Partial purification and characterisation of α-amylases from the digestive tract of the Indian major carp Labeo rohita (Hamilton, 1822)


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Authors

  • Umalatha . Regional Research Center of Central Institute of Freshwater Aquaculture Hesserghatta Lake Post, Bangalore-560 089
  • N. Sridhar
  • Jairam Prasad Kushwaha
  • Vadlapudi Kumar

https://doi.org/10.21077/ijf.2019.67.2.59187-09

Abstract

Partial purification of α-amylases from the digestive tract of the Indian major carp Labeo rohita (Hamilton, 1822) through acetone fractionation and ion exchange chromatography (DEAE-SephadexA-50) resulted in 8-fold purification with 86% recovery. Characterisation of amylase activity revealed two pH optima at 4.5 and 6.5. Activity was stable over wide pH ranges of 3.5 to 4.5 and 7 to 12. Optimum incubation temperature was 35°C. The enzyme lost 91% activity at 60°C within 15 min and was inhibited by Amylase inhibitor Type-1 (wheat); 1, 10 Phenanthroline, Ethylene diamine tetra-acetate (EDTA) and Phenyl methyl sulphonyl fluoride (PMSF). Heavy metal ions Hg++ and Cu++ strongly inhibited the enzyme activity, while Zn++ and Bi++ inhibited to a lesser extent. Native polyacrylamide gel electrophoresis of the purified amylase fractions revealed four bands, with corresponding molecular weights of 43.59, 52.36, 55.42 and 54.01 kDa. α-Amylase activity from L. rohita exhibited linear hydrolysis of starch upto 7% concentration in 60 min.

Keywords: α-Amylase, ,Characterisation, Digestive tract, Labeorohita, Purification

Author Biography

  • Umalatha ., Regional Research Center of Central Institute of Freshwater Aquaculture Hesserghatta Lake Post, Bangalore-560 089

    RRC of CIFA,

    Principal Scientist

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Submitted

2016-06-15

Published

2020-06-30

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Section

Articles

How to Cite

., U., Sridhar, N., Kushwaha, J. P., & Kumar, V. (2020). Partial purification and characterisation of α-amylases from the digestive tract of the Indian major carp Labeo rohita (Hamilton, 1822). Indian Journal of Fisheries, 67(2). https://doi.org/10.21077/ijf.2019.67.2.59187-09