Browsing by Subject "Escherichia coli"

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Open Access
Antibacterial electrospun nanofibers from triclosan/cyclodextrin inclusion complexes
(Elsevier, 2014) Celebioglu A.; Umu, O. C. O.; Tekinay, T.; Uyar, Tamer
The electrospinning of nanofibers (NF) from cyclodextrin inclusion complexes (CD-IC) with an antibacterial agent (triclosan) was achieved without using any carrier polymeric matrix. Polymer-free triclosan/CD-IC NF were electrospun from highly concentrated (160% CD, w/w) aqueous triclosan/CD-IC suspension by using two types of chemically modified CD; hydroxypropyl-beta-cyclodextrin (HPβCD) and hydroxypropyl-gamma-cyclodextrin (HPγCD). The morphological characterization of the electrospun triclosan/CD-IC NF by SEM elucidated that the triclosan/HPβCD-IC NF and triclosan/HPγCD-IC NF were bead-free having average fiber diameter of 520±250nm and 1100±660nm, respectively. The presence of triclosan and the formation of triclosan/CD-IC within the fiber structure were confirmed by 1H-NMR, FTIR, XRD, DSC, and TGA studies. The initial 1:1molar ratio of the triclosan:CD was kept for triclosan/HPβCD-IC NF after the electrospinning and whereas 0.7:1molar ratio was observed for triclosan/HPγCD-IC NF and some uncomplexed triclosan was detected suggesting that the complexation efficiency of triclosan with HPγCD was lower than that of HPβCD. The antibacterial properties of triclosan/CD-IC NF were tested against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria. It was observed that triclosan/HPβCD-IC NF and triclosan/HPγCD-IC NF showed better antibacterial activity against both bacteria compared to uncomplexed pure triclosan.
Open Access
Antibacterial electrospun poly(lactic acid) (PLA) nanofibrous webs incorporating triclosan/cyclodextrin inclusion complexes
(2013) Kayaci F.; Umu O.C.O.; Tekinay, T.; Uyar, T.
Solid triclosan/cyclodextrin inclusion complexes (TR/CD-IC) were obtained and then incorporated in poly(lactic acid) (PLA) nanofibers via electrospinning. α-CD, β-CD, and γ-CD were tested for the formation of TR/CD-IC by a coprecipitation method; however, the findings indicated that α-CD could not form an inclusion complex with TR, whereas β-CD and γ-CD successfully formed TR/CD-IC crystals, and the molar ratio of TR to CD was found to be 1:1. The structural and thermal characteristics of TR/CD-IC were investigated by 1H NMR, FTIR, XRD, DSC, and TGA studies. Then, the encapsulation of TR/β-CD-IC and TR/γ-CD-IC in PLA nanofibers was achieved. Electrospun PLA and PLA/TR nanofibers obtained for comparison were uniform, whereas the aggregates of TR/CD-IC crystals were present and distributed within the PLA fiber matrix as confirmed by SEM and XRD analyses. The antibacterial activity of these nanofibrous webs was investigated. The results indicated that PLA nanofibers incorporating TR/CD-IC showed better antibacterial activity against Staphylococcus aureus and Escherichia coli bacteria compared to PLA nanofibers containing only TR without CD-IC. Electrospun nanofibrous webs incorporating TR/CD-IC may be applicable in active food packaging due to their very high surface area and nanoporous structure as well as efficient antibacterial property. © 2013 American Chemical Society.
