Abstract
Background: Polyphenol Oxidase (PPO) belongs to the oxidoreductase enzyme family.
Methods: Here, PPO was purified from potato using Sepharose 4B-L-tyrosine-p-aminobenzoic acid affinity chromatography. It determined the interactions between some phenolic acids and the enzyme.
Results: The enzyme was obtained with a specific activity of 15333.33 EU/mg protein and 7.87- fold purification. It was found that phenolic acids exhibited inhibitory properties for PPO. The IC50 values of the phenolic acids were found in the range of 0.36-2.12 mM, and their Ki values were found in the range of 0.28± 0.07-1.72±0.32 mM. It was determined that all studied compounds displayed a competitive inhibition effect. Among these compounds, 3-hydroxybenzoic acid was found to be the most effective PPO inhibitor (Ki: 0.28±0.07 mM).
Conclusion: Investigating the inhibition kinetics of the enzyme will simplify the testing of PPO inhibitor candidates.
Keywords: Affinity chromatography, inhibition, phenolic acids, purification, polyphenol oxidase, hydroxybenzoic acid.
Graphical Abstract
[http://dx.doi.org/10.1080/10942912.2015.1012725]
[http://dx.doi.org/10.1016/j.phytochem.2010.10.011] [PMID: 21087780]
[http://dx.doi.org/10.1016/j.foodchem.2009.04.013]
[http://dx.doi.org/10.1016/j.foodchem.2003.12.025]
[http://dx.doi.org/10.1016/j.foodchem.2009.03.088]
[http://dx.doi.org/10.1111/j.1365-2621.2001.tb04594.x]
[http://dx.doi.org/10.1080/14756360310001640472] [PMID: 15449733]
[http://dx.doi.org/10.1002/jbt.21935] [PMID: 28557170]
[http://dx.doi.org/10.1016/j.ijbiomac.2018.08.004] [PMID: 30077669]
[http://dx.doi.org/10.1002/ardp.201800263] [PMID: 30478943]
[http://dx.doi.org/10.2174/0929866526666190301115122] [PMID: 30827223]
[http://dx.doi.org/10.1002/bab.1781] [PMID: 31135076]
[http://dx.doi.org/10.1016/j.jnutbio.2008.07.009] [PMID: 18926684]
[http://dx.doi.org/10.1016/j.biortech.2016.01.116] [PMID: 26868149]
[http://dx.doi.org/10.1021/jp3116319] [PMID: 23418927]
[http://dx.doi.org/10.1016/j.bionut.2014.07.010]
[http://dx.doi.org/10.3390/molecules15117985] [PMID: 21060304]
[http://dx.doi.org/10.1590/S1984-82502013000300002]
[http://dx.doi.org/10.1093/nar/gkr1251] [PMID: 22187155]
[http://dx.doi.org/10.1016/j.bbrc.2018.08.043] [PMID: 30217455]
[http://dx.doi.org/10.1016/j.foodchem.2004.04.005]
[http://dx.doi.org/10.1016/0003-2697(76)90527-3] [PMID: 942051]
[http://dx.doi.org/10.1104/pp.81.2.614] [PMID: 16664865]
[http://dx.doi.org/10.1038/227680a0] [PMID: 5432063]
[http://dx.doi.org/10.1007/s12010-019-03009-x] [PMID: 30980289]
[http://dx.doi.org/10.1111/jphp.13144] [PMID: 31347707]
[http://dx.doi.org/10.1016/j.pharep.2019.02.012] [PMID: 31109643]
[http://dx.doi.org/10.1016/j.etap.2019.103195] [PMID: 31125830]
[http://dx.doi.org/10.1021/ja01318a036]
[http://dx.doi.org/10.1073/pnas.1523575113] [PMID: 26976571]
[http://dx.doi.org/10.1016/j.phytochem.2006.08.006] [PMID: 16973188]
[http://dx.doi.org/10.1002/jsfa.2144]
[http://dx.doi.org/10.1016/S0969-806X(97)00318-6]
[http://dx.doi.org/10.1111/jfbc.12770] [PMID: 31353556]
[http://dx.doi.org/10.1021/acs.jafc.7b03210] [PMID: 28812349]
[http://dx.doi.org/10.1016/j.foodchem.2009.07.013]
[http://dx.doi.org/10.1016/j.foodchem.2016.09.061] [PMID: 27719900]
[http://dx.doi.org/10.3390/molecules24101922] [PMID: 31109085]
[http://dx.doi.org/10.1080/1475636032000141890] [PMID: 16119202]
[http://dx.doi.org/10.1016/j.bioorg.2015.10.003] [PMID: 26496408]