title: Purification of serine hydroxymethyltransferase from Bacillus stearothermophilus with ion-exchange high-performance liquid chromatography.
abstract: The gene of serine hydroxymethyltransferase (SHMT) of a thermophilic bacterium Bacillus stearothermophilus was expressed in Escherichia coli, and SHMT was successfully purified from the crude extract of E. coli in two steps while maintaining the enzymatic activity. The purification steps involved ammonium sulphate precipitation followed by high-performance liquid chromatographic separation using the anion-exchange column Fractogel EMD DEAE-650(S). In addition to the DEAE column, three other types of anion- and cation-exchange columns were also studied for their ability to separate SHMT, and the performance of the four columns were compared.
authors.affiliation: Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ide
authors.forename: K
authors.initials: K
authors.lastname: Hamaguchi
authors.forename: S
authors.initials: S
authors.lastname: Kobata
authors.forename: A
authors.initials: A
authors.lastname: Murakami
authors.forename: Y
authors.initials: Y
authors.lastname: Kimura
authors.forename: K
authors.initials: K
authors.lastname: Makino
authors.forename: M
authors.initials: M
authors.lastname: Kamáda
authors.forename: S
authors.initials: S
authors.lastname: Miyamoto
authors.forename: T
authors.initials: T
authors.lastname: Nagaya
authors.forename: K
authors.initials: K
authors.lastname: Kamogawa
mesh_terms: D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D012696:Glycine Hydroxymethyltransferase / Q000302:isolation & purification*; D014169:Transformation, Bacterial
chemical_list: D012696:Glycine Hydroxymethyltransferase; D000645:Ammonium Sulfate