Background: 5'-Phosphodiesterase (5'-PDE) is an enzyme that hydrolyzes RNA to form 5'-inosine monophosphate (5'-IMP) and 5'-guanosine monophosphate (5'-GMP). These 5'-nucleotides can function as flavor enhancers. Adzuki beans (Vigna angularis L.) are found to be high in 5'-PDE.
Methods: 5'-phosphodiesterase (5'-PDE) enzyme was characterized from adzuki beans, in which the optimum pH and temperature were determined. In addition, the stability of 5'-PDE was assessed at different pH and temperature. The effects of cations and EDTA were evaluated to characterize the 5'-PDE enzymes further.
Results: The alkaline 5'-phosphodiesterase has an optimum pH of 8.5. This enzyme is also thermostable, with an optimum temperature of 80°C. The stability in terms of temperature and pH was also determined, and was found to be stable in the pH range of 7.0-8.5. This enzyme was found to retain more than 80% of its activity for 4 days at 60 and 65°C. In addition, the effects of 14 different metal ions, 4 types of detergents and ethylenediaminetetraacetic acid (EDTA) on 5'-PDE were studied. Ca2+, K+, Mg2+ and Li+ activated 5'-PDE while Na+, Zn2+, Ni+, Hg+, Cu2+, Pb2+, Fe2+, Al3+, Ba2+ and Co2+ were inhibitory. EDTA, Triton X-100 and sodium dodecyl sulfate (SDS) were strong inhibitors of 5'-PDE, while Tween 80 and Tween 20 were slightly inhibitory. The effects of cations and EDTA suggest that 5'-PDE from adzuki beans is a metalloenzyme.
Conclusions: Although 5'-PDE from adzuki beans has a high temperature optimum of 80°C, the enzyme is more stable at 60°C, and different cations affected the activity of the enzyme differently.
Keywords: 5’-phosphodiesterase; cations; detergents; optimum pH; thermostability.