Purification of the threonine-sensitive aspartokinase-homoserine dehydrogenase complex using chromatography on substituted Sepharose columns
| Autor: | William Shive, Weldon M. Harms, Larry Broussard |
|---|---|
| Rok vydání: | 1976 |
| Předmět: |
Threonine
chemistry.chemical_classification Chromatography Chemistry Elution Cyanogen Biophysics Dehydrogenase Cell Biology Chromatography Agarose Biochemistry Sepharose chemistry.chemical_compound Hydrocarbon Multienzyme Complexes Aspartokinase Homoserine Dehydrogenase Yield (chemistry) Escherichia coli Molecular Biology Alkyl |
| Zdroj: | Analytical Biochemistry. 72:16-23 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/0003-2697(76)90501-7 |
| Popis: | The use of cyanogen bromide-activated Sepharose condensed with amines containing alkyl and ω-aminoalkyl groups has provided a series of columns for attempted purifications of the threonine-sensitive aspartokinase-homoserine dehydrogenase of Escherichia coli . Among the alkyl-substituted Sepharose derivatives (Sepharose ∼ NH-(CH 2 ) X H), there was no retardation on the ethyl derivative, slight retardation on the propyl derivative, retardation over a broad band with low yield on the butyl derivative, and strong binding on the pentyl derivative from which active enzyme was not eluted by increased salt concentrations or by a number of other potential eluants. On the ω-aminoalkyl-substituted Sepharose derivatives (Sepharose ∼ NH-(CH 2 ) X -NH 2 ), there was observed no retention on the aminobutyl derivative, retardation with good purification on the aminohexyl and aminoheptyl derivatives, and strong binding on the aminononyl derivative from which active enzyme was not eluted by the several methods attempted. Thus, an appropriate functional group in addition to lengthening the hydrocarbon chain may be necessary for the success of this type of chromatography. |
| Databáze: | OpenAIRE |
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