Abstract
An enzyme having activity toward n-hexanol was purified from apple, and its biochemical characteristics were analyzed. The purification steps consisted of sedimentation with ammonium sulfate, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephadex G-100 column. The obtained enzyme had a yield of 16.00% with a specific activity of 18879.20 U/mg protein and overall purification of 142.77-fold. The enzyme showed activity to isoamylol, 1-propanol, n-hexanol, and isobutanol but not toward methanol and ethanol. With n-hexanol as a substrate, the optimum conditions were pH 4.0 and 30 °C for enzyme activity and pH 3.0-4.0 and temperatures below 40 °C for enzyme stability. The enzyme activity was increased significantly by adding l-cysteine and Fe 2+ at all tested concentrations and slightly by Zn 2+ at a high concentration but decreased by additions of EDTA, Ga 2+, K +, Mg 2+, sodium dodecyl sulfate (SDS), sodium aluminum sulfate (SAS), dithiothreitol (DTT), and glutathione (GSH). The enzyme activities toward n-hexanol and n-hexanal were increased by NADH but decreased by NAD +, in contrast to a decrease toward n-hexane by addition of both NAD + and NADH.
Original language | English |
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Pages (from-to) | 3246-3252 |
Number of pages | 7 |
Journal | Journal of Agricultural and Food Chemistry |
Volume | 60 |
Issue number | 12 |
DOIs | |
State | Published - 28 Mar 2012 |
Externally published | Yes |
Keywords
- apple
- characteristics
- enzyme
- higher alcohol
- kinetic
- n-hexanol
- purification