PURIFICATION AND CHARACTERIZATION OF a-GLUCOSIDASE FROM MOSS HYOPHILLA NYMANIANA (FLEISH.) MENZEL
Objective: The present study was undertaken to extract and purify Î±-glucosidase N-linked glycosylation enzyme from moss Hyophilla nymaniana
Methods: Frozen protonemal cells were taken for crude enzyme extraction, and the enzyme Î±-glucosidase was purified from the prepared crude
enzyme extract by ammonium sulfate (NH
precipitation, gel filtration and finally on diethylaminoethyl sephadex column chromatography.
Results: The final purification step of the enzyme resulted in 35 fold purification with a recovery of 4%. A single protein band of 72Â±5 kilodalton was
seen on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The physiochemical characterization of the enzyme reveled the enzyme had
a wide pH stability range 4-7 with optimum pH 5 while the temperature stability study revealed the enzyme was stable up to 60Â°C while the optimum
temperature of the purified enzyme was 45Â°C. The enzyme was strongly inhibited by Hg
at 1 mM concentration while Mg
enhanced the enzyme activity at the same concentration. The kinetic study of the enzyme showed K
of the enzyme 5.2 mM/ml and 8.6 U/ml,
Conclusion: The wide pH and temperature stability range show its suitability toward industrial application.
Keywords: Î±-glucosidase, Hyophilla nymaniana (Fleish.) Menzel. gel filtration, Diethylaminoethyl sephadex column chromatography.
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