Bioengineering / Biyomühendislik
Permanent URI for this collectionhttps://hdl.handle.net/11147/4529
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Conference Object A Glucuronoxylan-Based Bio-Ink Development: Characterization and Application(Wiley, 2023) Yıldırım, Ömer; Arslan Yıldız, AhuBioprinting is a trending technique that enables the fabrication of threedimensional (3D) constructs in designed shapes and with desired properties. Bioinks are one of the most significant components of bioprinting and the successful fabrication of 3D bioprinted constructs mostly depends on the features of bioinks that would be used. New generation bioinks that are soft and viscous enough, printable under low pressure, stable in cell culture, and have fast gelation mechanisms are ideal to be used in current bioprinting techniques. Hydrocolloids have said features and have similar properties to native ECM structures. Hence bioinks that are developed from hydrocolloids can be utilized for mimicking of ECM structure of soft tissues. Polysaccharidebased hydrocolloids are ideal bioink candidates with their high waterholding capacity and biocompatibility. Here, a glucuronoxylanbased newgeneration bioink was developed, and its printability was evaluated for 3D bioprinting applications. The glucuronoxylanbased hydrocolloid was obtained by water extraction of quince seeds and its utilization in bioprinting was investigated. Bioink characterization was done by FTIR and mechanical analysis. Bioprinting parameters were optimized assessing uniformity, pore factor, and shape fidelity. Then, the characterization of bioprinted constructs was performed by pore angle measurement, waterholding capacity analysis, protein adsorption, and cell viability assays. Bioprinted structures have high mechanical strength, suitable protein adsorption behavior, and waterholding capacity as high as 20fold of its own weight, which is higher than other hydrogels that were used in soft tissue engineering. Moreover, the cell viability results of fibroblast cells in the bioink were high for longterm culture. In conclusion, findings show that the developed glucuronoxylanbased bioink is a biocompatible and promising bioink material for further tissue engineering applications.Article Citation - WoS: 3Citation - Scopus: 3Structural and Functional Analyses of Gh51 Alpha-L of Geobacillus Vulcani Gs90 Reveal Crucial Residues for Catalytic Activity and Thermostability(Wiley, 2022) Sürmeli, Yusuf; Şanlı Mohamed, GülşahAlpha-L-arabinofuranosidase (Abf) is of big interest in various industrial areas. Directed evolution is a powerful strategy to identify significant residues underlying Abf properties. Here, six active variants from GH51 Abf of Geobacillus vulcani GS90 (GvAbf) by directed evolution were overproduced, extracted, and analyzed at biochemical and structural levels. According to the activity and thermostability results, the most-active and the least-active variants were found as GvAbf51 and GvAbf52, respectively. GvAbf63 variant was more active than parent GvAbf by 20% and less active than GvAbf51. Also, the highest thermostability belonged to GvAbf52 with 80% residual activity after 1 h. Comparative sequence and structure analyses revealed that GvAbf51 possessed L307S displacement. Thus, this study suggested that L307 residue may be critical for GvAbf activity. GvAbf63 had H30D, Q90H, and L307S displacements, and H30 was covalently bound to E29 catalytic residue. Thus, H30D may decrease the positive effect of L307S on GvAbf63 activity, preventing E29 action. Besides, GvAbf52 possessed S215N, L307S, H473P, and G476C substitutions and S215 was close to E175 (acid–base residue). S215N may partially disrupt E175 action. Overall effect of all substitutions in GvAbf52 may result in the formation of the C–C bond between C171 and C213 by becoming closer to each other.Conference Object Immobilized Gold Nanoparticle Based Plasmonic Assay Platform for Biomolecule and Microorganism Detection(Wiley, 2021) Sözmen, Alper Baran; Arslan Yıldız, AhuPlasmonic sensors are suitable tools for study of molecular interactions. Localized Surface Plasmon Resonance (LSPR) based sensors detect spectral changes associated with intramolecular interactions between analyte molecules and recognition elements. Due to its labelfree and highly sensitive features, LSPR based methods have high potential for biosensing applications. In this study, we aim to develop a sensitive, labelfree, rapid and simple biosensing platform. For this purpose, a novel refractive index (RI) sensitivity enhancement methodology is proposed by immobilizing gold nanoparticles (GNPs) for platformbased LSPR. Fabrication of platform was carried