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The isolation of an aliphatic amidase from Pseudomonas aeruginosa
Author(s) -
Lilly M. D.,
Clarke P. H.,
Houldsworth M.,
Currie J. A.,
Dunnill P.
Publication year - 1969
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260110303
Subject(s) - amidase , fractionation , ammonium sulfate , chromatography , polyethylene glycol , chemistry , ammonium , ammonium sulfate precipitation , isolation (microbiology) , pseudomonadaceae , enzyme , pseudomonas aeruginosa , biochemistry , organic chemistry , bacteria , biology , microbiology and biotechnology , genetics , size exclusion chromatography
A pilot‐scale process for the isolation of an aliphatic, amidase from Pseudomonas aeruginosa has been developed. A constitutive, partially irrepressible mutant was employed to give a high initial enzyme concentration. An existing laboratory isolation procedure has been scaled up and modified particularly by substitution of polyethylene glycol for ammonium sulfate precipitation as the first stage in the conversion of the fractionation to continuous operation. Full recovery of activity was achieved with the modification. The recovery of enzyme from a subsequent chromatographic stage was 85% and the maximum overall purification was 28‐fold.