Production, one-step purification, and characterization of a keratinolytic protease from Serratia marcescens P3
2012
Bach, Evelise | Sant’Anna, Voltaire | Daroit, Daniel Joner | Corrêa, Ana Paula Folmer | Segalin, Jeferson | Brandelli, Adriano
Applications of keratinases demand efficient production and downstream processing. This work reports the production, purification and partial characterization of a keratinolytic protease from Serratia marcescens P3. The influence of temperature, initial pH and feather meal concentration on enzyme production was investigated using response surface methodology. Maximal protease production (12.5UmL⁻¹) was observed at 18–30°C, 10–24gL⁻¹ feather meal, and pH values of 6.5–8.5. The enzyme was recovered from culture supernatants using aqueous two-phase systems (ATPSs), and higher yields (68%) were obtained in polyethylene glycol (PEG) 4000-sodium citrate and PEG 4000-ammonium sulfate systems. These ATPS were effective for enzyme purification, resulting in an electrophoretically homogeneous protein of 53kDa. Sequencing of tryptic peptides identified this enzyme as a metalloprotease belonging to the serralysin family of metzincins. The purified enzyme showed optimal activity at 40–45°C and pH 6.5, being also active toward azokeratin.
Show more [+] Less [-]AGROVOC Keywords
Bibliographic information
This bibliographic record has been provided by National Agricultural Library