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Purification and Concentration of Alkaline Phosphatase by Selective Permeabilization of Escherichia c oli Using Reverse Micellar Solutions
Author(s) -
BansalMutalik Ritu,
Gaikar Vilas G.
Publication year - 2003
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp034141r
Subject(s) - chemistry , escherichia coli , alkaline phosphatase , periplasmic space , chromatography , nucleic acid , solvent , aqueous solution , micellar solutions , aqueous two phase system , micelle , enzyme , biochemistry , organic chemistry , gene
Abstract 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.