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Polyphenol Oxidase from Coleus forskohlii: Purification, Characterization, and Immobilization Onto Alginate/ZnO Nanocomposite Materials

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Abstract

Herein, polyphenol oxidase (PPO) was purified from Coleus forskohlii via a three-step process involving precipitation by (NH4)2SO4, ion exchange chromatography and gel filtration. PPO was purified 15-fold with a total yield of 31% and a specific activity of 3168 U/mg, and the observed molecular weight was 42 kDa. To improve the reusability and stability of the purified enzyme, calcium alginate incorporated with zinc oxide nanoparticles (Ca-ALG-ZnO NPs) was successfully prepared and characterized through scanning electron microscopy and Fourier transform infrared spectroscopy and utilized as a support for PPO immobilization. The immobilization yield and enzyme activity reached 83% and 3950 U/g support, respectively. The results showed that immobilized PPO could be reused ten times while retaining 69% of its original activity. The purified and immobilized PPO exhibited an optimal pH of 7.0 and 7.5 and an optimal temperature of 40 °C and 50 °C, respectively. The purified and immobilized PPO showed Vmax values of 255.75 and 251.89 Units/ml, and Km values of 4.99 and 3.12 mM, respectively. As a result, this work provides an opportunity to understand and manage the influence of the coleus forskohlii PPO enzyme in key immobilization, industrial. Additionally, the results indicated that the Ca-ALG-ZnO NPs used as supports had a significant effect on the enzyme activity and stability of immobilized PPO.

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References

  1. Das R, Talat M, Srivastava ON, Kayastha AM (2018) Covalent immobilization of peanut β-amylase for producing industrial nano-biocatalysts: A comparative study of kinetics, stability and reusability of the immobilized enzyme. Food chem 245:488–499

    Article  CAS  PubMed  Google Scholar 

  2. Singh K, Srivastava G, Talat M, Srivastava ON, Kayastha AM (2015) α-Amylase immobilization onto functionalized graphene nanosheets as scaffolds: Its characterization, kinetics and potential applications in starch based industries. Biochem Biophys Rep 3:18–25

    PubMed  PubMed Central  Google Scholar 

  3. Das R, Mishra H, Srivastava A, Kayastha AM (2017) Covalent immobilization of β-amylase onto functionalized molybdenum sulfide nanosheets, its kinetics and stability studies: A gateway to boost enzyme application. Chem Eng J 328:215–227

    Article  CAS  Google Scholar 

  4. Gan J, Bagheri AR, Aramesh N, Gul I, Franco M, Almulaiky YQ, Bilal M (2021) Covalent organic frameworks as emerging host platforms for enzyme immobilization and robust biocatalysis–A review. Int J Biol Macromol 167:502–515

    Article  CAS  PubMed  Google Scholar 

  5. Mohamed SA, Al-Harbi MH, Almulaiky YQ, Ibrahim IH, El-Shishtawy RM (2017) Immobilization of horseradish peroxidase on Fe3O4 magnetic nanoparticles. Electron J Biotechnol 27:84–90

    Article  CAS  Google Scholar 

  6. Almulaiky YQ, Aqlan FM, Aldhahri M, Baeshen M, Khan TJ, Khan KA, Alayafi AA (2018) α-Amylase immobilization on amidoximated acrylic microfibres activated by cyanuric chloride. R. Soc. Open sci. 5:172164

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Attard GS, Bartlett PN, Coleman NR, Elliott JM, Owen JR, Wang JH (1997) Mesoporous platinum films from lyotropic liquid crystalline phases. Science 278:838–840

    Article  CAS  Google Scholar 

  8. Sun J, Zhang H, Tian R, Ma D, Bao X, Su DS, Zou H (2006) Ultrafast enzyme immobilization over large-pore nanoscale mesoporous silica particles. Chem Commun 12:1322–1324

    Article  Google Scholar 

  9. Abdel-Mageed HM, El-Laithy HM, Mahran LG, Fahmy AS, Mäder K, Mohamed SA (2012) Development of novel flexible sugar ester vesicles as carrier systems for the antioxidant enzyme catalase for wound healing applications. Process Beachem 47:1155–1162

