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

Issue

Vol. 67 No. 2 (2020)

Section

Articles

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The copyright of the articles published in Indian Journal of Fisheries vests with the Indian Council of Agricultural Research, who has the right to enter into any agreement with any organization in India or abroad engaged in reprography, photocopying, storage and dissemination of information contained in these journals. The Council has no objection in using the material, provided the information is being utilized for academic purpose but not for commercial use. Due credit line should be given to the ICAR where information will be utilized.

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