A mixture of at least three proteolytic activities including an extracellular serine protease. In general, serine proteases display a wide range of substrate specificities, which are believed to be mediated by an active site composed of one Asp, one His, and a Ser residue in the molecule. This enzyme prefers to hydrolyze peptide bonds on the carboxyl side of glutamic or aspartic acid.
Especificidad
Mezcla de como mínimo tres actividades proteolíticas, entre ellas una serín proteasa extracelular. En general, las proteasas de serina exhiben una amplia variedad de especificidades de sustrato, que se creen mediadas por un centro activo compuesto de un resto de Asp, uno de His y uno de Ser en la molécula. Esta enzima prefiere hidrolizar los enlaces peptídicos del lado carboxilo de los ácidos glutámico o aspártico.
Aplicación
Esta enzima es más activa a un intervalo de pH más elevado que la proteasa alcalina conocida, mostrando actividad proteolítica incluso en disolución de NaOH 0,2 N. Esta enzima es útil para la proteólisis de proteínas insolubles y para la investigación de la estructura de las proteínas.
La proteasa se utiliza normalmente en los procedimientos de aislamiento de ácidos nucleicos en incubaciones de 0,5 a 3,0 horas complementadas con dodecilsulfatosódico al 0,2 % y EDTA 10mM.
Suitable for:
Nucleic acid isolation procedures in incubations
Degrade protein during nucleic acid purification
Proteolysis of insoluble protein
Structural protein studies
Propiedades físicas
Completamente inactivada al calentarse por encima de 80 °C durante 15 a 20 minutos.
Definición de unidad
One unit will hydrolyze casein to produce color equivalent to 1.0 μmole (181 μg) of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).
Nota de preparación
Recolectada del caldo de cultivo de S. griseus.
Nota de análisis
The protease is incubated for 10 minutes at pH 7.5 at 37°C in a 6 ml reaction volume containing 0.54% casein and 0.041 M potassium phosphate buffer. The reaction is stopped by the addition of 5.0 ml of 0.11 M trichloroacetic acid.
Otras notas
This protease is completely inactivated by heating above 80°C for 15-20 minutes. This enzyme is more active at a higher pH range, showing the proteolytic activity even in 0.2N NaOH solution.
Bioscience, biotechnology, and biochemistry, 58(3), 470-474 (1994-03-01)
SAP, an extracellular alkaline serine protease produced by Streptomyces sp. YSA-130, was purified to homogeneity by CM-Sephadex column chromatography and crystallization. The enzyme was a monomeric protein with a molecular weight of 19,000 as estimated by SDS-PAGE and gel filtration.
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