 The Author(s)Article Citation - WoS: 3Citation - Scopus: 4Gene Cloning, Heterologous Expression and Biochemical Characterization of a Novel Extracellular Lipase From Rhizopus Oryzae Ku45(National Institute of Genetic Engineering and Biotechnology, 2020) Arslanoğlu, Alper; Çil, ÇağlarBackground: Lipases secreted front various Rhizopus oryzae strains were previously expressed in Escherichia coli, Pichia pastoris, and Saccharomyces cerevisiae and was shown to have distinct activities in response to different temperatures, metal ions, organic solvents, and specific substrates. However, until now, no other research biochemically characterized the functions of extracellular pro-lipase in a novel Rhizopus oryzae KU45. Objectives: Characterization of a novel extracellular lipase front fungus R. orvzae KU45 after heterologous expression in E. coli BL21 (DE3) strain. Materials and Methods: An extracellular lipase producing fungus was isolated from a soil sample and identified as a strain of R. oryzae by partial 18S rRNA gene sequencing. It was named as R. oryzae KU45. The lipase gene of KU45 was cloned into pET-28a expression vector and expressed in E. coli as inclusion bodies. The recombinant lipase was purified, refolded and characterized. Results: The lipase exhibited maximum activity at 45 degrees C, at slightly alkaline pH. It showed a broad substrate specificity acting on p-nitrophenyl esters with C-8-C-16 acyl groups as substrates and, many of the organic solvents and metal ions tested did not have any adverse effects on the enzyme activity. Conclusions: High stability, broad substrate specificity and activity at mesophilic temperatures in the presence of organic solvents, and metal ions make the extracellular lipase of KU45 a candidate for various biotechnological applications.Article Citation - WoS: 16Citation - Scopus: 18Bacterial Detection Using Bacteriophages and Gold Nanorods by Following Time-Dependent Changes in Raman Spectral Signals(Informa Healthcare, 2018) Moghtader, Farzaneh; Tomak, Aysel; Zareie, Hadi M.; Pişkin, ErhanThis study attemps to develop bacterial detection strategies using bacteriophages and gold nanorods (GNRs) by Raman spectral analysis. Escherichia coli was selected as the target and its specific phage was used as the bioprobe. Target bacteria and phages were propagated/purified by traditional techniques. GNRs were synthesized by using hexadecyltrimethyl ammonium bromide (CTAB) as stabilizer. A two-step detection strategy was applied: Firstly, the target bacteria were interacted with GNRs in suspensions, and then they were dropped onto silica substrates for detection. It was possible to obtain clear surface-enchanced Raman spectroscopy (SERS) peaks of the target bacteria, even without using phages. In the second step, the phage nanoemulsions were droped onto the bacterial-GNRs complexes on those surfaces and time-dependent changes in the Raman spectra were monitored at different time intervals upto 40 min. These results demonstrated that how one can apply phages with plasmonic nanoparticles for detection of pathogenic bacteria very effectively in a quite simple test.Article Citation - WoS: 18Citation - Scopus: 23A Thermophilic ?-L From Geobacillus Vulcani Gs90: Heterologous Expression, Biochemical Characterization, and Its Synergistic Action in Fruit Juice Enrichment(Springer, 2018) İlgü, Hüseyin; Sürmeli, Yusuf; Şanlı Mohamed, Gülşahα-l-Arabinofuranosidases with an orchestral action of xylanolytic enzymes degrades the xylan in plant cell wall. In this study, heterologous expression, biochemical characterization, and synergistic action of α-l-Arabinofuranosidase from previously identified.Geobacillus vulcani GS90 (GvAbf) was investigated. The recombinant α-l-Arabinofuranosidase was overexpressed in Escherichia coli BL21 (λDE) and purified via His-tag Ni-affinity and size-exclusion chromatography. Optimum activity of the purified α-l-Arabinofuranosidase was obtained at pH 5 and at 70 °C. The GvAbf was active in a broad pH and temperature ranges; pH 4–9 and 30–90 °C, respectively. In addition, it retained most of its activity after an hour incubation at 70 °C and remained relatively stable at pH 3–6. GvAbf was quite stable against various metal ions. The kinetic parameters of GvAbf was obtained as Vmax and Km; 200 U/mg and 0.2 mM with p-nitrophenyl-α-l-arabinofuranoside and 526 U/mg and 0.1 mM