Purification and Characterization of ?-N-Acetylgalactosaminidase from <i>Hilsha ilisha</i>

Authors

  • M. H. Rashid Department of Pharmacy World University of Bangladesh, Dhanmodi, Dhaka-1205, Bangladesh
  • A. H. M. K. Alam Department of Pharmacy University of Rajshahi, Rajshahi-6205, Bangladesh
  • J. Uddin Department of Pharmacy World University of Bangladesh, Dhanmodi, Dhaka-1205, Bangladesh
  • N. Karim Department of Pharmacy Jahangirnagar University, Savar, Dhaka, Bangladesh
  • M. G. Sadik Professor, Department of Pharmacy, Rajshahi University, Rajshahi- 6205, Bangladesh

DOI:

https://doi.org/10.3329/jsr.v9i2.30525

Keywords:

Hilsha ilisha, ?-N-Acetylgalactosaminidase, ?-Galactosidase, Purification.

Abstract

The objective of this study was to purify and characterize the alpha-N-acetylgalactosaminidase (?-GalNAcase) from hilsha fish, Hilsha ilisha. Digestive organ of hilsha fish was found to contain a large amount of ?-GalNAcase in compared with the other tissues examined. ?-GalNAcase was purified from the crude extract of hilsha fish by ammonium sulphate precipitation, Sephadex G-200 gel filtration, and SP-Sephadex C-50 column chromatography. The purified enzyme gave a single band on sodium dodecylsulphate-polyacrylamide gel electrophoresis and exhibited a molecular mass of 48 kDa. The final preparation of ?-GalNAcase showed 3.02% ?-galactosidase activity. The enzyme had an optimum pH of 4.0 and was found to be quiet heat stable at 37°C. Inhibition studies with metal ions demonstrated that the enzyme was highly inhibited by silver and mercury ions. Both N-acetylgalactosamine and galactose affect the enzyme activity. Kinetic studies with the enzyme showed that the KM value for p-nitrophenyl-?-N-acetylgalactosaminide substrate was 3.31 mM and the Vmax value was 35.04 unit/mg.

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Author Biography

M. G. Sadik, Professor, Department of Pharmacy, Rajshahi University, Rajshahi- 6205, Bangladesh

Professor, Department of Pharmacy, Rajshahi University, Rajshahi- 6205, Bangladesh

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Published

2017-04-20

How to Cite

Rashid, M. H., Alam, A. H. M. K., Uddin, J., Karim, N., & Sadik, M. G. (2017). Purification and Characterization of ?-N-Acetylgalactosaminidase from <i>Hilsha ilisha</i>. Journal of Scientific Research, 9(2), 235–246. https://doi.org/10.3329/jsr.v9i2.30525

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Section

Section B: Chemical and Biological Sciences