Partial purification and biochemical characterization of alkaline 5'-phosphodiesterase from barley malt sprouts |
| |
Authors: | Beluhan S Karmeli? I Novak S Mari? V |
| |
Institution: | (1) Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6/IV, 10000 Zagreb, Croatia |
| |
Abstract: | An alkaline 5-phosphodiesterase (5-PDE) from barley (Hordeum distichum) malt sprouts was partially purified by thermal treatment and acetone precipitation to diminish phosphomonoesterase (PME) activity. 5-PDE was purified 40-fold to a specific activity of 30 U mg–1 protein with a final yield of about 32%. With synthetic substrate, the enzyme had an optimum pH of 8.9, maximum activity at 70 °C over 10 min, and a Km of 0.26 mM. The partially purified enzyme was activated by 10 mM Mg2+ up to 168% of the original activity, while Zn2+, Mn2+ and Cu2+ ions, chelating agent (EDTA) and NaN3 (1–10 mM), and 5-ribonucleotides (1–5 mM) were inhibitory. Final enzyme preparation was stable over 8 d at 4 °C), at 70 °C for up to 120 min and without loss of activity over 90 d at –18 °C. |
| |
Keywords: | alkaline 5-phosphodiesterase" target="_blank">gif" alt="prime" align="BASELINE" BORDER="0">-phosphodiesterase barley malt sprouts 5-ribonucleotides" target="_blank">gif" alt="prime" align="BASELINE" BORDER="0">-ribonucleotides storage stability thermostability |
本文献已被 PubMed SpringerLink 等数据库收录! |
|