Bilgilendirme: Kurulum ve veri kapsamındaki çalışmalar devam etmektedir. Göstereceğiniz anlayış için teşekkür ederiz.

Scopus İndeksli Yayınlar Koleksiyonu

Permanent URI for this collectionhttps://hdl.handle.net/20.500.14901/696

Browse

Browsing Scopus İndeksli Yayınlar Koleksiyonu by Author "Abul, Nurgul"

Filter results by typing the first few letters
Now showing 1 - 2 of 2
  • Thumbnail Image
    Article
    Citation - WoS: 11
    Citation - Scopus: 13
    Affinity-Based and In a Single Step Purification of Recombinant Horseradish Peroxidase A2a Isoenzyme Produced by Pichia Pastoris
    (Springer, 2023) Acar, Melek; Abul, Nurgul; Yildiz, Seyda; Taskesenligil, Ezgi Dag; Gerni, Serpil; Unver, Yagmur; Ozdemir, Hasan
    Horseradish peroxidase (HRP) is an oxidoreductase enzyme and oxidizes various inorganic and organic compounds. It has wide application areas such as immunological tests, probe-based test techniques, removal of phenolic pollutants from wastewater and organic synthesis. HRP is found in the root of the horseradish plant as a mixture of different isoenzymes, and it is very difficult to separate these enzymes from each other. In this regard, recombinant production is a very advantageous method in terms of producing the desired isoenzyme. This study was performed to produce HRP A2A isoenzyme extracellularly in Pichia pastoris and to purify this enzyme in a single step using a 3-amino-4-chloro benzohydrazide affinity column. First, codon-optimized HRP A2A gene was amplified and inserted into pPICZ alpha C. So, obtained pPICZ alpha C-HRPA2A was cloned in E. coli cells. Then, P. pastoris X-33 cells were transformed with linearized recombinant DNA and a yeast clone was cultivated for extracellular recombinant HRP A2A (rHRP A2A) enzyme production. Then, the purification of this enzyme was performed in a single step by affinity chromatography. The molecular mass of purified rHRP A2A enzyme was found to be about 40 kDa. According to characterization studies of the purified enzyme, the optimum pH and ionic strength for the rHRP A2A isoenzyme were determined to be 6.0 and 0.04 M, respectively, and o-dianisidine had the highest specificity with the lowest Km and Vmax values. Thus, this is an economical procedure to purify HRP A2A isoenzyme without time-consuming and laborious isolation from an isoenzyme mixture.
  • Thumbnail Image
    Article
    Citation - WoS: 7
    Citation - Scopus: 5
    A New Affinity Matrix Synthesized from Aminobenzohydrazide Derivatives for Purification of Lactoperoxidase Enzyme
    (Wiley-V C H Verlag GmbH, 2022) Kavaz, Neslihan Macit; Erel, Deniz; Korkmaz, Isil Nihan; Gerni, Serpil; Abul, Nurgul; Bayrak, Songul; Ozdemir, Hasan
    In this study, a new affinity process was developed for the purification of Lactoperoxidase with synthesized sixteen aminobenzohydrazide derivatives. For this purpose, ligands were covalently bound to CNBr-activated Sepharose-4B-L-tyrosine matrix and affinity columns were prepared, and LPO was purified in one step with high yield and purity. Among all synthesized molecules, the 4-amino-3-bromo-2-methylbenzohydrazide molecule had a high usable potential in the purification of Lactoperoxidase from mammalian milk. Lactoperoxidase was purified 411.8 times with a yield of 17.38 % from goat milk, 187.25 times with a yield of 9.72 % buffalo milk, 2772.4 times with a yield of 18.98 % from bovine milk, and 1246.65 times with a yield of 4.43 % from sheep milk. It was demonstrated for the first time that aminobenzohydrazide molecules could be used as ligands in the purification of Lactoperoxidase enzyme.