Open Access
Antibacterial electrospun zein nanofibrous web encapsulating thymol/cyclodextrin-inclusion complex for food packaging
(Elsevier, 2017-10) Aytac Z.; Ipek, S.; Durgun, Engin; Tekinay, T.; Uyar, Tamer
Thymol (THY)/γ-Cyclodextrin(γ-CD) inclusion complex (IC) encapsulated electrospun zein nanofibrous webs (zein-THY/γ-CD-IC-NF) were fabricated as a food packaging material. The formation of THY/γ-CD-IC (1:1 and 2:1) was proved by experimental (X-ray diffraction (XRD), thermal gravimetric analysis (TGA), 1H NMR) and computational techniques. THY/γ-CD-IC (2:1) exhibited higher preservation rate and stability than THY/γ-CD-IC (1:1). It is worth mentioning that zein-THY/γ-CD-IC-NF (2:1) preserved much more THY as observed in TGA and stability of THY/γ-CD-IC (2:1) was higher, as shown by a modelling study. Therefore, much more THY was released from zein-THY/γ-CD-IC-NF (2:1) than zein-THY-NF and zein-THY/γ-CD-IC-NF (1:1). Similarly, antibacterial activity of zein-THY/γ-CD-IC-NF (2:1) was higher than zein-THY-NF and zein-THY/γ-CD-IC-NF (1:1). It was demonstrated that zein-THY/γ-CD-IC-NF (2:1) was most effective in inhibiting the growth of bacteria on meat samples. These webs show potential application as an antibacterial food packaging material.
Open Access
Electrospinning of polymer-free cyclodextrin/geraniol-inclusion complex nanofibers: enhanced shelf-life of geraniol with antibacterial and antioxidant properties
(Royal Society of Chemistry, 2016) Aytac Z.; Yildiz, Z. I.; Kayaci-Senirmak, F.; Keskin, S. N. O.; Tekinay, T.; Uyar, Tamer
Free-standing nanofibrous webs of cyclodextrin/geraniol-inclusion complex (CD/geraniol-IC-NF) showing antibacterial, antioxidant activity and slow release of geraniol were developed as flavour/fragrance releasing materials via electrospinning. The electrospinning of CD/geraniol-IC-NFs with uniform and bead-free morphology was achieved without using a polymer matrix. Three types of CDs modified with hydroxypropyl and methyl groups (HPβCD, MβCD, and HPγCD) were used to obtain CD/geraniol-IC-NFs. The polymer-free CD/geraniol-IC-NFs allow us to attain much higher geraniol loading (∼11%, w/w) when compared to electrospun polymeric nanofibers containing CD/geraniol-IC (∼5%, w/w). Geraniol has a volatile nature, yet, a significant amount of geraniol (∼60-90%) was preserved in CD/geraniol-IC-NFs due to the complexation, whereas evaporation of geraniol was unavoidable for polymeric nanofibers incorporating geraniol without cyclodextrin. Short-term (3 h) temperature dependent release (37 °C, 50 °C, and 75 °C) and long-term open air (50 days, at RT) release tests revealed that MβCD/geraniol-IC-NF released less geraniol compared to HPβCD/geraniol-IC-NF and HPγCD/geraniol-IC-NF, indicating that much stronger inclusion complexation was formed between MβCD and geraniol. The release of geraniol from CD/geraniol-IC-NFs prevented the colonization of Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria to a great extent, as observed in the antibacterial activity results. It was observed that CD/geraniol-IC-NFs had higher antioxidant activity compared to pure geraniol due to the solubility increase. In brief, the results reported here may open a new door to enhance the performance of essential oils and flavour/fragrances, to preserve volatile compounds from evaporation and to better understand the potential of CD/IC-NFs as carrier systems for guest compounds in the food, cosmetic and household cleaning industries.
