PURIFICATION AND CHARACTERIZATION OF a-GLUCOSIDASE FROM MOSS HYOPHILLA NYMANIANA (FLEISH.) MENZEL

  • Ramesh Chandra Birla Institute of Technology, Mesra, Ranchi, India
  • Rashmi Mishra Birla Institute of Technology, Mesra, Ranchi, India

Abstract

Objective: The present study was undertaken to extract and purify α-glucosidase N-linked glycosylation enzyme from moss Hyophilla nymaniana
(Fleish.) Menzel.
Methods: Frozen protonemal cells were taken for crude enzyme extraction, and the enzyme α-glucosidase was purified from the prepared crude
enzyme extract by ammonium sulfate (NH
4
)
2
SO
4
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 72±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 60°C while the optimum
temperature of the purified enzyme was 45°C. The enzyme was strongly inhibited by Hg
2+
and Ag
2+
at 1 mM concentration while Mg
ions
enhanced the enzyme activity at the same concentration. The kinetic study of the enzyme showed K
m
and V
of the enzyme 5.2 mM/ml and 8.6 U/ml,
respectively.
Conclusion: The wide pH and temperature stability range show its suitability toward industrial application.
max
Keywords: α-glucosidase, Hyophilla nymaniana (Fleish.) Menzel. gel filtration, Diethylaminoethyl sephadex column chromatography.

Author Biographies

Ramesh Chandra, Birla Institute of Technology, Mesra, Ranchi, India
Department of Bio-Engineering, Associate Professor
Rashmi Mishra, Birla Institute of Technology, Mesra, Ranchi, India
Department of Bio-Engineering, Research scholar

References

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How to Cite
Chandra, R., and R. Mishra. “PURIFICATION AND CHARACTERIZATION OF a-GLUCOSIDASE FROM MOSS HYOPHILLA NYMANIANA (FLEISH.) MENZEL”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 9, no. 1, Jan. 2016, pp. 179-82, https://innovareacademics.in/journals/index.php/ajpcr/article/view/9674.
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Original Article(s)