Abstract
Purification of α-amylase from the cultivation supernatant of recombinant Bacillus subtilis by high-speed counter-current chromatography (HSCCC) in polyethylene glycol (PEG) 4000-inorganic salt aqueous polymer two-phase systems was studied. The effects of sodium chloride concentration on the partition coefficients of α-amylase and total protein were respectively tested in PEG4000-phosphate and PEG4000-citrate aqueous polymer two-phase systems to find the proper range of sodium chloride concentration for the HSCCC purification of α-amylase. α-Amylase was purified from the cultivation supernatant by HSCCC in PEG4000-phosphate system containing 2% (w/w) sodium chloride, yet with considerable loss of activity. PEG4000-citrate aqueous polymer two-phase system containing 2% (w/w) sodium chloride and supplemented with 0.56% (w/w) CaCl2 as protective agent was then successfully applied to purify α-amylase from cultivation supernatant by HSCCC to homogeneity and significantly increased the recovery of α-amylase activity from around 30 to 73.1%.
Original language | English (US) |
---|---|
Pages (from-to) | 215-219 |
Number of pages | 5 |
Journal | Journal of Chromatography A |
Volume | 1070 |
Issue number | 1-2 |
DOIs | |
State | Published - Apr 8 2005 |
Externally published | Yes |
Keywords
- Aqueous polymer two-phase system
- High-speed counter-current chromatography
- Purification
- α-Amylase
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
- Organic Chemistry