Open Access
Fast-dissolving, prolonged release, and antibacterial cyclodextrin/limonene-inclusion complex nanofibrous webs via polymer-free electrospinning
(American Chemical Society, 2016) Aytac Z.; Yildiz, Z. I.; Kayaci-Senirmak, F.; S. Keskin, N. O.; Kusku, S. I.; Durgun, Engin; Tekinay, T.; Uyar, Tamer
We have proposed a new strategy for preparing free-standing nanofibrous webs from an inclusion complex (IC) of a well-known flavor/fragrance compound (limonene) with three modified cyclodextrins (HPβCD, MβCD, and HPγCD) via electrospinning (CD/limonene-IC-NFs) without using a polymeric matrix. The experimental and computational modeling studies proved that the stoichiometry of the complexes was 1:1 for CD/limonene systems. MβCD/limonene-IC-NF released much more limonene at 37, 50, and 75 °C than HPβCD/limonene-IC-NF and HPγCD/limonene-IC-NF because of the greater amount of preserved limonene. Moreover, MβCD/limonene-IC-NF has released only 25% (w/w) of its limonene, whereas HPβCD/limonene-IC-NF and HPγCD/limonene-IC-NF released 51 and 88% (w/w) of their limonene in 100 days, respectively. CD/limonene-IC-NFs exhibited high antibacterial activity against E. coli and S. aureus. The water solubility of limonene increased significantly and CD/limonene-IC-NFs were dissolved in water in a few seconds. In brief, CD/limonene-IC-NFs with fast-dissolving character enhanced the thermal stability and prolonged the shelf life along with antibacterial properties could be quite applicable in food and oral care applications.
Open Access
Functional role of the noncatalytic domains of elongation factor tu in the interactions with ligands
(1998) Cetin, R.; Anborgh, P. H.; Cool, R. H.; Parmeggiani, A.
Elongation factor (EF) Tu from Escherichia coli contains three domains, of which domain 1 (N-terminal domain) harbors the site for nucleotide binding and GTP hydrolysis. To analyze the function of domains 2 [middle (M) domain] and 3 [C-terminal (C) domain], EF-Tu(AM) and EF-Tu(δC) were engineered as GST-fused products and purified. Circular dichroism and thermostability showed that both constructs have conserved organized structures. Though inactive in poly(Phe) synthesis the two constructs could bind GDP and GTP with comparable micromolar affinities. Therefore, like the isolated N- terminal domain, they had lost a typical feature of EF-Tu, the ≤ 100 times stronger affinity for GDP than for GTP. EF-Tu(ΔM) and EF-Tu(AC) had an intrinsic GTPase activity comparable to that of wild-type EF-Tu. Ribosomes did not stimulate the GTPase activity of either factor, while kirromycin increased the GTPase activity of both constructs, particularly of EF-Tu(ΔC), to a level, however, much lower than that of the intact molecule. The interaction with aa-tRNA of both mutants was ≤90% reduced. As a major result, their GDP-bound form could efficiently respond to EF-Ts. All four EF- Tu-specific antibiotics [kirromycin, pulvomycin, GE2270 A (=MDL 62 879), and enacyloxin IIa] retarded significantly the dissociation of EF-Tu(ΔC)·GTP, showing the same kind of effect as on EF-Tu·GTP, but they were little active on EF-Tu(ΔM)·GTP. Like EF-Tu(ΔC)·GTP, EF-Tu(ΔM)·GTP was, however, able to bind efficiently kirromycin and enacyloxin IIa, as determined via competition with EF-Ts. Together, these results enlight selective functions of domains 2 and 3, particularly toward the interaction with EF-Ts and antibiotics, and emphasize their functional cooperativity for an efficient interaction of EF-Tu with ribosomes and aa-tRNA and for maintaining the differential affinity for GTP and GDP.
Open Access
Genetically encoded conductive protein nanofibers secreted by engineered cells
(Royal Society of Chemistry, 2017-06) Kalyoncu, E.; Ahan, R. E.; Olmez, T. T.; Safak Seker, U. O.