    Article  CAS  Google Scholar 

  10. Strand BL, Morch YA, Skjak-Braek G (2000) Alginate as immobilization matrix for cells. Minerva Biotecnologica 12:223

    Google Scholar 

  11. Kamaldeep DK, Kashyap KD (2012) Optimization of zinc oxide nanoparticles synthesis to fabricate glucose oxidase sensor. Adv Appl Sci Res 3:3081–3088

    CAS  Google Scholar 

  12. Alfadul SM, Elneshwy AA (2010) Use of nanotechnology in food processing, packaging and safety review. Afr J Food Agric Nutr Dev 10:2719–2739

    CAS  Google Scholar 

  13. Gong Z, Li D, Liu C, Cheng A, Wang W (2015) Partial purification and characterization of polyphenol oxidase and peroxidase from chestnut kernel. LWT-Food Sci Technol 60:1095–1099

    Article  CAS  Google Scholar 

  14. Falguera V, Sánchez-Riaño A, Quintero-Cerón J, Rivera-Barrero C, Méndez- Arteaga J, Ibarz A (2012) Characterization of polyphenol oxidase activity in juices from 12 underutilized tropical fruits with high agroindustrial potential. Food Bioproc. Technol. 5:2921–2927

    Article  CAS  Google Scholar 

  15. Al-Najadaa AR, Mohamed SA (2014) Changes of antioxidant capacity and oxidoreductases of Saudi date cultivars (Phoenix dactylifera L.) during storage. Sci Hortic 170:275–280

    Article  Google Scholar 

  16. Queiroz C, Mendes Lopes ML, Fialho E, Valente-Mesquita VL (2008) Polyphenol oxidase: characteristics and mechanisms of browning control. Food Rev Int 24:361–375

    Article  CAS  Google Scholar 

  17. Aitken MD, Massey IJ, Chen T, Heck PE (1994) Characterization of reaction products from the enzyme catalyzed oxidation of phenolic pollutants. Water Res 28:1879–1889

    Article  CAS  Google Scholar 

  18. Edalli VA, Kamanavalli CM (2010) Removal of phenolic compounds by mushroom polyphenol oxidase from Pleurotus species. Bioscan 4:89–92

    CAS  Google Scholar 

  19. Mukherjee S, Basak B, Bhunia B, Dey A, Mondal B (2013) Potential use of polyphenol oxidases (PPO) in the bioremediation of phenolic contaminants containing industrial wastewater. Rev Environ Sci Biotechnol 12:61–73

    Article  CAS  Google Scholar 

  20. Guimarães CO, França AB, Samanamud GRL, Baston EP, Lofrano RCZ, Loures CCA, Naves FL (2019) Optimization of treating phenol from wastewater through the TiO 2-catalyzed advanced oxidation process and response surface methodology. Environ Monit Assess 191:349

    Article  Google Scholar 

  21. Shukla PK, Misra A, Kumar M, Jaichand SK, Akhtar J, Srivastava S, Agrawal PK, Singh AK (2018) Rawat, Simultaneous quantification of forskolin and iso-forskolin in Coleus forskohlii (Wild.) Briq. and identification of elite chemotype, collected from eastern ghats (India). Pharmacogn Mag 13:881–885

    Google Scholar 

  22. Almulaiky YQ, Kuerban A, Aqlan F, Alzahrani SA, Baeshen MN, Afifi M, Alkhaled M (2017) In vitro antiglycation, antioxidant properties of coleus forskohlii “Balady” leaves and stem and their antioxidant enzyme activities. Annu Res Rev 19:1–11

    Google Scholar 

  23. Ding X, Staudinger JL (2005) Induction of drug metabolism by forskolin: the role of the pregnane X receptor and the protein kinase a signal transduction pathway. J Pharmacol Exp Ther 312:849–856

    Article  CAS  PubMed  Google Scholar 

  24. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  25. Dahham SS, Al-Rawi SS, Ibrahim AH, Majid ASA, Majid SAM (2018) Antioxidant, anticancer, apoptosis propertiesand chemical composition of black truffle Terfezia claveryi. Saudi J. Biol. Sci. 25:1524–1534

    Article  CAS  PubMed  Google Scholar 

  26. Benaceur F, Chaibi R, Berrabah F, Neifar A, Leboukh M, Benaceur K, Gargouri A (2020) Purification and characterization of latent polyphenol oxidase from truffles (Terfezia arenaria). Int J Biol Macromol 145:885–893

