Vegetos
(An International Journal of Plant Research & Biotechnology)
(eISSN: 2229-4473)
Purification, biochemical properties and inhibition of PPO from breadfruit (Artocarpus altilis) by natural fruit juices
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Research Articles | Published: 04 July, 2026
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Keywords: n Artocarpus altilisn , Purification and characterisation, Natural fruit extracts, Browning control, Enzyme inhibition
Abstract
Breadfruit (Artocarpus altilis ) is a nutritious yet underutilised tropical fruit, and its postharvest value is limited by polyphenol oxidase (PPO)-mediated enzymatic browning. This study characterised breadfruit PPO and assessed natural fruit extracts as safe, sustainable anti-browning agents. PPO was purified by ammonium sulphate fractionation and chromatography, and its physicochemical properties and inhibition by natural fruit extracts were evaluated. Purified PPO was a 26.7 kDa monomer, optimally active at pH 5.0 and 40 °C, and specific for catechol, with Km and Vmax of 6.65 mM and 0.085 mg/min/mL. Residual activity fell below 10% at acidic pH (4.0 to 6.0) and at high temperatures (80 to 90 °C), indicating enzyme instability, while Mg2+, Ca2+ (5 mM), EDTA and glycine strongly inhibited it. Lemon extract inhibited PPO competitively, whereas onion extract acted via a non-competitive mechanism, suggesting active-site and allosteric inhibition. These findings provide a biochemical basis for understanding browning in breadfruit and support the use of natural, food-grade inhibitors for species-specific control of enzymatic browning during postharvest handling and processing.
References
Adeseko CJ, Sanni DM, Salawu SO, Kade IJ, Bamidele SO, Lawal OT (2021) Purification and biochemical characterization of polyphenol oxidase of African bush mango (Irvingia gabonensis) fruit peel. Biocatal Agric Biotechnol 36:102119. https://doi.org/10.1016/j.bcab.2021.102119
Adeseko CJ, Sanni DM, Lawal OT (2022) Biochemical studies of enzyme-induced browning of African bush mango (Irvingia gabonensis) fruit pulp. Prep Biochem Biotechnol 52(7):835–844. https://doi.org/10.1080/10826068.2021.1998113
Azzouzi N, Bouchaib A, Britel MR, Maurady A (2022) Characterization of polyphenol oxidase from blackberry thorny wild Rubus fruticosus and its inhibition using natural extracts. Curr Res Nutr Food Sci 10(3):1205–1221
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. https://doi.org/10.1016/0003-2697(76)90527-3
Bravo K, Osorio E (2016) Characterization of polyphenol oxidase from Cape gooseberry (Physalis peruviana L.) fruit. Food Chem 197:185–190. https://doi.org/10.1016/j.foodchem.2015.10.126
Demir D, Kabak S, Çağlayan K (2023) Purification and characterization of polyphenol oxidase in the fruits of Opuntia ficus-indica. Biology 12:1339
Geetha RY, Arulvel R, Selvarajan E (2014) Studies on kinetic parameters and biochemical characteristics of polyphenol oxidase purified from jackfruit (Artocarpus heterophyllus) waste. Int J Pharm Pharm Sci 6(9):473–478
Ismail HA, Richard I, Ramaiya SD, Zakaria MH, Lee SY (2023) Browning in relation to enzymatic activities and phytochemical content in terap peel (Artocarpus odoratissimus) during postharvest ripening. Horticulturae 9(1):57. https://doi.org/10.3390/horticulturae9010057
Janghu S, Kumar V, Yadav AK (2024) Post-harvest management of fruits and vegetables. Curr Perspect Agric Food Sci 7:125–148. https://doi.org/10.9734/bpi/cpafs/v7/7984E
Kim CY, Kim MJ, Lee MJ, Park I (2007) Inhibition of polyphenol oxidase and peach juice browning by onion extract. Food Sci Biotechnol 16:421–425
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. https://doi.org/10.1038/227680a0
Lawal OT, Fasan D, Ogundele OP, Olopoda AI, Omotoyinbo OV, Sanni DM (2025) Biochemical characterization of browning-induced enzyme in black-eyed Bambara groundnut (Vigna subterranea): Inhibition by natural fruit extracts. South Afr J Bot 180:134–143
Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666. https://doi.org/10.1021/ja01318a036
Moon KM, Kwon EB, Lee B, Kim CY (2020) Recent trends in controlling enzymatic browning of fruit and vegetable products. Molecules 25(12):2754. https://doi.org/10.3390/molecules25122754
Palma-Orozco G, Ortiz-Moreno A, Dorantes-Álvarez L, Sampedro JG, Nájera H (2011) Purification and partial biochemical characterization of polyphenol oxidase from mamey (Pouteria sapota). Phytochemistry 72(1):82–88. https://doi.org/10.1016/j.phytochem.2010.10.011
Peng X, Du C, Yu H, Zhao X, Zhang X, Wang X (2019) Purification and characterization of polyphenol oxidase from water yam. CyTA - J Food 17:676–684. https://doi.org/10.1080/19476337.2019.1634645
Shabnam N, Kahraman S (2023) Characterization of polyphenol oxidase from Eruca sativa. Sakarya Univ J Sci 27(4):887–894. https://doi.org/10.16984/saufenbilder.1174501
Sikora M, Świeca M, Franczyk M, Jakubczyk A, Bochnak J, Złotek U (2019) Biochemical properties of polyphenol oxidases from ready-to-eat lentil (Lens culinaris Medik.) sprouts and factors affecting their activities: A search for potent tools limiting enzymatic browning. Foods 8(5):154. https://doi.org/10.3390/foods8050154
Tao YM, Yao LY, Qin QY, Shen W (2013) Purification and characterization of polyphenol oxidase from jackfruit (Artocarpus heterophyllus) bulbs. J Agric Food Chem 61(51):12662–12669. https://doi.org/10.1021/jf403828e
Torres A, Aguilar-Osorio G, Camacho M, Basurto F, Navarro-Ocana A (2021) Characterization of polyphenol oxidase from purple sweet potato (Ipomoea batatas L. Lam) and its affinity towards acylated anthocyanins and caffeoylquinic acid derivatives. Food Chem 356:129709. https://doi.org/10.1016/j.foodchem.2021.129709
Author Information
Biochemistry Department, School of Life Sciences, Federal University of Technology, Akure , Nigeria