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Enzyme Immobilization and Optimization on Polymeric Nanospheres

Yıl 2019, Cilt: 14 Sayı: 1, 97 - 104, 31.05.2019
https://doi.org/10.29233/sdufeffd.479246

Öz

Enzymes are important materials
that are used in many industries and they have a very high cost. Although the
conditions for the enzymes in catalytic reactions are appropriate, enzymes can
easily lose their activity in the presence of high temperature, high or low pH
and organic / inorganic solvents. With the immobilization process, it is
possible to ensure that the enzymes are economically usable. For this purpose, the
glucose oxidase (GOx) enzyme was immobilized to aminomethyl polystyrene (APS)
nanospheres and the optimization parameters (pH, temperature, reusability
number, storage stability, substrate concentration) were determined. Michaelis-Menten equation for free and
immobilized enzyme and Km
and Vmax values were
calculated. In particular, it has been found that the substrate of the immobilized
enzyme containing Pt(IV) ion shows a high affinity for the substrate. After 12
months, immobilized enzymes were found to maintain about 85% of their activity.

Kaynakça

  • N. Kurnaz Yetim, “Ferrosen içeren yeni Pt(II), Pt(IV) dendrimerlerinin sentezi ve immobilizasyonda kullanımı,” Doktora Tezi, Fen Bil. Enst., Gazi Üni., Ankara, Türkiye, 2016.
  • A. Wiseman, “Handbook of Enzymes Biotechnology (Book style with paper title and editor),” in The application of enzymes in industry, 2nd ed., E. Horwood, Ed. Chichester, UK, 1987, pp. 274-373.
  • E. Hasanoğlu Özkan, N. Kurnaz Yetim, D. Nartop, N. Sarı, “Influence of load on the recycling stability of nanospheres attached platinium ion for determination of glucose,” J. Ind. Eng. Chem., 25, 180-185, 2015.
  • S. Dönmez, F. Arslan, N. Sarı, E. Hasanoğlu Özkan, H. Arslan, “Glucose biosensor based on immobilization of glucose oxidase on a carbon paste electrode modified with microspheres attached L-glycine,” Biotechnol. Appl. Biochem., 64 (5), 745-753, 2017.
  • X. Wang, Y. Zhang, C. E. Banks, Q. Chen and X. Ji, “Non-enzymatic amperometric glucose biosensor based on nickel hexacyanoferrate nanoparticle film modified electrodes,” Colloids Surf. B, 78, 363-366, 2010.
  • H. Öğütcü, N. Kurnaz Yetim, E. Hasanoğlu Özkan, O. Eren, G. Kaya, N. Sarı, and A. Dişli, “Nanospheres Caped Pt(II) and Pt (IV): Synthesis and Evaluation as Antimicrobial and Antifungal Agent,” Pol. J. Chem., 19, 74-80, 2017.
  • P. Trinder, “Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor,” Ann. Clin. Biochem., 6, 24-27, 1969.
  • V. Leskovac, S. Trivic, G. Wohlfahrt, J. Kandrac, D. Pericin, “Glucose oxidase from Aspergillus niger: the mechanism of action with molecular oxygen, quinones, and one-electron acceptors,” Int. J. Biochem. Cell Biol., 37 ( 4), 731-750, 2005.
  • N. Kurnaz Yetim, E. Hasanoğlu Özkan, B. Danış, H. Tümtürk, N. Sarı, “Research on the Repeated Use of Novel Ferrocene-Tagged Nanomaterial for Determination of Glucose,” Int. J. Polym. Mater. Polym. Biomater., 64, 888-893, 2015.
  • N. Kurnaz Yetim, E. Hasanoğlu Özkan, N. Sarı, “Immobilization of GOx on Trp / Trp-Fc Functionalized Nanospheres: Improved of Reusability and Stability,” Gazi Univ. J. Sci., 30 (4), 114-122, 2017.
  • A. H. Memon, R. Ding, Q. Yuan, H. Liang, Y. Wei, “Coordination of GMP ligand with Cu to enhance the multiple enzymesstability and substrate specificity by co-immobilization process,” Biochem. Eng. J.,136, 102-108, 2018.
  • P. Zhou, F. Li, W. Dong, K. Huang, Y. Chen, C. Wei, B. Ling, M. Li, “Covalent Immobilization of Glucose Oxidase onto Electrosynthesized Nanocomposite with PEDOT Derivative for Amperometric Glucose Biosensing,” Int. J. Electrochem. Sci.,13, 5294-5310, 2018.
  • H. Azak, H. B. Yildiz, B. Bezgin Carbas, “Synthesis and characterization of a new poly(dithieno (3,2-b:20, 30-d) pyrrole) derivative conjugated polymer: Its electrochromic and biosensing applications,” Polym., 134, 44-52, 2018.
  • C. Hou, D. Zhao, Y. Wang, S. Zhang, S. Li, “Preparation of magnetic Fe3O4/PPy@ZIF-8 nanocomposite for glucose oxidase immobilization and used as glucose electrochemical biosensor,” J. Electroanal. Chem., 822, 50-56, 2018.
  • S. Dönmez, F. Arslan, N. Sarı, N. Kurnaz Yetim, H. Arslan, “Preparation of carbon paste electrodes including poly(styrene) attached glycine-Pt(IV) for amperometric detection of glucose,” Biosens. Bioelectron., 54, 146-150, 2014.
  • J. Shim, G. Y. Kim, S. H. Moon, “Covalent co-immobilization of glucose oxidase and ferrocenedicarboxylic acid for an enzymatic biofuel cell,” J. Electroanal. Chem., 653, 14-20, 2011.
  • R. Ahmad, M. Sardar, “Enzyme Immobilization: An Overview on nanoparticles as immobilization matrix,” Biochem Anal. Biochem., 4, 1-8, 2015.
  • E. Çınar, S. Ercan, N. Güleşçi, “Lewatit Partikülüne Üreaz Enziminin İmmobilizasyonu,” Batman Univ. J. Life Sci., 7, 124-136, 2017.
  • N. Özdem, E. Hasanoğlu Özkan, N. Sarı, F. Arslan, H. Tümtürk, “Investigation of glucose oxidase enzyme as biocatalysts, new nanospheres attached azomethine,” Macromol. Res., 22 (12), 1282-1287, 2017.

