A rapid and sensitive spectrophotometric assay for prunasin hydrolase activity employing purified mandelonitrile lyase

✍ Scribed by Mara Gross; Gregory H. Jacobs; Jonathan E. Poulton

• Book ID: 102985924
• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 593 KB
• Volume: 119
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90660-1

✦ Synopsis

A rapid, sensitive, and inexpensive assay for prunasin hydrolase activity is described. Prunasin hydrolase is incubated with saturating concentrations of prunasin in the presence of excess purified mandelonitrile lyase. Under these conditions, the mandelonitrile formed by prunasin hydrolysis is immediately converted to benzaldehyde, which may be determined spectrophotometrically at 249.6 nm. The rate of prunasin hydrolysis is determined from the rate of change of absorbance at this wavelength. A single ion-exchange chromatographic procedure is also described which allows the facile purification of mandelonitrile lyase to near homogeneity with greater than 70% recovery.