title: Purification of serratiopeptidase from Serratia marcescens NRRL B 23112 using ultrasound assisted three phase partitioning.
abstract: The ultrasound assisted three phase partitioning (UATPP) is a novel bioseparation method for separation and purification of biomolecules. In the present work, UATPP was investigated for the first time for purification of serratiopeptidase from Serratia marcescens NRRL B 23112. Effect of various process parameters such as ammonium sulphate saturation, t-butanol to crude extract ratio, pH, ultrasonic frequency, ultrasonic intensity, duty cycle and irradiation time were evaluated and optimized. The optimized conditions were found to be as follows: ammonium sulphate saturation 30% (w/v), pH 7.0, t-butanol to crude ratio 1:1 (v/v), ultrasound frequency 25 kHz, ultrasound intensity 0.05 W/cm(2), duty cycle 20% and irradiation time 5 min. The maximum purity and recovery obtained from UATPP was 9.4-fold and 96% respectively as compared to the three phase partitioning (TPP) (4.2-fo ld and 83%). Also the process time for UATPP was significantly reduced to 5 min from 1h as compared to TPP. The results indicate that, UATPP is an efficient technique for the purification of serratiopeptidase with maximum purity, recovery and reduced processing time.
authors.affiliation: Department of Chemical Engineering, Institute of Chemical Technology, Matunga (E), Mumbai 400019, India.
authors.forename: Swapnil V
authors.initials: SV
authors.lastname: Pakhale
authors.affiliation: Department of Chemical Engineering, Institute of Chemical Technology, Matunga (E), Mumbai 400019, India. Electronic address: ss.bhagwat@ictmumbai.edu.in.
authors.forename: Sunil S
authors.initials: SS
authors.lastname: Bhagwat
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D012706:Serratia marcescens / Q000201:enzymology*; D014465:Ultrasonics
chemical_list: D010447:Peptide Hydrolases; C022640:serratiopeptidase
keywords: Purification; Serratia marcescens NRRL B 23112; Serratiopeptidase; Three phase partitioning (TPP); Ultrasound assisted three phase partitioning (UATPP)