Abstract:
p-Glucosidase from African oil bean (Pentaclethra macrophylla) seed was
purified at 4°C by two chromatographic steps. The first step involved Ion-exchange
chromatography using DEAE sephadex A-25 and then gel filtration on sephadex G-l 00.
A 96 fold purification was obtained.
The enzyme was homogeneous as judged by polyacrylamide gel electrophoresis
(PAGE) in the presence ofSDS. The enzyme had a specific activity ofO.96Jlmol/mg and
molar mass of 26KDa. Its optimum activity was observed at pH 5 and 30°C. The enzyme
which was stable at temperatures up to 60°C was gradually inactivated at70°C and above.
The enzyme was stable within the pH range of 3-6 at 27°C. The enzyme was activated by
Zn2+,Mg2+, Ca2+, Na+ and NH/ while Fe3+ and Cu2+strongly inhibited the enzyme
activity. EDT A completely inactivated the enzyme. From Lineweaver-Burk plots, Km
and Vmax values found for D-cellobiose were 5.71 mg/ml and 0.12 umol/ml
respectively.
This work confirms the presence of P-Glucosidase in the African oil bean seed
and it is hoped that further work on this enzyme would help to determine its relevance
industrially.
