Journal of Sciences, Islamic Republic of Iran

Current Issue

By Issue

By Author

By Subject

Author Index

Keyword Index

About Journal

Aims and Scope

Editorial Board

Publication Ethics

Indexing and Abstracting

Related Links

FAQ

Peer Review Process

Journal Metrics

News

The Role of Cellulose Binding Heptapeptide in Immobilizing Glucose Oxidase to Cellulosic Surfaces

Document Type : Original Paper

Authors

1 1 Department of Molecular Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Islamic Republic of Iran 2 Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Islamic Republic of Iran

2 1 Department of Molecular Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Islamic Republic of Iran 3 Department of Systems Biotechnology, Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Islamic Republic of Iran

3 2 Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Islamic Republic of Iran

4 3 Department of Systems Biotechnology, Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Islamic Republic of Iran

5 1 Department of Molecular Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Islamic Republic of Iran

Abstract

This study aimed to produce a recombinant version of glucose oxidase enzyme and introduce an economical method for enzyme immobilization. The pGAPZαA expression vector was utilized, which has a continuous expression and does not need an inducer and medium replacement. Enzyme immobilization was accomplished by adding WHWTYYW heptapeptide, which tends to bind to cellulosic and nitrocellulose surfaces, to bind the enzyme to the above-mentioned surfaces. In this study, the glucose oxidase (GOX)-encoding gene from Aspergillus niger ATCC 9029 was amplified by the polymerase chain reaction (PCR) and cloned into the pGAPZαA vector, and pGAPZαA/GOX was obtained as a control construct. On the other hand, a primer containing the heptapeptide sequence was designed, followed by adding the heptapeptide by the PCR technique to the end of the carboxyl in the GOX-encoding gene in the pGAPZαA/GOX construct and obtaining the pGAPZαA/GOX–heptapeptide construct. Both constructs were integrated into the Pichia pastoris genome by electroporation, and enzymatic activity and protein expression of the recombinant enzyme from both constructs underwent investigation. The amount of GOX production in the pGAPZαA/GOX wild-type construct and pGAPZαA/GOX with heptapeptide was estimated at 0.234 and 0.135 g/l, respectively. The stability of both constructs was evaluated on different surfaces and then compared as well. It was revealed that the enzyme with WHWTYYW heptapeptide has an increased affinity to bind in cellulose and nitrocellulose surfaces compared to the wild-type enzyme, while demonstrating no affinity for binding to the polyvinylidene fluoride paper.