out by GNP synthesis, immobilization of GNPs on polystyrene solid support, and growth of GNPs. Validation of response to RI changes of developed sensor platform was carried out by tests with varying concentrations of sucrose and ethanol. Then as a proofofconcept, detection ability and detection limit determination of E.coli BL21 (DE3) and protein Bovine Serum Albumin (BSA) was carried out. Adsorption of E.coli BL21 (DE3) via bulk interactions showed that the developed LSPR platform exhibit high enough binding affinity for bacteria detection, and was able to detect down to concentrations as low as 102 CFU/ml. Immune capturing of BSA via antiBSA antibody showed that the developed LSPR platform was able to detect BSA protein–antibody interaction down to 10 µM concentration range.Article Citation - WoS: 6Citation - Scopus: 6Functional Characterization of a Novel Cyp119 Variant To Explore Its Biocatalytic Potential(Wiley, 2021) Sakallı, Tuğçe; Sürmeli, Nur BaşakBiocatalysts are increasingly applied in the pharmaceutical and chemical industry. Cytochrome P450 enzymes (P450s) are valuable biocatalysts due to their ability to hydroxylate unactivated carbon atoms using molecular oxygen. P450s catalyze reactions using nicotinamide adenine dinucleotide phosphate (NAD(P)H) cofactor and electron transfer proteins. Alternatively, P450s can utilize hydrogen peroxide (H2O2) as an oxidant, but this pathway is inefficient. P450s that show higher efficiency with peroxides are sought after in industrial applications. P450s from thermophilic organisms have more potential applications as they are stable toward high temperature, high and low pH, and organic solvents. CYP119 is an acidothermophilic P450 from Sulfolobus acidocaldarius. In our previous study, a novel T213R/T214I (double mutant [DM]) variant of CYP119 was obtained by screening a mutant library for higher peroxidation activity utilizing H2O2. Here, we characterized the substrate scope; stability toward peroxides; and temperature and organic solvent tolerance of DM CYP119 to identify its potential as an industrial biocatalyst. DM CYP119 displayed higher stability than wild-type (WT) CYP119 toward organic peroxides. It shows higher peroxidation activity for non-natural substrates and higher affinity for progesterone and other bioactive potential substrates compared to WT CYP119. DM CYP119 emerges as a new biocatalyst with a wide range of potential applications in the pharmaceutical and chemical industry.Article Citation - WoS: 2Citation - Scopus: 2Cloning, Expression, and Characterization of a Novel Sericin-Like Protein(Wiley, 2022) Bostan, Fatmanur; Sürmeli, Nur BaşakSilk consists of two proteins called fibroin and sericin. While fibroin is used in the textile industry and has various biomaterial applications, sericin has been considered as waste material until recently. Sericin is a multicomponent protein and it has important properties such as biocompatibility, biodegradability, cryoprotectivity, and antioxidant. Sericin from silkworm cocoons can be obtained by chemical, enzymatic, and heat treatment methods. However, sericin obtained with these treatment methods is not of consistent and high quality. Moreover, the exposure of sericin to harsh conditions during extraction leads to inconsistencies in the composition and structure of the sericin obtained. The inconsistencies in sericin structure and composition decrease application of sericin as a biomaterial. Here, we produce a sericin-like protein (Ser4mer) with native sequence of sericin encoding four repeats of the conserved 38 amino acid motif recombinantly in Escherichia coli and characterize its structural properties. Ser4mer protein shows similar structure to native sericin and higher solubility than previously obtained recombinant sericin-like proteins. Recombinant production of a soluble sericin-like protein will significantly expand its applications as a biomaterial. In addition, recombinant production of silk proteins will allow us to understand sequence-structure relationships in these proteins.Conference Object Evaluation of Anti-Inflammatory Potential and Lc-ms/Ms Analysis of Different Standards(Wiley, 2018) Sözmen, Alper Baran; Canbay, E.; Yıldırım Sözmen, Eser; Övez, BikemStandardization of a LC-MS/MS method for phenolic compound content of biomass originated extracts was aimed in this study, Chlorella miniata extracts, which were cultivated solely for this study, were used for this purpose. The first step of the study was to compare external standard method, added standard method, and conventional standard method the qualitative identification of phenolic compounds of the extracts were performed by using LC-MS/MS system.