with sugar beet arabinan, respectively. The synergistic action of GvAbf was studied with commercially available xylanase on juice enrichment of apples, grapes, oranges, and peaches. The best juice enrichment in terms of clarity, reducing sugar content, and yield, was achieved with GvAbf and xylanase together compared to treatment with xylanase and GvAbf alone in all fruits. The treatment with GvAbf and xylanase together lead to an increased juice yield by 26.56% (apple), 30.88% (grape), 40.00% (orange) and 32.20% (peach) as well as having a significant effect on juice clarity by an increase of % transmittance 47.26, 25.98, 41.77, and 44.97, respectively. The highest reducing sugar level of fruit juices also obtained with GvAbf and xylanase together compared to treatment with xylanase and GvAbf alone in all types of fruits. GvAbf and xylanase together as simultaneous synergistic manner may have an exciting potential for application in fruit juice processing.Article Citation - WoS: 81Citation - Scopus: 115Effects of Ultraviolet Light Emitting Diodes (leds) on Microbial and Enzyme Inactivation of Apple Juice(Elsevier Ltd., 2017) Pelvan Akgün, Merve; Ünlütürk, SevcanIn this study, the effects of Ultraviolet light-emitting diodes (UV-LEDs) on the inactivation of E. coli K12 (ATCC 25253), an indicator organism of E. coli O157:H7, and polyphneoloxidase (PPO) in cloudy apple juice (CAJ) were investigated. The clear (AJ) and cloudy apple juice were exposed to UV rays for 40 min by using a UV device composed of four UV-LEDs with peak emissions at 254 and 280 nm and coupled emissions as follows: 254/365, 254/405, 280/365, 280/405 and 254/280/365/405 nm. UV-LEDs at 254 nm achieved 1.6 ± 0.1 log10 CFU/mL inactivation of E. coli K12 at UV dose of 707.2 mJ/cm2. The highest inactivation of E. coli K12 (2.0 ± 0.1 log10 CFU/mL and 2.0 ± 0.4 log10 CFU/mL) was achieved when the cloudy apple juice was treated with both 280 nm and 280/365 nm UV-LEDs. For clear apple juice the highest inactivation 4.4 log10 CFU/mL obtained for E. coli K12 was achieved using 4 lamps emitting light at 280 nm for 40 min exposure time. For the same treatment time, the experiments using a combination of lamps emitting light at 280 and 365 nm (2lamp/2lamp) were resulted in 3.9 ± 0.2 log10 CFU/mL reductions. UV-A and UV-C rays in combination showed a better inactivation effect on PPO than UV-C rays used separately. Residual activity of PPO in CAJ was reduced to 32.58% when treated with UV-LED in combination of UV-C (280 nm) and UV-A (365 nm) rays. Additionally, the total color change (ΔE) of CAJ subjected to combined UV-LED irradiation at 280/365 nm was the lowest compared to other studied processing conditions. This study provides key implications for the future application of UV-LEDs to fruit juice pasteurization.Article Citation - WoS: 45Citation - Scopus: 47Impedimetric Detection of Pathogenic Bacteria With Bacteriophages Using Gold Nanorod Deposited Graphite Electrodes(Royal Society of Chemistry, 2016) Moghtader, Farzaneh; Çongur, Gülşah; Zareie, Hadi M.; Erdem, Arzum; Pişkin, ErhanElectrochemical impedance spectroscopy (EIS) is applied for the detection of bacteria using bacteriophages as a bioprobe together with gold nanorods (GNRs). Escherichia coli-E. coli K12 was used as a model target bacteria and also for the propagation of its specific T4-phages. Gold nanorods (GNRs) were synthesized via a two-step protocol and characterized using different techniques. EIS measurements were conducted in an electrochemical cell consisting of a three electrode system. Single-use pencil graphite electrodes (PGE) were modified by the physical adsorption of GNRs to increase their interfacial conductivity and therefore sensitivity for impedimetric measurements. Therefore, interfacial charge-transfer resistance values (Rct) sharply decreased after GNRs deposition. Phages were adsorbed on these electrodes via a simple incubation protocol at room temperature, which resulted in an increase in Rct values, which was concluded to be as a result of nonconductive phage layers. These phage-carrying GNRs-PGEs were used for impedimetric detection of the target bacteria, E. coli. Significant increases at the Rct values were observed which were attributed to the insulation effects of the adsorbed bacterial layers. This increase was even more when the bacterial concentrations were higher. In