title: Partial purification and immobilization/stabilization on highly activated glyoxyl-agarose supports of different proteases from flavourzyme.
abstract: The fractioning of some components and their immobilization of Flavourzyme, a commercial protease/aminopeptidase preparation, has been investigated to improve its specificity and stability. Adsorption of Flavourzyme on two ionic exchangers yielded two fractions with endoprotease activity and one fraction containing aminopeptidase activity. The use of an amine agarose gel has made it possible to purify a 43 kDa protein with only endoprotease activity. Immobilization of this endoprotease and the original Flavourzyme preparation onto glyoxyl-agarose provided derivatives that were more thermostable than their soluble counterparts. Tests using immobilized Flavourzyme and immobilized purified endoprotease for the hydrolysis of chickpea proteins showed that both preparations can be used for the production of protein hydrolysates and compare very favorably with the original crude Flavourzyme in terms of reducing the production of free amino acids. This was especially so in the case of immobilized endoprotease, which produced only 0.2% free amino acids. Keeping free amino acids content low is very important in protein hydrolysates for nutritional use to avoid excessive osmotic pressure.
authors.affiliation: Instituto de la Grasa, CSIC, Avenida Padre García Tejero 4, 41012 Sevilla, Spain. mdmar@cica.es
authors.forename: María del Mar
authors.initials: Mdel M
authors.lastname: Yust
authors.forename: Justo
authors.initials: J
authors.lastname: Pedroche
authors.forename: Manuel
authors.initials: M
authors.lastname: Alaiz
authors.forename: Julio
authors.initials: J
authors.lastname: Girón-Calle
authors.forename: Javier
authors.initials: J
authors.lastname: Vioque
authors.forename: César
authors.initials: C
authors.lastname: Mateo
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Guisan
authors.forename: Francisco
authors.initials: F
authors.lastname: Millan
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernandez-Lafuente
mesh_terms: D000327:Adsorption; D029863:Cicer / Q000737:chemistry; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D006038:Glyoxylates; D006868:Hydrolysis; D010940:Plant Proteins / Q000378:metabolism; D011492:Protein Hydrolysates; D012685:Sepharose
chemical_list: D004800:Enzymes, Immobilized; D006038:Glyoxylates; D010940:Plant Proteins; D011492:Protein Hydrolysates; C070399:glyoxyl agarose; D012685:Sepharose; D010450:Endopeptidases; C412470:flavourzyme