Flowerlike hybrid horseradish peroxidase nanobiocatalyst for the polymerization of guaiacol

In this study, the catalytic activity and stability of flowerlike hybrid horseradish peroxidase (HRP) nanobiocatalyst (HRPCu2+) obtained from $Cu^{2+}$ ions and HRP enzyme in the polymerization reaction of guaiacol were analyzed. We demonstrated that HRP-Cu2+ and hydrogen peroxide $(H_2O_2)$ initiator showed significantly increased catalytic activity and stability on the polymerization of guaiacol compared to that of free HRP enzyme. Poly(guaiacol) was observed with quite high yields (88%) and molecular weights (38,000 g/mol) under pH 7.4 phosphate-buffered saline (PBS) conditions at 60 °C with 5 weight% of $HRP-Cu^{2+}$ loading. $HRP-Cu^{2+}$ also shows very high thermal stability and works even at 70 °C reaction temperature; free HRP enzyme denatures at that temperature. Furthermore, $HRP-Cu^{2+}$ provided considerable repeated use and showed some degree of catalytic activity, even after the fourth recycle, in the polymerization of guaiacol.

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1. Horton HR, Moran LA, Ochs RS. Principles of Biochemistry. New Jersey, USA: Prentice-Hall International Inc., 1996.
2. Ansari SA, Husain Q. Potential applications of enzymes immobilized on/in nano materials: a review. Biotechnology Advances 2012; 30 (3): 512-523. doi: 10.1016/j.biotechadv.2011.09.005
3. Tran DN, Balkus KJ. Perspective of recent progress in immobilization of enzymes. ACS Catalysis2011; 1 (8): 956-968. doi: 10.1021/ cs200124a
4. Jia F, Narasimhan B, Mallapragada S. Materials-based strategies for multi-enzyme immobilization and co-localization: a Review. Biotechnology and Bioengineering 2014; 111 (2): 209-222. doi: 10.1002/bit.25136
5. DiCosimo R, McAuliffe J, Pouloseb AJ, Bohlmann G. Industrial use of immobilized enzymes. Chemical Society Reviews 2013; 42 (15): 6437-6474. doi: 10.1039/c3cs35506c
6. Ahmad R, Sardar M. Enzyme Immobilization: an overview on nanoparticles as immobilization matrix. Biochemistry and Analytical Biochemistry 2015; 4 (2): 178-186. doi:10.4172/2161-1009.1000178
7. Mateo C, Palomo JM, Fernandez-Lorente G, Guisan JM, Fernandez-Lafuente R. Improvement of enzyme activity: stability and selectivity via immobilization techniques. Enzyme and Microbial Technology 2007; 40 (6): 1451-1463. doi: 10.1016/j.enzmictec.2007.01.018
8. Hollmann F, Arends IWCE. Enzyme initiated radical polymerizations. Polymers 2012; 4 (1): 759-793. doi: 10.3390/polym4010759
9. Dordick JS, Marletta MA, Klibanov AM. Polymerization of phenols catalyzed by peroxidase in nonaqueous media. Biotechnology and Bioengineering 1987; 30 (1): 31-36. doi: 10.1002/bit.260300106
10. Tonami H, Uyama H, Kobayashi S, Rettig K, Ritter H. Chemoenzymatic synthesis of a poly(hydroquinone). Macromolecular Chemistry and Physics 1999; 200 (9): 1998-2002.doi:10.1002/(SICI)1521-3935(19990901)200:9<1998::AID-MACP1998>3.0.CO;2-6
11. Goretzki C, Ritter H. Enzymatic oxidative polymerization of aminochalcones by use of horseradish peroxidase. Macromolecular Chemistry and Physics 1998; 199 (6): 1019-1024. doi: 10.1002/(SICI)1521-3935(19980601)199:6<1019::AID-MACP1019>3.0.CO;2-5
12. Uyama H, Kurioka H, Sugihara J, Komatsu I, Kobayashi S. Oxidative polymerization of p-alkylphenols catalyzed by horseradish peroxidase. Journal of Polymer Science Part A: Polymer Chemistry 1997; 35 (8): 1453-1459. doi: 10.1002/(SICI)1099-0518(199706)35:8<1453::AIDPOLA14>3.0.CO;2-6
13. Uyama H, Kobayashi S. Enzyme-catalyzed polymerization to functional polymers. Journal of Molecular Catalysis B: Enzymatic 2002; 19- 20: 117-127. doi: 10.1016/S1381-1177(02)00158-3
14. Uyama H, Lohavisavapanich C, Ikedia R, Kobayashi S. Chemoselective polymerization of a phenol derivative having a methacryl group by peroxidase catalyst. Macromolecules 1998; 31 (2): 554-556. doi: 10.1021/ma971510p
15. Kocak A, Kumbul A, Gokturk E, Sahmetlioglu E. Synthesis and characterization of imine-functionalized polyphenol via enzymatic oxidative polycondensation of a bisphenol derivative. Polymer Bulletin 2016; 73 (1): 163-177. doi: 10.1007/s00289-015-1478-1
16. Isci I, Gokturk E, Turac E, Sahmetlioglu E. Chemoenzymatic polymerization of hydrazone functionalized phenol. Polymer Science Series B 2016; 58 (4): 411-420. doi: 10.1134/S1560090416040047
17. Zhang Y, Ge J, Liu Z. Enhanced activity of immobilized or chemically modified enzymes. ACS Catalysis 2015; 5: 4503-4513. doi: 10.1021/ acscatal.5b00996
18. Kragl U. Immobilized Enzymes and Membrane Reactor.In: Godfrey T, West S (editors). Industrial Enzymology.London, UK: Macmillan Press, 1996, pp. 275-282.
