Record Details

Influence of Storage Temperature on Total Peroxidase Activity in Crude Extracts from Picea abies L. Karst. Needles

Croatian International Relations Review

View Archive Info
 
 
Field Value
 
Title Influence of Storage Temperature on Total Peroxidase Activity in Crude Extracts from Picea abies L. Karst. Needles
Utjecaj temperature pohranjivanja na aktivnost ukupnih peroksidaza u nepročišćenom ekstraktu iglica smreke (Picea abies L. Karst.)
 
Creator Has-Schön, Elizabeta
Lepeduš, Hrvoje
Jerabek, Ljiljana
Cesar, Vera
 
Subject Picea abies; peroxidases; needles; activity; storage temperature
 
Description Peroxidases are enzymes with numerous functions in plant cells and are, therefore, interesting objects of current biochemical research. The purpose of this work was to investigate the effect of 30 days storage at different temperatures on the peroxidase activity in crude extracts from one-year old needles of Picea abies L. Karst. Total peroxidase activity was detected in the extract reaction with guaiacol and H2O2. Colour product was quantified by absorbance measurement at 470 nm. Measurements were performed daily in the first 10 days, and on the 12th, 14th, 16th, 18th and 30th day of the experiment. Extracts stored for 30 days were subjected to polyacrylamide gel electrophoresis, incubated with peroxidase substrates, and the resulting bands were quantified. Three different dependences on storage temperature were noticed. Extracts kept at –196 °C and –20 °C displayed no change in activity, samples kept at +24 °C and +37 °C decreased the activity, while those kept at +4 °C manifested a considerable rise in activity. Electropherogram analysis revealed two isoenzymes, with total activities resembling those obtained spectrophotometrically. There was no difference in isoenzyme appearance between samples. Recommendations regarding enzyme extract storage are given.
Peroksidaze su enzimi s brojnim funkcijama u biljnim stanicama. Stoga su ovi enzimi zanimljiv predmet suvremenih biokemijskih istraživanja. Svrha ovoga rada bila je ispitati utjecaj pohranjivanja kroz 30 dana pri različitim temperaturama na aktivnost ukupnih peroksidaza u nepročišćenom ekstraktu iz jednogodišnjih iglica smreke. Aktivnost ukupnih peroksidaza određivana je reakcijom proteinskoga ekstrakta s gvajakolom i H2O2. Količina obojenoga produkta proporcionalna je apsorbanciji na 470 nm. Mjerenja su vršena svaki dan prvih 10 dana, a nakon toga u slijedeće dane pokusa: 12., 14., 16., 18. i 30. Ekstrakti pohranjeni 30 dana bili su podvrgnuti nativnoj elektroforezi na poliakrilamidnome gelu te inkubirani sa supstratima peroksidaza, a intenzitet obojenja vrpci kvantitativno je određen. Uočene su tri različite ovisnosti o temperaturi pohranjivanja. Ekstrakti čuvani na –196 °C i –20 °C nisu promijenili aktivnost, uzorcima čuvanim na +24 °C i +37 °C aktivnost se smanjila, dok se onima čuvanim na +4 °C povećala. Analiza elektroferograma pokazala je postojanje dva izoenzima, pri čemu je zbroj njihovih pojedinačnih aktivnosti za svaku temperaturu pohranjivanja sličan onoj izmjerenoj spektrofotometrijski. Nije bilo razlike u broju i udjelu izoenzima između uzoraka. Preporuča se određ eni način pohranjivanja enzimskoga ekstrakta iz iglica smreke. .
 
Publisher Croatian Chemical Society
 
Date 2005-09-15
 
Type text
info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
 
Format application/pdf
 
Identifier http://hrcak.srce.hr/41
http://hrcak.srce.hr/file/41
 
Source Croatica Chemica Acta; Vol.78 No.3; ISSN 0011-1643 (Print); ISSN 1334-417X (Online)
 
Language en
 
Rights info:eu-repo/semantics/openAccess
Parts of the contents of Croat. Chem. Acta (e. g. figures or tables) may be reproduced without prior permission, provided reference is made to their source.