PubMed İndeksli Yayınlar Koleksiyonu / PubMed Indexed Publications Collection
Permanent URI for this collectionhttps://hdl.handle.net/11147/7645
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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: 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.