Bacterial biofilms are promising tools for functional applications as bionanomaterials. They are synthesized by well-defined machinery, readily form fiber networks covering large areas, and can be engineered for different functionalities. In this work, bacterial biofilms have been engineered for use as conductive biopolymers to interface with electrodes and connect bacterial populations to electronic gadgets. Bacterial biofilms are designed with different conductive peptide motifs, as the aromatic amino acid content of fused peptide motifs has been suggested to contribute to electronic conductivity by influencing monomer stacking behavior. To select the best candidates for constructing conductive peptide motifs, conductivity properties of aromatic amino acids are measured using two different fiber scaffolds, an amyloid-like fiber (ALF) forming peptide, and the amyloidogenic R5T peptide of CsgA protein. Three repeats of aromatic amino acids are added to fiber-forming peptide sequences to produce delocalized π clouds similar to those observed in conductive polymers. Based on the measurements, tyrosine and tryptophan residues provide the highest conductivity. Therefore, the non-conductive E. coli biofilm is switched into a conductive form by genetically inserted conductive peptide motifs containing different combinations of tyrosine and tryptophan. Finally, synthetic biofilm biogenesis is achieved with conductive peptide motifs using controlled biofilm production. Conductive biofilms on living cells are formed for bioelectronics and biosensing applications.
Open Access
Genetically-tunable mechanical properties of bacterial functional amyloid nanofibers
(American Chemical Society, 2017) Abdelwahab, M. T.; Kalyoncu, E.; Onur, T.; Baykara, M. Z.; Seker U.O.S.
Bacterial biofilms are highly ordered, complex, dynamic material systems including cells, carbohydrates, and proteins. They are known to be resistant against chemical, physical, and biological disturbances. These superior properties make them promising candidates for next generation biomaterials. Here we investigated the morphological and mechanical properties (in terms of Young’s modulus) of genetically-engineered bacterial amyloid nanofibers of Escherichia coli (E. coli) by imaging and force spectroscopy conducted via atomic force microscopy (AFM). In particular, we tuned the expression and biochemical properties of the major and minor biofilm proteins of E. coli (CsgA and CsgB, respectively). Using appropriate mutants, amyloid nanofibers constituting biofilm backbones are formed with different combinations of CsgA and CsgB, as well as the optional addition of tagging sequences. AFM imaging and force spectroscopy are used to probe the morphology and measure the Young’s moduli of biofilm protein nanofibers as a function of protein composition. The obtained results reveal that genetically-controlled secretion of biofilm protein components may lead to the rational tuning of Young’s moduli of biofilms as promising candidates at the bionano interface.
Open Access
A highly sensitive and specific enzyme-linked immunosorbent assay of antibodies to hepatitis C virus
(2000) Eroğlu, C.; Yıldız, E.; Öztürk, M.; Pınarbaşı, E.
In this study, a 178 amino acids long portion of the hepatitis C virus (HCV) core gene was cloned, sequenced, expressed in Escherichia coli, and purified. The resulting antigen (C178) was tested with human sera enzyme-linked immunosorbent assay (ELISA) in order to assess its ability to diagnose HCV. It was shown by ELISA that 92% of the patients sera, diagnosed previously by a 3(rd) generation enzyme immunoassay (EIA) as HCV-positive, had antibodies against the C178 antigen. This antigen gave no false positive results when tested with anti-HCV-negative sera.
Open Access
Manipulation and control of collective behavior in active matter systems
(2016-10) Pinçe, Erçağ
Active matter systems consist of active constituents that transform energy into directed motion in a non-equilibrium setting. The interaction of active agents with each other and with their environment results in collective motion and emergence of long-range ordering. Examples to such dynamic behaviors in living active matter systems are pattern formation in bacterial colonies, ocking of birds and clustering of pedestrian crowds. All these phenomena stem from far-from-equilibrium interactions. The governing dynamics of these phenomena are not yet fully understood and extensively studied. In this thesis, we studied the role that spatial disorder can play to alter collective dynamics in a colloidal living active matter system. We showed that the level of heterogeneity in the environment in uences the long-range order in a colloidal ensemble coupled to a bacterial bath where the non-equilibrium forces imposed by the bacteria become pivotal to control switching between gathering and dispersal of colloids. Apart from studying environmental factors in a complex active matter system, we also focused on a new class of active particles, \bionic microswimmers", and their clustering behavior. We demonstrated that spherical bionic microswimmers which are fabricated by attaching motile E. coli bacteria on melamine particles can agglomerate in large colloidal structures. Finally, we observed the emergence of swimming clusters as a result of the collective motion of bionic microswimmers. Our results provide insights about statistical behavior and far-from-equilibrium interactions in an active matter system.
