Purification and concentration of alkaline phosphatase by selective permeabilization of Escherichia coli using reverse micellar solutions.
Recovery of alkaline phosphatase (AP) from the periplasm of Escherichia coli using reverse micellar solutions (RMSs) of sodium dioctyl sulfosuccinate (AOT) in aliphatic hydrocarbons has been attempted. A variety of surface-active agents, solvents, and reverse micellar conditions were screened, and an excellent recovery of the enzyme in a concentrated form, with a high purification factor, was obtained in a single-step process. The permeabilization process strongly depended on the water content of the RMS as well as on the amount of water coating the microbial cell surface. The product was almost free from nucleic acids. In addition, because of the low affinity of AOT and the organic solvent for the aqueous phase, contamination by the permeabilizing agents would also be negligible.
Bansal-Mutalik
Ritu
R
Institute of Chemical Technology, University of Mumbai, Matunga, Mumbai-400 019, India.
Gaikar
Vilas G
VG
D000469:Alkaline Phosphatase / Q000096:biosynthesis* / Q000302:isolation & purification*; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology*; D003102:Colloids / Q000494:pharmacology*; D004143:Dioctyl Sulfosuccinic Acid / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology*; D008823:Micelles
D003102:Colloids; D008823:Micelles; D004143:Dioctyl Sulfosuccinic Acid; D000469:Alkaline Phosphatase