Optimization of extraction and purification of glucoamylase produced by Aspergillus awamori in solid-state fermentation |
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| Authors: | Sangeeta Negi Rintu Banerjee |
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| Institution: | (1) Biotechnology Section, Applied Mechanics Department, Motilal Nehru National Institute of Technology, Allahabad, 211004, Uttar Pradesh, India;(2) Agriculture and food Engineering Department, Microbial Biotechnology and Downstream Processing Laboratory, Indian Institute of Technology, Kharagpur, 721302, West Bengal, India |
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| Abstract: | Various parameters such as solvent selection, concentration, soaking time, and temperature were tested in a single bioreactor
in order to determine optimum extraction conditions of glucoamylase, when produced simultaneously with protease by Aspergillus awamari nakazawa MTCC 6652. Optimum conditions were achieved in a 10% glycerol solution soaked for 2 h at 40°C, followed by concentration
of extracted glucoamylase (9,157 U/gds) by acetone precipitation (1:2, v/v), which yielded 51.9% recovery. Ion exchange chromatography
and gel filtration showed specific activities of 270.5 and 337.5 U/mg, respectively, while SDS-PAGE and zymogram analysis
of glucoamylase indicated the presence of three starch-hydrolyzing isoforms with molecular weights of approximately 109.6,
87.1, and 59.4 kDa, respectively |
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| Keywords: | amylase Aspergillus awamori enzyme extraction gel filtration SDS-PAGE |
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