title: Optimization of Penicillium aurantiogriseum protease immobilization on magnetic nanoparticles for antioxidant peptides' obtainment.
abstract: This work reports an optimization of protease from Penicillium aurantiogriseum immobilization on polyaniline-coated magnetic nanoparticles for antioxidant peptides' obtainment derived from bovine casein. Immobilization process was optimized using a full two-level factorial design (24) followed by a response surface methodology. Using the derivative, casein was hydrolyzed uncovering its peptides that were sequenced and had antioxidant properties tested through (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) (ABTS) radical scavenging and hydrogen peroxide scavenging assays. Optimal conditions for immobilization were 2 hr of immobilization, offered protein amount of 200 µg/mL, immobilization pH of 6.3 and 7.3 hr of activation. Derivative keeps over 74% of its original activity after reused five times. Free and immobilized enzyme casein hydrolysates presented similar peptide mass fingerprints, and prevalent peptides could be sequenced. Hydrolysates presented more than 2.5× higher ROS scavenging activity than nonhydrolyzed casein, which validates the immobilized protease capacity to develop casein-derived natural ingredients with potential for functional foods.
authors.affiliation: a Keizo Asami immunopathology Laboratory (LIKA), Federal University of Pernambuco , Recife , PE , Brazil.
authors.forename: José Manoel Wanderley
authors.initials: JMW
authors.lastname: Duarte Neto
authors.affiliation: a Keizo Asami immunopathology Laboratory (LIKA), Federal University of Pernambuco , Recife , PE , Brazil.
authors.forename: Jackeline da Costa
authors.initials: JDC
authors.lastname: Maciel
authors.affiliation: b Northeastern strategic technology center (CETENE) , Recife , PE , Brazil.
authors.forename: Júlia Furtado
authors.initials: JF
authors.lastname: Campos
authors.affiliation: a Keizo Asami immunopathology Laboratory (LIKA), Federal University of Pernambuco , Recife , PE , Brazil.|c Biochemistry Department, Federal University of Pernambuco (UFPE) , Recife , PE , Brazil.
authors.forename: Luiz Bezerra de
authors.initials: LB
authors.lastname: Carvalho Junior
authors.affiliation: d University of Pernambuco, Serra Talhada Campus , Serra Talhada , PE , Brazil.
authors.forename: Daniela Araújo Viana
authors.initials: DAV
authors.lastname: Marques
authors.affiliation: e Faculty of Science, Education and Technology of Garanhuns, University of Pernambuco , Garanhuns , PE , Brazil.
authors.forename: Carolina de Albuquerque
authors.initials: CA
authors.lastname: Lima
authors.affiliation: a Keizo Asami immunopathology Laboratory (LIKA), Federal University of Pernambuco , Recife , PE , Brazil.|d University of Pernambuco, Serra Talhada Campus , Serra Talhada , PE , Brazil.
authors.forename: Ana Lúcia Figueiredo
authors.initials: ALF
authors.lastname: Porto
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002364:Caseins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D010407:Penicillium / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D002364:Caseins; D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D010455:Peptides; C475729:casein hydrolysate; D010447:Peptide Hydrolases
keywords: Bioactive peptides; functional foods; mass spectrometry analysis; protease immobilization; response surface methodology