title: Preparative purification of bromelain (EC 3.4.22.33) from pineapple fruit by high-speed counter-current chromatography using a reverse-micelle solvent system.
abstract: The crude protein of pineapple fruit was purified by high-speed counter-current chromatography (HSCCC). Excellent separation was achieved after careful investigation as follows: cetyltrimethylammonium bromide (CTAB) was selected to prepare the reverse micelle phase at the concentration of 0.10g/mL. The mobile phase was 0.05M sodium phosphate buffer solution, including mobile phase A (pH 9.5, containing 0.2M KCl) used for equilibration and mobile phase B (pH 7.0, containing 0.4M KCl) used for elution. The flow rate of the mobile phase was set at 1.5mL/min and the effluent was monitored at 280nm. Under these conditions, 127.3mg bromelain (EC 3.4.22.33) was produced from 200mg crude sample. A large-scale procedure was then carried out, and 3.01g bromelain was obtained from 5.00g crude extract in a 200min run. The separated protein was analysed by SDS-PAGE, compared with the standard and then identified by TOF/TOF-MS.
authors.affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China.
authors.forename: Lianhong
authors.initials: L
authors.lastname: Yin
authors.affiliation: Institute of Brain Disorders and the Provincial Key Lab for Brain Disorders of Liaoning Province, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China.
authors.forename: C K
authors.initials: CK
authors.lastname: Sun
authors.affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Han
authors.affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Xu
authors.affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China.
authors.forename: Youwei
authors.initials: Y
authors.lastname: Xu
authors.affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Qi
authors.affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshun South Road, Dalian 116044, China. Electronic address: jinyongpeng2005@163.com.
authors.forename: Jinyong
authors.initials: J
authors.lastname: Peng
keywords: Bromelain; CTAB; HSCCC; Protein separation and purification; Reverse micelle