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Analysis of mandelonitrile lyase and β‐glucosidase from sweet almonds by combined electrophoretic techniques
Author(s) -
Chiari Marcella,
Gelain Arianna,
Riva Serigo,
Tura Debora
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150181129
Subject(s) - electrophoresis , chemistry , lyase , biochemistry , chromatography , enzyme
Almonds are a rich source of mandelonitrile lyase (oxynitrilase) and β‐glucosidase. The isolation of these two enzymes from sweet almonds reqires 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, β‐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.