dc.contributor.author | Onez, Zehra | |
dc.contributor.author | Karakus, Emine | |
dc.contributor.author | Pekyardimci, Sule | |
dc.date.accessioned | 2019-07-27T12:10:23Z | |
dc.date.accessioned | 2019-07-28T10:15:16Z | |
dc.date.available | 2019-07-27T12:10:23Z | |
dc.date.available | 2019-07-28T10:15:16Z | |
dc.date.issued | 2008 | |
dc.identifier.issn | 0145-8884 | |
dc.identifier.issn | 1745-4514 | |
dc.identifier.uri | https://dx.doi.org/10.1111/j.1745-4514.2008.00178.x | |
dc.identifier.uri | https://hdl.handle.net/20.500.12418/10373 | |
dc.description | WOS: 000256156800008 | en_US |
dc.description.abstract | Polyphenol oxidase (PPO) was extracted from the Izmir grape fruit (Vitis vinifera L.) and its characteristics were studied. The PPO enzyme was purified 2.2-fold by (NH4)(2)SO4 precipitation and 26.1-fold by gel filtration chromatography. This partially purified enzyme obtained from dialysis migrated as one band with catechol substrate, four bands with Coomassie brilliant R-250 (NATIVE-PAGE) native system and as six bands of 34, 30, 22, 18, 16 and 15 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The sample obtained from dialysis after ammonium sulfate precipitation was used for the characterization of partially purified enzyme. Substrate specificity experiments were carried out with catechol, 4-methyl catechol, L-tyrosine, pyrogallol, p-cresol, gallic acid and caffeic acid. Michael-Menten constant and maximum velocity values were 3.65 mM and 1,971.6 Delta A/dak, respectively, with catechol substrate. Optimum pH for the PPO enzyme were determined at 7.2, 7.0, 7.4, 7.2, 6,4, 6.7 and 5.7 using catechol, 4-methyl catechol, L-tyrosine, pyrogallol, p-cresol, gallic acid and caffeic acid substrates, respectively. Optimum temperature for maximum PPO activity was 25C for catechol. Heat inactivation studies showed a significiant decrease in enzymatic activity at temperatures above 45C. Activation energy was calculated to be 12.4 kcal/mol. The most potent inhibitors for the grape PPO were thiourea, sodium diethyldithiocarbamate and sodium azide. Inhibition constants were calculated for L-ascorbic acid, L-cysteine and beta-mercaptoethanol at the optimum pH of PPO activity. | en_US |
dc.language.iso | eng | en_US |
dc.publisher | WILEY | en_US |
dc.relation.isversionof | 10.1111/j.1745-4514.2008.00178.x | en_US |
dc.rights | info:eu-repo/semantics/closedAccess | en_US |
dc.title | Izmir grape polyphenol oxidase (Vitis vinifera L.): Partial purification and some kinetic properties | en_US |
dc.type | article | en_US |
dc.relation.journal | JOURNAL OF FOOD BIOCHEMISTRY | en_US |
dc.contributor.department | [Karakus, Emine] Yildiz Tech Univ, Dept Chem, Fac Sci & Arts, Istanbul, Turkey -- [Onez, Zehra] Cumhuriyet Univ, Dept Biochem, Fac Med, Sivas, Turkey -- [Pekyardimci, Sule] Ankara Univ, Dept Chem, Fac Sci, TR-06100 Ankara, Turkey | en_US |
dc.identifier.volume | 32 | en_US |
dc.identifier.issue | 3 | en_US |
dc.identifier.endpage | 414 | en_US |
dc.identifier.startpage | 396 | en_US |
dc.relation.publicationcategory | Makale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı | en_US |