Purification of hydroperoxide lyase from green bell pepper (Capsicum annuum L.) fruits for the generation of C6-aldehydes in vitro.
The aim of this work was to compare the efficiency of different extracts of hydroperoxide lyase from green bell peppers in producing aldehydes: a crude extract, a chloroplastic fraction, and a purified enzyme were investigated. From a crude extract, the HPO lyase was purified by ion-exchange chromatography with a 22.3-fold increase in purification factor. Analysis by SDS-PAGE electrophoresis under denaturating conditions showed only one protein with a molecular weight of 55 kDa, whereas size-exclusion chromatography indicated a molecular weight of 170 kDa. A maximum of 7500 mg of aldehydes per g of protein was obtained with the purified enzyme within 20 min of bioconversion compared to 392 and 88 mg of aldehydes per g of protein within 50 and 60 min, respectively, for the chloroplast fraction and the crude extract.
Husson
F
F
Laboratoire de Biotechnologie, Equipe Ingénierie Moléculaire et Sensorielle des Aliments, ENSBANA, Université de Bourgogne, Dijon, France. Florence.Husson@u-bourgogne.fr
Belin
J M
JM
D000446:Aldehyde-Lyases / Q000302:isolation & purification*; D000447:Aldehydes / Q000378:metabolism*; D002212:Capsicum / Q000201:enzymology*; D002736:Chloroplasts / Q000201:enzymology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D008041:Linoleic Acids / Q000378:metabolism; D008042:Linolenic Acids / Q000378:metabolism; D008054:Lipid Peroxides / Q000378:metabolism; D008970:Molecular Weight; D010936:Plant Extracts / Q000737:chemistry; D011489:Protein Denaturation
D000447:Aldehydes; D008041:Linoleic Acids; D008042:Linolenic Acids; D008054:Lipid Peroxides; D010936:Plant Extracts; C038649:13-hydroperoxy-9,11-octadecadienoic acid; C057375:13-hydroperoxy-9,11,15-octadecatrienoic acid; D003577:Cytochrome P-450 Enzyme System; D000446:Aldehyde-Lyases; C054540:hydroperoxide lyase