Nimmo, H.G. and Tipton, K.F. (1982) [54] Fructose-bisphosphatase from ox liver. In: Wood, W. A. (ed.) Carbohydrate Metabolism: Part E. Series: Methods in enzymology (90). Academic Press: New York, pp. 330-334. ISBN 9780121819903 (doi: 10.1016/S0076-6879(82)90150-1)
Full text not currently available from Enlighten.
Abstract
This chapter describes the purification and properties of the enzyme fructose-bisphosphatase from ox liver. The enzyme is assayed by coupling the production of fructose 6-phosphate to the reduction of NADP+ in the presence of excess glucosephosphate isomerase and glucose-6-phosphate dehydrogenase. The purification reaction involves extraction, pH treatment, ammonium sulfate fractionation, gradient elution from carboxymethyl (CM)-cellulose, substrate elution from CM-cellulose, and gel filtration on Sephadex G-200. The enzyme shows only one band on sodium dodecyl sulfate (SDS)-polyacrylamide gels and its native molecular weight—as judged by gel filtration—is not altered by the purification, indicating that the purified enzyme is undegraded. The enzyme contains one residue of tryptophan per subunit as judged spectrophotometrically and by amino acid analysis after hydrolysis with methanesulfonic acid. Ox liver fructose-bisphosphatase is stimulated at neutral pH by ethylenediaminetetraacetic acid (EDTA) and other chelating agents; however, EDTA does not bind directly to the enzyme.
| Item Type: | Book Sections |
|---|---|
| Status: | Published |
| Glasgow Author(s) Enlighten ID: | Nimmo, Professor Hugh |
| Authors: | Nimmo, H.G., and Tipton, K.F. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Molecular Biosciences |
| Publisher: | Academic Press |
| ISBN: | 9780121819903 |
University Staff: Request a correction | Enlighten Editors: Update this record
Altmetric
Altmetric