Optimization of Penicillium aurantiogriseum protease immobilization on magnetic nanoparticles for antioxidant peptides' obtainment

• Published in: Preparative biochemistry & biotechnology 2017-08, Vol.47 (7), p.644-654
• Main Authors: Duarte Neto, José Manoel Wanderley, Maciel, Jackeline da Costa, Campos, Júlia Furtado, Carvalho Junior, Luiz Bezerra de, Marques, Daniela Araújo Viana, Lima, Carolina de Albuquerque, Porto, Ana Lúcia Figueiredo
• Format: Article
• Language: English
• Subjects: Animals; Antioxidants; Antioxidants - chemistry; Antioxidants - metabolism; Antioxidants - pharmacology; Bioactive peptides; Casein; Caseins - chemistry; Caseins - metabolism; Caseins - pharmacology; Cattle; Design optimization; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Factorial design; Fingerprints; Food; functional foods; Functional foods & nutraceuticals; Fungi; Hydrogen peroxide; Hydrolysates; Hydrolysis; Immobilization; Magnetite Nanoparticles - chemistry; mass spectrometry analysis; Nanoparticles; Penicillium - enzymology; Peptide Hydrolases - chemistry; Peptide Hydrolases - metabolism; Peptides; Peptides - chemistry; Peptides - metabolism; Peptides - pharmacology; pH effects; Polyanilines; Protease; protease immobilization; Proteinase; Response surface methodology; Salts; Scavenging; Sulfonic acid
• ISSN: 1082-6068; 1532-2297
• DOI: 10.1080/10826068.2017.1292286

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 (2 4 ) 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.