Keywords

References
  1. Morshed MN, Behary N, Bouazizi N, Guan J, Chen G, Nierstrasz V. Surface modification of polyester fabric using plasma-dendrimer for robust immobilization of glucose oxidase enzyme. Sci Rep. 2019;9(1):1-6.
  2. Varkani RA, Rafiee-Pour HA, Noormohammadi M. One step immobilization of glucose oxidase on TiO2 nanotubes towards glucose biosensing. Microchem. 2021;170:106712.
  3. Ning X, Zhang Y, Yuan T, Li Q, Tian J, Guan W, et al. Enhanced thermostability of glucose oxidase through computer-aided molecular design. Int J Mol Sci. 2018;19(2):425.
  4. Wong CM, Wong KH, Chen XD. Glucose oxidase: natural occurrence, function, properties and industrial applications. AMAB. 2008;78(6):927-38.
  5. Bankar SB, Bule MV, Singhal RS, Ananthanarayan L. Glucose oxidase—an overview. Biotechnol Adv. 2009;27(4):489-501.
  6. Ramachandran S, Fontanille P, Pandey A, Larroche C. Gluconic acid: properties, applications and microbial production. Food Technol Biotechnol. 2006 Apr 1;44(2).185-95.
  7. Babu VS, Kumar MA, Karanth NG, Thakur MS. Stabilization of immobilized glucose oxidase against thermal inactivation by silanization for biosensor applications. Biosens Bioelectron. 2004;19(10):1337-41.
  8. Tirkeş S, Toppare L, Alkan S, Bakir U, Önen A, Yağcı Y. Immobilization of glucose oxidase in polypyrrole/polytetrahydrofuran graft copolymers. Int J Biol Macromol. 2002;30(2):81-7.
  9. Tamerler C, Sarikaya M. Molecular biomimetics: genetic synthesis, assembly, and formation of materials using peptides. MRS Bull. 2008;33(5):504-12.
  10. So CR, Kulp III JL, Oren EE, Zareie H, Tamerler C, Evans JS, Sarikaya M. Molecular recognition and supramolecular self-assembly of a genetically engineered gold binding peptide on Au {111}. ACS Nano. 2009;3(6):1525-31.
  11. Willner I, Willner B. Biomolecule-based nanomaterials and nanostructures. Nano Lett. 2010;10(10):3805-15.
  12. Schulz C, Kittl R, Ludwig R, Gorton L. Direct electron transfer from the FAD cofactor of cellobiose dehydrogenase to electrodes. ACS Catal. 2016;6(2):555-63.
  13. Muguruma H, Iwasa H, Hidaka H, Hiratsuka A, Uzawa H. Mediatorless direct electron transfer between flavin adenine dinucleotide-dependent glucose dehydrogenase and single-walled carbon nanotubes. ACS Catal. 2017;7(1):725-34.
  14. Nguyen HH, Lee SH, Lee UJ, Fermin CD, Kim M. Immobilized enzymes in biosensor applications. Mater. 2019;12(1):121.
  15. Mohamad NR, Marzuki NH, Buang NA, Huyop F, Wahab RA. An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes. Biotechnol Biotechnol Equip. 2015;29(2):205-20.
  16. Balland V, Hureau C, Cusano AM, Liu Y, Tron T, Limoges B. Oriented immobilization of a fully active monolayer of histidine‐tagged recombinant laccase on modified gold electrodes. Chem. 2008;14(24):7186-92.
  17. Sassolas A, Blum LJ, Leca-Bouvier BD. Immobilization strategies to develop enzymatic biosensors. Biotechnol Adv. 2012;30(3):489-511.
  18. Vatsyayan P. Recent advances in the study of electrochemistry of redox proteins. Trend Bioelectroanal. 2016:223-62.
  19. Sarikaya M, Aksay IA. Biomimetics: design and processing of materials. In AIP series in Polym. Complex Mater. 1995:147-160.
  20. So CR, Hayamizu Y, Yazici H, Gresswell C, Khatayevich D, Tamerler C, Sarikaya M. Controlling self-assembly of engineered peptides on graphite by rational mutation. ACS Nano. 2012;6(2):1648-56.
  21. Kacar T, Zin MT, So C, Wilson B, Ma H, Gul‐Karaguler N, Jen AK, Sarikaya M, Tamerler C. Directed self‐immobilization of alkaline phosphatase on micro‐patterned substrates via genetically fused metal‐binding peptide. Biotechnol Bioeng. 2009;103(4):696-705.
  22. Hnilova M, Liu X, Yuca E, Jia C, Wilson B, Karatas AY, Gresswell C, Ohuchi F, Kitamura K, Tamerler C. Multifunctional protein-enabled patterning on arrayed ferroelectric materials. ACS Appl Mater Interfaces. 2012;4(4):1865-71.
  23. Hnilova M, Karaca BT, Park J, Jia C, Wilson BR, Sarikaya M, Tamerler C. Fabrication of hierarchical hybrid structures using bio‐enabled layer‐by‐layer self‐assembly. Biotechnol Bioeng. 2012;109(5):1120-30.
  24. Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 2000;24(1):45-66.