Polimerik Nanoküreler Üzerine Enzim İmmobilizasyonu ve Optimizasyonu

Yıl 2019, Cilt: 14 Sayı: 1, 97 - 104, 31.05.2019
https://doi.org/10.29233/sdufeffd.479246

Öz

Enzimler, pek çok
endüstride kullanılan, maliyetleri oldukça yüksek olan önemli materyallerdir.
Katalitik tepkimelerde enzimler için şartlar uygun olsa bile yüksek sıcaklık,
yüksek veya düşük pH ve organik/inorganik çözücülerin varlığında enzimler aktifliklerini
kolaylıkla kaybedebilirler. İmmobilizasyon işlemi ile birlikte, enzimlerin
ekonomik açıdan kullanılabilirliğini sağlamak mümkündür. Bu amaçla,
(aminometil) polistiren (APS) kaynaklı nanokürelere glukoz oksidaz (GOx) enzimi
immobilize edildi ve optimizasyon parametreleri (pH, sıcaklık, tekrar kullanım
sayısı, depolama kararlılığı, substrat derişimi) belirlendi. Serbest ve
immobilize enzim için Michaelis-Menten eşitliği ile Km
ve Vmak değerleri hesaplandı. Özellikle Pt(IV) iyonu içeren
immobilize enzimin substratına karşı yüksek ilgi gösterdiği tespit edildi. 12
ay sonunda immobilize enzimlerin yaklaşık % 85 aktifliklerini korudukları
görüldü.

