PMID: 5145May 13, 1976

Alpha-D-Mannosidase. Preparation and properties of free and insolubilized enzyme

Biochimica Et Biophysica Acta
V Sheperd, R Montgomery

Abstract

Alpha-D-Mannosidase (alpha-D-mannoside mannohydrolase, EC 3.2.1.24) has been purified to homogeneity as demonstrated by polyacrylamide gel electrophoresis and ultracentrifugation. The molecular weight of the enzyme is approx. 200000; the protein appears to contain 4 subunits, with molecular weights of 66000 and 44000. The enzyme was immobilized on Sepharose and the properties of the coupled and free enzyme were compared. Both were stable up to 70 degrees C with rapid loss of activity between 75-80 degrees C; both retained 25-30% activity in 6 M urea and 65% of the original activity could be restored in the coupled preparation by removal of the urea. The pH maximum of each form was approximately the same, with the maximum of the immobilized enzyme shifted slightly to a lower pH. The coupled alpha-D-mannosidase presented in this report offers the possibility of digesting high molecular weight substrates, such as glycoproteins, with the advantages of (1) recovering large quantities of digested substrate; (2) recovery of the active glycosidase; and (3) digestion at high temperatures and under conditions that denature many proteins.

References

Apr 15, 1975·FEBS Letters·D RobinsonB Winchester
Jul 1, 1969·Analytical Biochemistry·R M ZachariusJ J Woodlock
Jun 1, 1973·Archives of Biochemistry and Biophysics·H B Bull, K Breese
Oct 1, 1971·Analytical Biochemistry·Y C LeeJ Scocca
Nov 1, 1968·Archives of Biochemistry and Biophysics·T J Langley, F R Jevons
Mar 1, 1969·Archives of Biochemistry and Biophysics·S Panyim, R Chalkley
Aug 19, 2007·Nucleic Acids Research·Yuan YuanMaurice S Swanson

Citations

Sep 28, 1984·Biochemical and Biophysical Research Communications·R MathurA S Balasubramanian
Feb 15, 1983·European Journal of Biochemistry·D J BowlesS E Marcus

Related Concepts

Disaccharidases
Hot Temperature
Hydrogen-Ion Concentration
Mannosidase
Ovalbumin
Plasma Protein Binding Capacity
Sepharose
Carmol
Macromolecular Compounds

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