the case of the non-target bacteria Staphylococcus aureus (S. aureus), conductivity noticeable decreases (due to nonspecific adsorption). However, in the case of E. coli, the Rct value increase is time dependent and reaches maximum in about 25-30 min, then decreases gradually as a result of bacterial lysis due to phage invasion on the electrode surfaces. In contrast, there were no time dependent changes with the non-target bacteria S. aureus (no infection and no lytic activity). It is concluded that the target bacteria could be detected using this very simple and inexpensive detection protocol with a minimum detection limit of 103 CFU mL-1 in approximately 100 μL bacterial suspension.Article Citation - WoS: 22Citation - Scopus: 33Uv-C Irradiation of Freshly Squeezed Grape Juice and Modeling Inactivation Kinetics(John Wiley and Sons Inc., 2014) Ünlütürk, Sevcan; Atılgan, Mehmet ReşatUV inactivation kinetics of freshly squeezed turbid white grape juice (FSTGJ) treated with an annular flow UV reactor by applying UV dosages ranging from 0 to 116.7J/mL, at three different flow rates (0.90, 1.75 and 3.70mL/s), were modeled by using log-linear, Weibull, Hom and modified Chick-Watson models. FSTGJ was circulated five times in the UV system, i.e., UV exposure time was 20.33min during processing. The populations of Escherichia coli K-12, lactic acid bacteria (LAB) and foodborne yeasts were reduced by 3.759, 4.133 and 1.604log cfu/mL, respectively, after exposure to UV dosage of 116.7J/mL at the lowest flow rate. The inactivation kinetics of foodborne yeasts were best described by the modified Chick-Watson model, with the least root mean squared error (RMSE=0.001, R2=0.999). Besides, the inactivation kinetics of E.coli K-12 and LAB were best fitted by Weibull model (R2=0.999). Additionally, when the UV exposure time was increased up to 32.5min (i.e., eight cycles), UV-C treatment of FSTGJ resulted in 5.341log cfu/mL reduction in E.coli K-12, which meets the Food and Drug Administration requirement of a 5log reduction of microorganisms in fruit juices. Practical Applications Consumer demand for high-quality fruit juice with fresh-like characteristics has markedly expanded in recent years. UV-C irradiation is a nonthermal method and allows the processing of fruit juices with a minimal or no changes in flavor, essential nutrients and vitamins. Although thermal pasteurization is the most convenient way of increasing the shelf life of fruit juices, it causes a "cook taste" in grape juice. So, in this study, the application of UV-C irradiation to process grape juice was investigated. The shape of the microbial inactivation curve is sigmoidal in UV treatment. Therefore, different kinetic models (e.g., log-linear, Weibull, Hom and modified Chick-Watson) are applied to describe the inactivation kinetics of Escherichia coli K-12, lactic acid bacteria and foodborne yeasts. Kinetic parameters (e.g., k and D) and models can be used for the development of UV-C irradiation process to ensure microbial safety in juice products.Article Citation - WoS: 124Citation - Scopus: 144Use of Uv-C Radiation as a Non-Thermal Process for Liquid Egg Products (lep)(Elsevier Ltd., 2008) Ünlütürk, Sevcan; Atılgan, Mehmet Reşat; Baysal, Ayşe Handan; Tarı, CananThe efficacy of short wave ultraviolet light (UV-C) as a non-thermal process for liquid egg products (LEP) was investigated. Non-pathogenic Escherichia coli strain (ATCC 8739), which shows lower sensitivity to UV-C light than E. coli O157:H7 and Salmonella typhimurium, was chosen as a target microorganism. The inactivation of UV resistant strain of E. coli in LEP was examined by evaluating the effects of depth of liquid food medium (0.5, 0.3 and 0.153 cm), UV light intensity (1.314, 0.709 and 0.383 mW/cm2) and exposure time (0, 5, 10, and 20 min) by using a collimated beam apparatus. The best reduction (>2-log) was achieved in liquid egg white (LEW) when the fluid depth and UV intensity were 0.153 cm and 1.314 mW/cm2, respectively. Maximum inactivation was 0.675-log CFU/ml in liquid egg yolk (LEY) and 0.316-log CFU/ml in liquid whole egg (LWE) at the same conditions. The kinetics of UV inactivation of E. coli in LEP was nonlinear. Our results emphasize that UV-C radiation can be used as a pre-treatment process or combined with mild heat treatment to reduce the adverse effects of thermal pasteurization of LEP.