Open AccessCitrate synthases from the Archaea: Development of a bio‐specific, affinity chromatography purification procedure
Author(s) -
James Keith D.,
Russell Rupert J.M.,
Parker Lynne,
Daniel Roy M.,
Hough David W.,
Danson Michael J.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb06886.x
Subject(s) - archaea , thermophile , biochemistry , halophile , affinity chromatography , citrate synthase , elution , biology , chromatography , chemistry , bacteria , gene , enzyme , genetics
Citrate synthases from both thermophilic and halophilic Archaea have been purified to homogeneity using affinity chromatography on Matrex Gel Red A and elution with a combination of substrate (oxaloacetate) and product (coenzyme A). In a number of cases, purification from cell‐extract to protein suitable for N‐terminal sequencing can be achieved by this single‐step procedure. The method is particularly useful in the rapid purification of a thermophilic archaeal citrate synthase from a cloned gene expressed in a mesophilic host.