Horseradish esterases: detection, purification and identification (CROSBI ID 237527)
Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija
Podaci o odgovornosti
Leščić Ašler, Ivana ; Peharec Štefanić, Petra ; Balen, Biljana ; Allmaier, Günter ; Marchetti- Deschmann, Martina ; Kojić-Prodić, Biserka
engleski
Horseradish esterases: detection, purification and identification
Our goal is to characterize esterases from horseradish tissues and assign their physiological roles. In the present study we focused on isolation, purification and identification of esterases from different horseradish tissues: plantlets and two tumor tissue lines. Horizontal IEF system enabled separation of six esterase isoforms with quite different pI values as well as with pronounced differences in expression levels among analyzed tissues. Esterases were extracted, fractionated by means of cation exchange chromatography, and analyzed by planar gel electrophoresis (SDS– PAGE) and isoelectrical focusing (IEF), UV/Vis spectroscopy, MALDI mass spectrometry (MS) and MALDI-MS/MS. Several chromatographic strategies were applied for esterase purification and characterization. Two subsequent cation exchange chromatographic steps based on SP-Sepharose FF material, followed by in-solution digestion combined with MALDI-MS and MS/MS proved to be the best strategy for identification of two esterase proteins, namely Pectinesterase/pectinesterase inhibitor 18 and GDSL esterase/lipase ESM1.
Armoracia lapathifolia ; Protein identification ; Protein purification ; Electrophoresis ; Mass spectrometry ; Esterase
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Podaci o izdanju
130 (1)
2017.
13-24
objavljeno
0167-6857
10.1007/s11240-017-1200-0
Povezanost rada
Kemija, Biologija, Biotehnologija