Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos

Abstract One important functional food ingredient today, valued for its health properties and ability to prevent disease, is bee pollen, which comprises a combination of nectar, pollen from plants, and the secretions of bees. In this research, the tyrosinase (TYR) inhibiting abilities of the peptide...

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Main Authors: Papassara Sangtanoo, Piroonporn Srimongkol, Tanatorn Saisavoey, Songchan Puthong, Anumart Buakeaw, Rutairat Suttisuwan, Marisa Jatupornpipat, Wittaya Pimtong, Onrapak Reamtong, Aphichart Karnchanatat
Format: Article
Language:English
Published: Nature Portfolio 2024-12-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-024-81495-8
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author Papassara Sangtanoo
Piroonporn Srimongkol
Tanatorn Saisavoey
Songchan Puthong
Anumart Buakeaw
Rutairat Suttisuwan
Marisa Jatupornpipat
Wittaya Pimtong
Onrapak Reamtong
Aphichart Karnchanatat
author_facet Papassara Sangtanoo
Piroonporn Srimongkol
Tanatorn Saisavoey
Songchan Puthong
Anumart Buakeaw
Rutairat Suttisuwan
Marisa Jatupornpipat
Wittaya Pimtong
Onrapak Reamtong
Aphichart Karnchanatat
author_sort Papassara Sangtanoo
collection DOAJ
description Abstract One important functional food ingredient today, valued for its health properties and ability to prevent disease, is bee pollen, which comprises a combination of nectar, pollen from plants, and the secretions of bees. In this research, the tyrosinase (TYR) inhibiting abilities of the peptides derived from bee pollen protein hydrolysates are investigated. Various proteases were utilized to generate these peptides, followed by testing at different concentrations. Tyrosinase inhibition activity was detected in all cases, while the hydrolysate drawn from 5.0% w/v neutrase exhibited the best IC50 value and was thus investigated further via ultrafiltration to separate the active fractions. The highest potential for tyrosinase inhibition was recorded for the fractions below 0.65 kDa. Subsequent purification steps via SEC and RP-HPLC led to the identification of the VDGYPAAGY (named VY-9) peptide via LC-Q-TOF-MS/MS in fraction F1–2, known for its non-toxic and hydrophobic characteristics albeit poor water solubility. The synthesized VY-9 peptide demonstrated competitive inhibition, with IC50 values of 0.55 ± 0.03 µM for mono-phenolase and 2.54 ± 0.06 µM for di-phenolase activities, as confirmed by molecular docking analysis revealing dominant hydrogen bond interactions with TYR. Effective concentrations of 0.2–1.6 µM of VY-9 showed negligible cytotoxicity in B16F10 cells. Melanin synthesis suppression was examined via qRT-PCR, and western blot in MITF, TYR, TRP-1, and TRP-2. Cell death in zebrafish embryos was evaluated in vivo using a toxicity assay which revealed no significant influence from VY-9, while anti-melanogenic effects were observed when the concentration was 4 µM, suggesting bee pollen-derived peptides’ potential in cosmetic and pharmaceutical depigmentation applications.
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spelling doaj-art-7c0e05906e0b4f7ea8577154c9481f3e2025-08-20T02:39:35ZengNature PortfolioScientific Reports2045-23222024-12-0114112210.1038/s41598-024-81495-8Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryosPapassara Sangtanoo0Piroonporn Srimongkol1Tanatorn Saisavoey2Songchan Puthong3Anumart Buakeaw4Rutairat Suttisuwan5Marisa Jatupornpipat6Wittaya Pimtong7Onrapak Reamtong8Aphichart Karnchanatat9Center of Excellence in Bioconversion and Bioseparation for Platform Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn UniversityCenter of Excellence in Bioconversion and Bioseparation for Platform Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn UniversityCenter of Excellence in Bioconversion and Bioseparation for Platform Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn UniversityCenter of Excellence in Bioconversion and Bioseparation for Platform Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn UniversityCenter of Excellence in Bioconversion and Bioseparation for Platform Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn UniversityBiodiversity and Sustainable Utilization Research Unit, Department of Biology, Faculty of Science and Technology, Rajamangala University of Technology KrungthepDepartment of Biology, Faculty of Science, King Mongkut’s Institute of TechnologyNano Environmental and Health Safety Research Team, National Science and Technology Development Agency (NSTDA)Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol UniversityCenter of Excellence in Bioconversion and Bioseparation for Platform Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn UniversityAbstract One important functional food ingredient today, valued for its health properties and ability to prevent disease, is bee pollen, which comprises a combination of nectar, pollen from plants, and the secretions of bees. In this research, the tyrosinase (TYR) inhibiting abilities of the peptides derived from bee pollen protein hydrolysates are investigated. Various proteases were utilized to generate these peptides, followed by testing at different concentrations. Tyrosinase inhibition activity was detected in all cases, while the hydrolysate drawn from 5.0% w/v neutrase exhibited the best IC50 value and was thus investigated further via ultrafiltration to separate the active fractions. The highest potential for tyrosinase inhibition was recorded for the fractions below 0.65 kDa. Subsequent purification steps via SEC and RP-HPLC led to the identification of the VDGYPAAGY (named VY-9) peptide via LC-Q-TOF-MS/MS in fraction F1–2, known for its non-toxic and hydrophobic characteristics albeit poor water solubility. The synthesized VY-9 peptide demonstrated competitive inhibition, with IC50 values of 0.55 ± 0.03 µM for mono-phenolase and 2.54 ± 0.06 µM for di-phenolase activities, as confirmed by molecular docking analysis revealing dominant hydrogen bond interactions with TYR. Effective concentrations of 0.2–1.6 µM of VY-9 showed negligible cytotoxicity in B16F10 cells. Melanin synthesis suppression was examined via qRT-PCR, and western blot in MITF, TYR, TRP-1, and TRP-2. Cell death in zebrafish embryos was evaluated in vivo using a toxicity assay which revealed no significant influence from VY-9, while anti-melanogenic effects were observed when the concentration was 4 µM, suggesting bee pollen-derived peptides’ potential in cosmetic and pharmaceutical depigmentation applications.https://doi.org/10.1038/s41598-024-81495-8Bee pollenProtein hydrolysateTyrosinase inhibitory peptidesMelanogenesisB16F10 mouse melanoma cellsZebrafish
spellingShingle Papassara Sangtanoo
Piroonporn Srimongkol
Tanatorn Saisavoey
Songchan Puthong
Anumart Buakeaw
Rutairat Suttisuwan
Marisa Jatupornpipat
Wittaya Pimtong
Onrapak Reamtong
Aphichart Karnchanatat
Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
Scientific Reports
Bee pollen
Protein hydrolysate
Tyrosinase inhibitory peptides
Melanogenesis
B16F10 mouse melanoma cells
Zebrafish
title Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
title_full Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
title_fullStr Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
title_full_unstemmed Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
title_short Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
title_sort bee pollen peptides as potent tyrosinase inhibitors with anti melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
topic Bee pollen
Protein hydrolysate
Tyrosinase inhibitory peptides
Melanogenesis
B16F10 mouse melanoma cells
Zebrafish
url https://doi.org/10.1038/s41598-024-81495-8
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