Open Access
Multivalent presentation of cationic peptides on supramolecular nanofibers for antimicrobial activity
(American Chemical Society, 2017) Beter, M.; Kara, H. K.; Topal, A. E.; Dana, A.; Tekinay, A. B.; Güler, Mustafa O.
Noncovalent and electrostatic interactions facilitate the formation of complex networks through molecular self-assembly in biomolecules such as proteins and glycosaminoglycans. Self-assembling peptide amphiphiles (PA) are a group of molecules that can form nanofibrous structures and may contain bioactive epitopes to interact specifically with target molecules. Here, we report the presentation of cationic peptide sequences on supramolecular nanofibers formed by self-assembling peptide amphiphiles for cooperative enhanced antibacterial activity. Antibacterial properties of self-assembled peptide nanofibers were significantly higher than soluble peptide molecules with identical amino acid sequences, suggesting that the tandem presentation of bioactive epitopes is important for designing new materials for bactericidal activity. In addition, bacteria were observed to accumulate more rapidly on peptide nanofibers compared to soluble peptides, which may further enhance antibacterial activity by increasing the number of peptide molecules interacting with the bacterial membrane. The cationic peptide amphiphile nanofibers were observed to attach to bacterial membranes and disrupt their integrity. These results demonstrate that short cationic peptides show a significant improvement in antibacterial activity when presented in the nanofiber form.
Open Access
One-step synthesis of size-tunable Ag nanoparticles incorporated in electrospun PVA/cyclodextrin nanofibers
(Pergamon Press, 2014) Celebioglu A.; Aytac Z.; Umu, O. C. O.; Dana, A.; Tekinay, T.; Uyar, Tamer
One-step synthesis of size-tunable silver nanoparticles (Ag-NP) incorporated into electrospun nanofibers was achieved. Initially, in situ reduction of silver salt (AgNO3) to Ag-NP was carried out in aqueous solution of polyvinyl alcohol (PVA). Here, PVA was used as reducing agent and stabilizing polymer as well as electrospinning polymeric matrix for the fabrication of PVA/Ag-NP nanofibers. Afterwards, hydroxypropyl-beta-cyclodextrin (HPβCD) was used as an additional reducing and stabilizing agent in order to control size and uniform dispersion of Ag-NP. The size of Ag-NP was ∼8 nm and some Ag-NP aggregates were observed for PVA/Ag-NP nanofibers, conversely, the size of Ag-NP decreased from ∼8 nm down to ∼2 nm within the fiber matrix without aggregation were attained for PVA/HPβCD nanofibers. The PVA/Ag-NP and PVA/HPβCD/Ag-NP nanofibers exhibited surface enhanced Raman scattering (SERS) effect. Moreover, antibacterial properties of PVA/Ag-NP and PVA/HPβCD/Ag-NP nanofibrous mats were tested against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria.