    Article  CAS  PubMed  Google Scholar 

  27. Almulaiky YQ, Al-Harbi SA (2019) A novel peroxidase from Arabian balsam (Commiphora gileadensis) stems: Its purification, characterization and immobilization on a carboxymethylcellulose/Fe3O4 magnetic hybrid material. Int J Biol Macromol 133:767–774

    Article  CAS  PubMed  Google Scholar 

  28. Eddehech A, Zied Z, Aloui F, Smichi N, Noiriel A, Abousalham A, Gargouri Y (2019) Production, purification, and biochemical characterization of a thermoactive, alkaline lipase from a newly isolated Serratia sp. W3 Tunisian strain. Int J Biol Macromol 123:792–800

    Article  CAS  PubMed  Google Scholar 

  29. Govindarajan RK, Krishnamurthy M, Neelamegam R, Shyu DJ, Muthukalingan K, Nagarajan K (2018) Purification, structural characterization and biotechnological potential of tannase enzyme produced by Enterobacter cloacae strain 41. Process Biochem 77:37–47

    Google Scholar 

  30. Al-Harbi SA, Almulaiky YQ (2020) Purification and biochemical characterization of Arabian balsam α-amylase and enhancing the retention and reusability via encapsulation onto calcium alginate/Fe2O3 nanocomposite beads Int. J Biol Macromol 160:944–952

    Article  CAS  Google Scholar 

  31. Almulaiky YQ (2020) Peroxidase from coleus forskohlii: purification and biochemical characterization. Int J Nutr 5:9–20

    Article  CAS  Google Scholar 

  32. Almulaiky YQ, Al-Harbi SA (2021) Preparation of a calcium alginate-coated polypyrrole/silver nanocomposite for site-specific immobilization of polygalacturonase with high reusability and enhanced stability. Catal Letters. https://doi.org/10.1007/s10562-021-03631-7

    Article  Google Scholar 

  33. Aksoy MA (2020) new insight into purification of polyphenol oxidase and inhibition effect of curcumin and quercetin on potato polyphenol oxidase. Protein Expr Purif 171:1–6

    Article  Google Scholar 

  34. Öztürk C, Aksoy M, Küfrevioğlu OI (2020) Purification of tea leaf (Camellia sinensis) polyphenol oxidase by using affinity chromatography and investigation of its kinetic properties. J Food Meas Charact 14:31–38

    Article  Google Scholar 

  35. Vishwasrao C, Ananthanarayan L (2018) Partial purification and characterization of the quality deteriorating enzymes from Indian pink guava (Psidium guajava L.), var. Lalit. J. Food Sci. Technol. 55:3281–3291

    Article  CAS  PubMed  Google Scholar 

  36. Teng J, Gong Z, Deng Y, Chen L, Li Q, Shao Y, Xiao W (2017) Purification, characterization, and enzymatic synthesis of theaflavins of polyphenol oxidase isozymes from tea leaf (Camellia sinensis). LWT 84:263–270

    Article  CAS  Google Scholar 

  37. Li S, Zhong L, Wang H, Li J, Cheng H, Ma Q (2021) Process optimization of polyphenol oxidase immobilization: Isotherm, kinetic, thermodynamic and removal of phenolic compounds. Int J Biol Macromol 185:792–803

    Article  CAS  PubMed  Google Scholar 

  38. Zhang F, Zheng B, Zhang J, Huang X, Liu H, Guo S, Zhang J (2010) Horseradish peroxidase immobilized on graphene oxide: physical properties and applications in phenolic compound removal. J Phys Chem 114:8469–8473

    CAS  Google Scholar 

  39. El-Shishtawy RM, Ahmed NS, Almulaiky YQ (2021) Immobilization of catalase on chitosan/zno and chitosan/ZnO/Fe2O3 nanocomposites: A comparative study. Catalyst 11:820

    Article  CAS  Google Scholar 

  40. Wang H, Li S, Li J, Zhong L, Cheng H, Ma Q (2020) Immobilized polyphenol oxidase: Preparation, optimization and oxidation of phenolic compounds. Int J Biol Macromol 160:233–244

