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Abstract:

Solid state fermentation cultures of Aspergillus oryzae NRRL 3485 on moistened wheat bran produced high levels of α-amylase as judged by the specific activity of water extracts (500-700 units/mg protein) and by the enzyme concentration (around 0.15 g/L ) in those extracts. The purification of α-amylase by affinity chromatography on Concanavalin A-Sepharose is described. A first DEAE-Sepharose chromatography step (binding of the enzyme contained in the water extract and further elution with 0.2 M NaCl, pH 5) was necessary in order to remove contaminants that hinder the binding of the glycoprotein α-amylase to the lectin. Elution was performed with 7.5 mM α-D-methylmannoside. The enzyme was obtained highly concentrated and purified with a specific activity of 2000 units/mg protein and a recovery of around 60%, free of α-glucosidase, amyloglucosidase and color contaminants. The purity of the enzyme preparation was assessed through nondenaturing gel electrophoresis and sucrose gradient centrifugation. An improvement of the performance of the purification procedure is presented in which the maximum capacities of both the anion exchanger and the lectin matrices were exploited.

Registro:

Documento: Artículo
Título:Effective purification procedure of Aspergillus oryzae α-amylase from solid state fermentation cultures including Concanavalin A-Sepharose
Autor:Terebiznik, M.R.; Pilosof, A.M.R.; Moreno, S.
Filiación:Departamento de Industrias, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Depto. de Quim. Biológica, Fac. de Ciencias Exactas y Naturales, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Departmento de Ing. Química, Facultad de Ingeniería, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Departamento de Industrias, Fac. de Ciencias Exactas y Naturales, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Idioma: Inglés
Palabras clave:Aspergillus oryzae; Triticum aestivum
Año:1996
Volumen:19
Número:5
Página de inicio:341
Página de fin:354
Título revista:Journal of Food Biochemistry
Título revista abreviado:J. Food Biochem.
ISSN:01458884
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01458884_v19_n5_p341_Terebiznik

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Citas:

---------- APA ----------
Terebiznik, M.R., Pilosof, A.M.R. & Moreno, S. (1996) . Effective purification procedure of Aspergillus oryzae α-amylase from solid state fermentation cultures including Concanavalin A-Sepharose. Journal of Food Biochemistry, 19(5), 341-354.
Recuperado de https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01458884_v19_n5_p341_Terebiznik [ ]
---------- CHICAGO ----------
Terebiznik, M.R., Pilosof, A.M.R., Moreno, S. "Effective purification procedure of Aspergillus oryzae α-amylase from solid state fermentation cultures including Concanavalin A-Sepharose" . Journal of Food Biochemistry 19, no. 5 (1996) : 341-354.
Recuperado de https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01458884_v19_n5_p341_Terebiznik [ ]
---------- MLA ----------
Terebiznik, M.R., Pilosof, A.M.R., Moreno, S. "Effective purification procedure of Aspergillus oryzae α-amylase from solid state fermentation cultures including Concanavalin A-Sepharose" . Journal of Food Biochemistry, vol. 19, no. 5, 1996, pp. 341-354.
Recuperado de https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01458884_v19_n5_p341_Terebiznik [ ]
---------- VANCOUVER ----------
Terebiznik, M.R., Pilosof, A.M.R., Moreno, S. Effective purification procedure of Aspergillus oryzae α-amylase from solid state fermentation cultures including Concanavalin A-Sepharose. J. Food Biochem. 1996;19(5):341-354.
Available from: https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01458884_v19_n5_p341_Terebiznik [ ]