INVESTIGADORES
MUÑOZ Fernando Felipe
congresos y reuniones científicas
Título:
PURIFICATION OF AN ASPARTIC PROTEINASE FROM SOLANUM ELAEAGNIFOLIUM FRUITS (SEAP-1)
Autor/es:
TORRES NÁGERA M.; MUÑOZ F.; LÓPEZ LÓPEZ L.; DE LA CRUZ GALICIA MG; SILVIA BELMARES SY; GUEVARA M.
Lugar:
Rosario
Reunión:
Congreso; L Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2014
Resumen:
Solanum
elaeagnifolium is an endemic plant
from the northeast of Mexico. This plant in some places of Mexico has been used
for decades in the manufacture of artisanal filata-type asadero cheese. The
milk-clotting activity of S. elaeagnifolium has been attributed to
aspartic proteinases. The aim of this work was to purify aspartic proteinases
from fruits of S. elaeagnifolium. Purification was performed from
lyophilized fruits of S. eleagnifolium by ammonium sulfate
precipitation; ion exchange chromatography and Pepstatin A affinity
chromatography. SDS- PAGE analysis of proteins eluted from the affinity column
shown only one protein band corresponding to 59 kDa., approximately. Identity
of this protein band as aspartic proteinase was demonstrated by Western blot
analysis, using as primary antibody IgG-anti StAP1 (Solanum tuberosum
aspartic proteinase 1). Results obtained from gel filtration chromatography
(Superose 12) demonstrates that, isolated SeAP1 is a monomeric enzyme,
with an estimated molecular weight of 52 kDa., approximately. Additionally, we
determined that, like almost monomeric plant APs, SeAP-1 exerts
cytotoxic activity towards plant pathogens in a dose- dependent manner. These
results suggest the presence of saposin- like domain into the sequence of the
mature SeAP-1 and therefore, new biotechnological applications for SeAP-1.