E. coli penicillin acylase isolation by selective release, aqueous two-phase partitioning and ultrafiltration.
A procedure has been developed for the purification of penicillin acylase from E. coli fermentation broths. The method described is based primarily on selective release from biomass, charge-directed partitioning in an aqueous two-phase system, and the use of ultrafiltration membranes to recycle the modified polyethylene glycol required and to further purify the enzyme. Each step has been discussed and comparisons have been made with other approaches, where possible. Suggestions have been made for optimisation in process engineering. The approach developed may be applicable to other beta-lactam antibiotic acylases or more generally to some of the other E. coli periplasmic proteins.
Guan
Y
Y
Department of Molecular Biology and Biotechnology, The University, Sheffield, UK.
Treffry
T E
TE
Lilley
T H
TH
D001681:Biological Assay; D002458:Cell Fractionation; D004926:Escherichia coli / Q000201:enzymology*; D007218:Industrial Microbiology / Q000379:methods*; D008956:Models, Chemical; D010399:Penicillin Amidase / Q000302:isolation & purification*; D010865:Pilot Projects; D012995:Solubility; D014462:Ultrafiltration
D010399:Penicillin Amidase