19. Hanefeld U, Cao L, Magner E. Enzyme immobilisation: fundamentals and application. Chemical Society Reviews 2013; 42: 15. doi: 10.1039/c3cs90042h
20. Gupta MN, Mattiasson B. Unique applications of immobilized proteins in bioanalytical systems. Methods of BiochemicalAnalysis2006; 36: 1-34. doi: 10.1002/9780470110577.ch1
21. Altinkaynak C, Tavlasoglu S, Ozdemir N, Ocsoy I. A new generation approach in enzyme immobilization: Organic-inorganic hybrid nanoflowers with enhanced catalytic activity and stability. Enzyme and Microbial Technology 2016; 93-94: 105-112. doi: 10.1016/j. enzmictec.2016.06.011
22. Li Y, Wu H, Su Z. Enzyme-based hybrid nanoflowers with high performances for biocatalytic, biomedical, and environmental applications. Coordination Chemistry Reviews, 2020; 416: 213342. doi: 10.1016/j.ccr.2020.213342
23. Zhang X, Gong C, Akakuru OU, Su Z, Wu A, Wei G. The design and biomedical applications of self-assembled two-dimensional organic biomaterials. Chemical Society Reviews, 2019; 48: 5564-5595. doi: 10.1039/c8cs01003j
24. Xiong Q, Zhang X, Wei W, Wei G, Su Z. Enzyme-mediated reversible deactivation radical polymerization for functional materials: principles, synthesis, and applications. Polymer Chemistry, 2020; 11: 1673-1690. doi: 10.1039/D0PY00136H
25. Ge J, Lei J, Zare RN. Protein–inorganic hybrid nanoflowers. Nature Nanotechnology 2012; 7: 428-432. doi: 10.1038/nnano.2012.80
26. Ocsoy I, Dogru E, Usta S. A new generation of flowerlike horseradish peroxides as a nanobiocatalyst for superior enzymatic activity. Enzyme and Microbial Technology 2015; 75-76: 25-29. doi: 10.1016/j.enzmictec.2015.04.010
27. Somturk B, Hancer M, Ocsoy I, Ozdemir N. Synthesis of copper ion incorporated horseradish peroxidase-based hybrid nanoflowers for enhanced catalytic activity and stability. Dalton Transactions 2015; 44: 13845-13852. doi: 10.1039/C5DT01250C
28. Lancefield CS, Weckhuysen BM, Bruijnincx PCA. Catalytic Conversion of Lignin-derived Aromatic Compounds into Chemicals. In: Beckham GT (editor). Lignin Valirization: Emerging Approaches. London, UK: Royal Society of Chemistry, 2018, pp. 159-198.
29. Topal Y, Tapan S, Gokturk E, Sahmetlioglu E. Horseradish peroxidase-catalyzed polymerization of ortho-imino-phenol: synthesis, characterization, thermal stabilityand electrochemical properties. Journal of Saudi Chemical Society 2017; 21 (6): 731-740. doi: 10.1016/j. jscs.2017.03.006
30. Yildirim P, Gokturk E, Turac E, Demir HO, Sahmetlioglu E. Chemoenzymatic polycondensation of para-benzylamino phenol. Chemical Papers 2016; 70 (5): 610-619. doi: 10.1515/chempap-2015-0242
31. Nanayakkara S, Zhao Z, Patti AF, He L, Saito K. Immobilized Horseradish Peroxidase (I-HRP) as Biocatalyst for Oxidative Polymerization of 2,6-Dimethylphenol. ACS Sustainable Chemistry and Engineering2014; 2 (8): 1947-1950. doi: 10.1021/sc500392k
32. Kumbul A, Gokturk E, Turac E, Sahmetlioglu E. Enzymatic oxidative polymerization of para-imine functionalized phenol catalyzed by horseradish peroxidase. Polymers for Advanced Technologies 2015; 26 (9): 1123-1129. doi: 10.1002/pat.3544
33. Kumbul A, Gokturk E, Sahmetlioglu E. Synthesis, characterization, thermal stability and electrochemical properties of ortho-iminefunctionalized oligophenol via enzymatic oxidative polycondensation. Journal of Polymer Research 2016; 23 (52). doi: 10.1007/s10965- 016-0953-1
34. Gokturk E, Ocsoy I, Turac E, Sahmetlioglu E. Horseradish peroxidase-based hybrid nanoflowers with enhanced catalytical activities for polymerization reactions of phenol derivatives. Polymers for Advanced Technologies, 2020, 1-7. doi: 10.1002/pat.4956
35. Burrows A, Holman J, Parsons A, Pilling G, Price G. Chemistry3: Introducing Inorganic, Organic and Physical Chemistry. 3rd edition, Oxford, UK: Oxford university press, 2017.
36. Iwahara K, Honda Y, Watanabe T, KuwaharaM. Polymerization of guaiacol by lignin-degrading manganese peroxidase from Bjerkanderaadusta in aqueous organic solvents. Applied Microbiology and Biotechnology 2000; 54: 104-111. doi: 10.1007/s002530000340
37. Hwang S, Lee Y-W, Lee C-H, Ahn I-K. Manganese (III) Acetate-Catalyzed Synthesis of Polyguaiacol. Journal of Polymer Science: Part A: Polymer Chemistry 2008; 46: 6009-6015. doi: 10.1002/pola.22902