Analysis of mandelonitrile lyase and beta-glucosidase from sweet almonds by combined electrophoretic techniques

Electrophoresis
M ChiariD Tura

Abstract

Almonds are a rich source of mandelonitrile lyase (oxynitrilase) and beta-glucosidase. The isolation of these two enzymes from sweet almonds requires fractional ammonium sulfate precipitation followed by ion-exchange chromatography on diethylaminoethyl-(DEAE) and carboxymethylcellulose (CMC) columns. In the present investigation different electrophoretic techniques such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing in immobilized pH gradients (IEF-IPG), and capillary electrophoresis were used to characterize these two enzymes. For the first time, beta-glucosidase and oxynitrilase were separated in an immobilized pH gradient of one pH unit. Capillary zone electrophoresis (CZE) was an excellent tool for analysis of the purity of enzyme preparations, achieving complete separation of various protein constituents in only 15 min. CZE showed a resolving capacity for the separation of enzyme forms comparable to that of isoelectric focusing in an immobilized pH gradient.

References

Apr 15, 1978·Archives of Biochemistry and Biophysics·D E Walker, B Axelrod
Sep 25, 1974·Journal of Chromatography·P G Righetti, J W Drysdale
Mar 1, 1972·Hoppe-Seyler's Zeitschrift für physiologische Chemie·E Gerstner, E Pfeil
Feb 12, 1981·Biochemical and Biophysical Research Communications·L KissP Nánási

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Citations

Jun 1, 2013·Applied Biochemistry and Biotechnology·Ming-Zhu GuBang-Ce Ye
May 22, 2010·Biomacromolecules·Hans Bäumler, Radostina Georgieva

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