Purification of polyphenol oxidase from borage (Trachystemon orientalis L.) by using three-phase partitioning and investigation of kinetic properties.
In this study a Polyphenol oxidase from borage plant was purified with 3.59-fold enrichment in the specific activity and 68.75% recovery of the total activity by using three-phase partitioning purification technique for the first time. Its molecular weight was found around 80kDa with sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature values of the enzyme for the used four substrates ranged between the pH 5.0-7.5 and 5-30°C. The kcat/Kmvalues showed that the enzyme has the greatest reactivity toward caffeic acid among the substrates used. Ascorbic acid, l-cysteine and sodium metabisulfite markedly inhibited borage polyphenol oxidase activity.
Department of Chemistry, Faculty of Science and Arts, Sakarya University, Serdivan-Sakarya, 54187, Turkey. Electronic address: ealici@sakarya.edu.tr.
Esma Hande
EH
Alici
Department of Chemistry, Faculty of Science and Arts, Sakarya University, Serdivan-Sakarya, 54187, Turkey. Electronic address: garabaci@sakarya.edu.tr.
Gulnur
G
Arabaci
D028082:Boraginaceae / Q000201:enzymology*; D004156:Catechol Oxidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D011232:Chemical Precipitation; D004791:Enzyme Inhibitors / Q000737:chemistry; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D059625:Liquid-Liquid Extraction; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D011489:Protein Denaturation; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D010940:Plant Proteins; D004156:Catechol Oxidase
Polyphenol oxidase; Three-phase partitioning; Trachystemon orientalis L.