Artículo
Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
Minchiotti, Mariana Cecilia; Scalambro, Maria Belen
; Vargas, Laura Inés; Coronel, Carlos Enrique
; Madoery, Ricardo Roman
Fecha de publicación:
04/04/2008
Editorial:
Elsevier Science Inc
Revista:
Enzyme and Microbial Technology
ISSN:
0141-0229
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
A protein with phospholipase A2 (PLA2) activity was isolated from soybean (Glycine max) seeds. The affinity column chromatography techniques by using Cibacron Blue and immobilized substrate onto Eupergit C, were important steps in the purification protocol. The electrophoretic mobility showed by the purified enzyme, agreed with a molecular mass of 14 kDa. The PLA2 activity against liposomes was determined by measurement of apparent absorbance changes at 340 nm. Also, high performance thin layer chromatography (HPTLC) analysis confirmed selective hydrolysis at the sn-2 position of soybean phospholipids. PLA2 exhibited millimolar calcium dependence and had slightly alkaline pH optimum. The enzyme was stimulated by auxins and completely inactivated by ammonium sulfate. Furthermore, it was irreversibly inactivated by p-bromo-phenacyl bromide, the specific inhibitor of secretory PLA2s. In addition, Glycine max PLA2 showed high stability against heat treatment and organic solvents. The enzyme showed activity toward multilamellar vesicles of soybean phospholipids, with a Vmax = 950 U/mg and a KM = 0.78 mM. The hyperbolic behavior observed was coherent with a hopping mode of action, one of the two characteristic interfacial mechanisms of PLA2s. All these data agree with the expected properties for a secretory PLA2 being the first soybean enzyme of this type. The new PLA2 developed lipolytic activity on a water in oil microemulsion reaction system, which is suitable for lysophospholipid production. These lysoderivatives are valuable biosurfactants for food and pharmaceutical industries.
Palabras clave:
Phospholipase A2
,
Properties
,
Purification
,
Soybean
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Articulos(IIBYT)
Articulos de INSTITUTO DE INVESTIGACIONES BIOLOGICAS Y TECNOLOGICAS
Articulos de INSTITUTO DE INVESTIGACIONES BIOLOGICAS Y TECNOLOGICAS
Citación
Minchiotti, Mariana Cecilia; Scalambro, Maria Belen; Vargas, Laura Inés; Coronel, Carlos Enrique; Madoery, Ricardo Roman; Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties; Elsevier Science Inc; Enzyme and Microbial Technology; 42; 5; 4-4-2008; 389-394
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