title: A novel malate dehydrogenase from Ceratonia siliqua L. seeds with potential biotechnological applications.
abstract: A novel malate dehydrogenase (MDH; EC 3.1.1.1.37), hereafter MDHCs, from Ceratonia siliqua seeds, commonly known as Carob tree, was purified by using ammonium sulphate precipitation, ion exchange chromatography on SteamLine SP and gel-filtration. The molecular mass of the native protein, obtained by analytical gel-filtration, was about 65 kDa, whereas, by using SDS-PAGE analysis, with and without reducing agent, was 34 kDa. The specific activity of purified MDHCs (0.25 mg/100 g seeds) was estimated to be 188 U/mg. The optimum activity of the enzyme is at pH 8.5, showing a decrease in the presence of Ca(2+), Mg(2+) and NaCl. The N-terminal sequence of the first 20 amino acids of MDHCs revealed 95 % identity with malate dehydrogenase from Medicago sativa L. Finally, the enzymatic activity of MDHCs was preserved even after absorption onto a PVDF membrane. To our knowledge, this is the first contribution to the characterization of an enzyme from Carob tree sources.
authors.affiliation: Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Caserta, Italy.
authors.forename: Clelia
authors.initials: C
authors.lastname: Muccio
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Guida
authors.forename: Amalia
authors.initials: A
authors.lastname: Di Petrillo
authors.forename: Valeria
authors.initials: V
authors.lastname: Severino
authors.forename: Antimo
authors.initials: A
authors.lastname: Di Maro
mesh_terms: D000042:Absorption; D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D007887:Fabaceae / Q000737:chemistry / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008291:Malate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D012639:Seeds / Q000737:chemistry / Q000201:enzymology; D016415:Sequence Alignment
chemical_list: D008291:Malate Dehydrogenase