Production, one-step purification, and characterization of a keratinolytic protease from Serratia marcescens P3

• DOI: 10.1016/j.procbio.2012.10.007
• Publication Date: 2012-10-27T03:31:44Z
• Authors: Evelise Bach; Voltaire Sant’Anna; Daniel Joner Daroit; Ana Paula Folmer ...; Jeferson Segalin; Adriano Brandelli
• Keywords: Keratinase; Ammonium sulfate; Feather meal; Sodium dodecyl sulfate

Applications of keratinases demand efficient production and downstream processing. This work reports the production, purification partial characterization a keratinolytic protease from Serratia marcescens P3. The influence temperature, initial pH feather meal concentration on enzyme was investigated using response surface methodology. Maximal (12.5 U mL−1) observed at 18–30 °C, 10–24 g L−1 meal, values 6.5–8.5. recovered culture supernatants aqueous two-phase systems (ATPSs), higher yields (68%) were obtained in polyethylene glycol (PEG) 4000-sodium citrate PEG 4000-ammonium sulfate systems. These ATPS effective for purification, resulting an electrophoretically homogeneous protein 53 kDa. Sequencing tryptic peptides identified this as metalloprotease belonging to serralysin family metzincins. purified showed optimal activity 40–45 °C 6.5, being also active toward azokeratin.