    Article  CAS  PubMed  Google Scholar 

  41. Lončar N, Vujčić Z (2011) Tentacle carrier for immobilization of potato phenoloxidase and its application for halogenophenols removal from aqueous solutions. J Hazar Mater 196:73–78

    Article  Google Scholar 

  42. Shao J, Huang LL, Yang YM (2009) Immobilization of polyphenol oxidase on alginate–SiO2 hybrid gel: stability and preliminary applications in the removal of aqueous phenol. J Chem Technol Biotechnol 84:633–635

    Article  CAS  Google Scholar 

  43. Al-Najada AR, Almulaiky YQ, Aldhahri M, El-Shishtawy RM, Mohamed SA, Baeshen M, Al-Harbi SA (2019) Immobilisation of α-amylase on activated amidrazone acrylic fabric: a new approach for the enhancement of enzyme stability and reusability. Sci Rep 9:1–9

    Article  CAS  Google Scholar 

  44. Almulaiky YQ, Khalil NM, El-Shishtawy RM, Altalhi T, Algamal Y, Aldhahri M, Mohammed MM (2021) Hydroxyapatite-decorated ZrO2 for α-amylase immobilization: Toward the enhancement of enzyme stability and reusability. Int J Biol Macromol 167:299–308

    Article  CAS  PubMed  Google Scholar 

  45. Shi C, Dai Y, Liu Q, Xie Y, Xu X (2003) The FT-IR spectrometric analysis of the changes of polyphenol oxidase II secondary structure. J Mol Struct 644:139–144

    Article  CAS  Google Scholar 

  46. Alici EH, Arabaci G (2016) Purification of polyphenol oxidase from borage (Trachystemo norientalis L.) by using three-phase partitioning and investigation of kinetic properties. Int J Biol Macromol 93:1051–1056

    Article  CAS  PubMed  Google Scholar 

  47. Derardja AE, Pretzler M, Kampatsikas I, Barkat M, Rompel A (2017) Purification and characterization of latent polyphenol oxidase from apricot (Prunus armeniaca L.). J Agric Food Chem 65:8203–8212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Atiroğlu V, Atiroğlu A, Özacar M (2021) Immobilization of α-amylase enzyme on a protein@ metal–organic framework nanocomposite: A new strategy to develop the reusability and stability of the enzyme. Food Chem 349:129127

    Article  PubMed  Google Scholar 

  49. Akgol S, Kacar Y, Denizli A, Arica MY (2001) Hydrolysis of sucrose by invertase immobilized onto novel magnetic polyvinyl alcohol microspheres. Food Chem 74:281–288

    Article  CAS  Google Scholar 

  50. Razzaque MA, Saud ZA, Absar N (2000) Purification and characterization of polyphenoloxidase from guava infected with fruit-rot disease. Pak J Biol Sci 3:407–410

    Article  Google Scholar 

  51. Jia G, Baogang W, Xiaoyuan F, Haoru T, Wensheng L, Kaichun Z (2011) Partial properties of polyphenol oxidase in sour cherry (Prunus cerasus L.) Pulp. World J. Agric. Sci. 7:444–449

    Google Scholar 

  52. Cao SL, Li XH, Lou WY, Zong MH (2014) Preparation of a novel magnetic cellulose nanocrystal and its efficient use for enzyme immobilization. J Mater Chem B 2:5522–5530

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The work was funded by the University of Jeddah, Saudi Arabia, under Grant No. (UJ-20- 076-DR). The authors, therefore, acknowledge with thanks the university technical and support.

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Authors and Affiliations

  1. Department of Chemistry, College of Science and Arts at Khulis, University of Jeddah, Jeddah, Saudi Arabia

    Yaaser Q. Almulaiky

  2. Chemistry Department, Faculty of Applied Science, Taiz University, Taiz, Yemen

    Yaaser Q. Almulaiky

  3. Department of Biology, College of Science, University of Jeddah, Jeddah, Saudi Arabia

    Omar Almaghrabi

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Almulaiky, Y.Q., Almaghrabi, O. Polyphenol Oxidase from Coleus forskohlii: Purification, Characterization, and Immobilization Onto Alginate/ZnO Nanocomposite Materials. Catal Lett 152, 3089–3099 (2022). https://doi.org/10.1007/s10562-022-03916-5

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