Kaynakça

  • N. Kurnaz Yetim, “Ferrosen içeren yeni Pt(II), Pt(IV) dendrimerlerinin sentezi ve immobilizasyonda kullanımı,” Doktora Tezi, Fen Bil. Enst., Gazi Üni., Ankara, Türkiye, 2016.
  • A. Wiseman, “Handbook of Enzymes Biotechnology (Book style with paper title and editor),” in The application of enzymes in industry, 2nd ed., E. Horwood, Ed. Chichester, UK, 1987, pp. 274-373.
  • E. Hasanoğlu Özkan, N. Kurnaz Yetim, D. Nartop, N. Sarı, “Influence of load on the recycling stability of nanospheres attached platinium ion for determination of glucose,” J. Ind. Eng. Chem., 25, 180-185, 2015.
  • S. Dönmez, F. Arslan, N. Sarı, E. Hasanoğlu Özkan, H. Arslan, “Glucose biosensor based on immobilization of glucose oxidase on a carbon paste electrode modified with microspheres attached L-glycine,” Biotechnol. Appl. Biochem., 64 (5), 745-753, 2017.
  • X. Wang, Y. Zhang, C. E. Banks, Q. Chen and X. Ji, “Non-enzymatic amperometric glucose biosensor based on nickel hexacyanoferrate nanoparticle film modified electrodes,” Colloids Surf. B, 78, 363-366, 2010.
  • H. Öğütcü, N. Kurnaz Yetim, E. Hasanoğlu Özkan, O. Eren, G. Kaya, N. Sarı, and A. Dişli, “Nanospheres Caped Pt(II) and Pt (IV): Synthesis and Evaluation as Antimicrobial and Antifungal Agent,” Pol. J. Chem., 19, 74-80, 2017.
  • P. Trinder, “Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor,” Ann. Clin. Biochem., 6, 24-27, 1969.
  • V. Leskovac, S. Trivic, G. Wohlfahrt, J. Kandrac, D. Pericin, “Glucose oxidase from Aspergillus niger: the mechanism of action with molecular oxygen, quinones, and one-electron acceptors,” Int. J. Biochem. Cell Biol., 37 ( 4), 731-750, 2005.
  • N. Kurnaz Yetim, E. Hasanoğlu Özkan, B. Danış, H. Tümtürk, N. Sarı, “Research on the Repeated Use of Novel Ferrocene-Tagged Nanomaterial for Determination of Glucose,” Int. J. Polym. Mater. Polym. Biomater., 64, 888-893, 2015.
  • N. Kurnaz Yetim, E. Hasanoğlu Özkan, N. Sarı, “Immobilization of GOx on Trp / Trp-Fc Functionalized Nanospheres: Improved of Reusability and Stability,” Gazi Univ. J. Sci., 30 (4), 114-122, 2017.
  • A. H. Memon, R. Ding, Q. Yuan, H. Liang, Y. Wei, “Coordination of GMP ligand with Cu to enhance the multiple enzymesstability and substrate specificity by co-immobilization process,” Biochem. Eng. J.,136, 102-108, 2018.
  • P. Zhou, F. Li, W. Dong, K. Huang, Y. Chen, C. Wei, B. Ling, M. Li, “Covalent Immobilization of Glucose Oxidase onto Electrosynthesized Nanocomposite with PEDOT Derivative for Amperometric Glucose Biosensing,” Int. J. Electrochem. Sci.,13, 5294-5310, 2018.
  • H. Azak, H. B. Yildiz, B. Bezgin Carbas, “Synthesis and characterization of a new poly(dithieno (3,2-b:20, 30-d) pyrrole) derivative conjugated polymer: Its electrochromic and biosensing applications,” Polym., 134, 44-52, 2018.
  • C. Hou, D. Zhao, Y. Wang, S. Zhang, S. Li, “Preparation of magnetic Fe3O4/PPy@ZIF-8 nanocomposite for glucose oxidase immobilization and used as glucose electrochemical biosensor,” J. Electroanal. Chem., 822, 50-56, 2018.
  • S. Dönmez, F. Arslan, N. Sarı, N. Kurnaz Yetim, H. Arslan, “Preparation of carbon paste electrodes including poly(styrene) attached glycine-Pt(IV) for amperometric detection of glucose,” Biosens. Bioelectron., 54, 146-150, 2014.
  • J. Shim, G. Y. Kim, S. H. Moon, “Covalent co-immobilization of glucose oxidase and ferrocenedicarboxylic acid for an enzymatic biofuel cell,” J. Electroanal. Chem., 653, 14-20, 2011.
  • R. Ahmad, M. Sardar, “Enzyme Immobilization: An Overview on nanoparticles as immobilization matrix,” Biochem Anal. Biochem., 4, 1-8, 2015.
  • E. Çınar, S. Ercan, N. Güleşçi, “Lewatit Partikülüne Üreaz Enziminin İmmobilizasyonu,” Batman Univ. J. Life Sci., 7, 124-136, 2017.
  • N. Özdem, E. Hasanoğlu Özkan, N. Sarı, F. Arslan, H. Tümtürk, “Investigation of glucose oxidase enzyme as biocatalysts, new nanospheres attached azomethine,” Macromol. Res., 22 (12), 1282-1287, 2017.
Toplam 19 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Kimya Mühendisliği
Bölüm Makaleler
Yazarlar

Nurdan Kurnaz Yetim 0000-0001-6227-0346

Elvan Hasanoğlu Özkan 0000-0001-7338-4015

Nurşen Sarı 0000-0002-4764-7747

Yayımlanma Tarihi 31 Mayıs 2019
Yayımlandığı Sayı Yıl 2019 Cilt: 14 Sayı: 1

Kaynak Göster

IEEE N. Kurnaz Yetim, E. Hasanoğlu Özkan, ve N. Sarı, “Polimerik Nanoküreler Üzerine Enzim İmmobilizasyonu ve Optimizasyonu”, Süleyman Demirel University Faculty of Arts and Science Journal of Science, c. 14, sy. 1, ss. 97–104, 2019, doi: 10.29233/sdufeffd.479246.