Open Access
Release and antibacterial activity of allyl isothiocyanate/β-cyclodextrin complex encapsulated in electrospun nanofibers
(Elsevier, 2014) Aytac Z.; Dogan, S.Y.; Tekinay, T.; Uyar, Tamer
Allyl isothiocyanate (AITC) is known as an efficient antibacterial agent but it has a very high volatility. Herein, AITC and AITC/β-cyclodextrin (CD)-inclusion complex (IC) incorporated in polyvinyl alcohol (PVA) nanofibers were produced via electrospinning. SEM images elucidated that incorporation of AITC and AITC/β-CD-IC into polymer matrix did not affect the bead-free fiber morphology of PVA nanofibers. 1H-NMR and headspace GC-MS analyses revealed that very low amount of AITC was remained in PVA/AITC-NF because of the rapid evaporation of AITC during the electrospinning process. Nevertheless, much higher amount of AITC was preserved in the PVA/AITC/β-CD-IC-NF due to the CD inclusion complexation. The sustained release of AITC from nanofibers was evaluated at 30°C, 50°C and 75°C via headspace GC-MS. When compared to PVA/AITC-NF, PVA/AITC/β-CD-IC-NF has shown higher antibacterial activity against Escherichia coli and Staphylococcus aureus due to the presence of higher amount of AITC in this sample which was preserved by CD-IC. © 2014 Elsevier B.V.
Open Access
Simultaneous photoinduced electron transfer and photoinduced CuAAC processes for antibacterial thermosets
(Elsevier, 2017) Oz, E.; Uyar, T.; Esen, H.; Tasdelen, M. A.
A combination of simultaneous photoinduced electron transfer and photoinduced CuAAC processes enables the in-situ preparation of antibacterial thermosets containing silver nanoparticles (AgNPs) in one-pot. Upon photolysis of photoinitator, the generated radicals not only reduce Cu(II) into Cu(I) activator to catalyst the CuAAC click reaction, but also simultaneously generate AgNPs from AgNO3 through electron transfer reaction. Due to their reduction potentials difference, the polymer matrix is formed before the formation of AgNPs, assisting to eliminate the agglomeration of them. The thermoset structures are confirmed by FT-IR and solubility tests, whereas the presence of AgNPs is proven by transmission electron microscopy with energy dispersive X-ray system analyzer. The samples containing 5 and 10% AgNPs exhibited strong inhibition zones, where all kinds of bacteria (gram-positive (Staphylococcus Aureus) and gram-negative (Escherichia Coli)) were killed in the surrounding of the film samples.
Open Access
Synthesis, characterization and antibacterial investigation of silver-copper nanoalloys
(2011) Taner, M.; Sayar, N.; Yulug I. G.; Süzer, Şefik
Ag-Cu nanoalloys were synthesized by chemical co-reduction of their metal salts in aqueous solution with hydrazine hydrate, in the presence of complexing agent and stabilizer, preventing the oxidation of copper, as revealed by XPS. Their antibacterial behavior was tested against Escherichia coli strains, attesting far better ability of the Ag-Cu compared to Ag-only nanoparticles.
Open Access
Synthetic biogenesis of bacterial amyloid nanomaterials with tunable inorganic-organic interfaces and electrical conductivity
(American Chemical Society, 2017) Seker U.O.S.; Chen, A. Y.; Citorik, R. J.; Lu, T. K.
Amyloids are highly ordered, hierarchal protein nanoassemblies. Functional amyloids in bacterial biofilms, such as Escherichia coli curli fibers, are formed by the polymerization of monomeric proteins secreted into the extracellular space. Curli is synthesized by living cells, is primarily composed of the major curlin subunit CsgA, and forms biological nanofibers with high aspect ratios. Here, we explore the application of curli fibers for nanotechnology by engineering curli to mediate tunable biological interfaces with inorganic materials and to controllably form gold nanoparticles and gold nanowires. Specifically, we used cell-synthesized curli fibers as templates for nucleating and growing gold nanoparticles and showed that nanoparticle size could be modulated as a function of curli fiber gold-binding affinity. Furthermore, we demonstrated that gold nanoparticles can be preseeded onto curli fibers and followed by gold enhancement to form nanowires. Using these two approaches, we created artificial cellular systems that integrate inorganic-organic materials to achieve tunable electrical conductivity. We envision that cell-synthesized amyloid nanofibers will be useful for interfacing abiotic and biotic systems to create living functional materials.