  25. Cregg JM, Tolstorukov I, Kusari A, Sunga J, Madden K, Chappell T. Expression in the yeast Pichia pastoris. Meth Enzymol. 2009;463:169-89.
  26. Abad S, Nahalka J, Winkler M, Bergler G, Speight R, Glieder A, Nidetzky B. High-level expression of Rhodotorula gracilis d-amino acid oxidase in Pichia pastoris. Biotechnol Lett. 2011;33(3):557-63.
  27. Briand L, Nespoulous C, Huet JC, Takahashi M, Pernollet JC. Ligand binding and physico‐chemical properties of ASP2, a recombinant odorant‐binding protein from honeybee (Apis mellifera L.). EJB. 2001;268(3):752-60.
  28. Briand L, Eloit C, Nespoulous C, Bézirard V, Huet JC, Henry C, Blon F, Trotier D, Pernollet JC. Evidence of an odorant-binding protein in the human olfactory mucus: location, structural characterization, and odorant-binding properties. Biochem. 2002;41(23):7241-52.
  29. Cereghino GP, Cereghino JL, Ilgen C, Cregg JM. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr Opin Biotechnol. 2002;13(4):329-32.
  30. Ahmad M, Hirz M, Pichler H, Schwab H. Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. AMAB. 2014;98(12):5301-17.
  31. Belyad F, Karkhanei AA, Raheb J. Expression, characterization and one step purification of heterologous glucose oxidase gene from Aspergillus niger ATCC 9029 in Pichia pastoris. EuPA Open Proteom. 2018; 19:1-5.
  32. Fasihi M, Rahpeyma SS, Karkhanei AA, Raheb J. Optimization of Synthetic Glucose Oxidase Gene Using A Recombinant Combination Strategy in Pichia Pastoris GS115. J Sci Islam Repub Iran. 2022;33(1):5-18.
  33. Gidijala L, Uthoff S, van Kampen SJ, Steinbüchel A, Verhaert R. Presence of protein production enhancers results in significantly higher methanol-induced protein production in Pichia pastoris. Microb. Cell Fact. 2018;17(1):1-1.
  34. Chen CC, Wu PH, Huang CT, Cheng KJ. A Pichia pastoris fermentation strategy for enhancing the heterologous expression of an Escherichia coli phytase. Enzyme Microb Technol. 2004;35(4):315-20.
  35. Vieira Gomes AM, Souza Carmo T, Silva Carvalho L, Mendonça Bahia F, Parachin NS. Comparison of yeasts as hosts for recombinant protein production. Microorganisms. 2018;6(2):38.
  36. Guo J, Catchmark JM, Mohamed MN, Benesi AJ, Tien M, Kao TH, et al. Identification and characterization of a cellulose binding heptapeptide revealed by phage display. Biomacromolecules. 2013;14(6):1795-805.
  37. Xu GY, Ong E, Gilkes NR, Kilburn DG, Muhandiram DR, Harris-Brandts M, et al. Solution structure of a cellulose-binding domain from Cellulomonas fimi by nuclear magnetic resonance spectroscopy. Biochemistry. 1995;34(21):6993-7009.
  38. Tormo J, Lamed R, Chirino AJ, Morag E, Bayer EA, Shoham Y, et al. Crystal structure of a bacterial family‐III cellulose‐binding domain: a general mechanism for attachment to cellulose. EMBO J. 1996;15(21):5739-51.
  39. Poveda A, Jiménez-Barbero J. NMR studies of carbohydrate–protein interactions in solution. Chem Soc Rev. 1998;27(2):133-44.
  40. Boraston AB, Bolam DN, Gilbert HJ, Davies GJ. Carbohydrate-binding modules: fine-tuning polysaccharide recognition. Biochem J. 2004;382(3):769-81.
  41. Khandare JJ, Minko T. Antibodies and peptides in cancer therapy. Crit Rev Ther Drug Carrier Syst. 2006;23(5).
  42. Linder M, Mattinen ML, Kontteli M, Lindeberg G, Ståhlberg J, Drakenberg T, et al. Identification of functionally important amino acids in the cellulose‐binding domain of Trichoderma reesei cellobiohydrolase I. Protein Sci. 1995;4(6):1056-64.
  43. Reinikainen T, Teleman O, Teeri TT. Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei. Proteins. 1995;22(4):392-403.
  44. Zhou K, Zhu Y, Yang X, Li C. Electrocatalytic oxidation of glucose by the glucose oxidase immobilized in graphene‐au‐nafion biocomposite. Electroanalysis. 2010;22(3):259-64.
  45.  

 
  • Sheldon RA. Enzyme immobilization: the quest for optimum performance. Adv Synth Catal. 2007;349(8‐9):1289-307.
  1. Serizawa T, Iida K, Matsuno H, Kurita K. Cellulose-binding heptapeptides identified by phage display methods. Chem Lett. 2007;36(8):988-9.
Journal of Sciences, Islamic Republic of Iran
Volume 33, Issue 3
September 2022
Pages 213-225
Files
Share
How to cite
Statistics