title: Characterisation of the aminopeptidase from non-germinated winter rape (Brassica napus L.) seeds.
abstract: Rapeseed plays a crucial role in food and fuel industry. Since aminopeptidases take part in many physiological processes in all organisms, it is important to learn their role and characteristics in economically relevant plants. Extracts of non-germinated winter rape seeds were screened for aminopeptidase activity. Substrate specificity, the influence of pH and temperature, as well as effect of protease inhibitors and chosen metal ions on the aminopeptidase activity were determined. The approximate molecular weight estimated by NATIVE-PAGE and SDS-PAGE electrophoresis was ∼60 kDa. The partially purified enzyme as well as the aminopeptidases present in crude extract cleaved preferentially Phe-pNA. The activity profiles toward several substrates were also determined. Maximum activity was observed at pH 6.5 and temperature of 40 °C for Phe-pNA as a substrate. Two visible picks in the pH profile toward Phe-pNA, together with other results (IEF) suggest the presence of more than one aminopeptidase, having similar molecular mass. Much lower activity and broad pH profiles were observed for Leu- and Ala-pNA as substrates.
authors.affiliation: Department of Organic Chemistry, Bioorganic Chemistry and Biotechnology, Faculty of Chemistry, Silesian University of Technology, 44-100 Gliwice, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Kania
authors.affiliation: Department of Organic Chemistry, Bioorganic Chemistry and Biotechnology, Faculty of Chemistry, Silesian University of Technology, 44-100 Gliwice, Poland. Electronic address: Danuta.Gillner@polsl.pl.
authors.forename: Danuta M
authors.initials: DM
authors.lastname: Gillner
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism*; D029688:Brassica napus / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D012639:Seeds / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000626:Aminopeptidases
keywords: Aminopeptidase; Characterisation; Inhibition; Non-germinated rape seeds; Substrate specificity