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KMID : 1094720090140010060
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Biotechnology and Bioprocess Engineering
2009 Volume.14 No. 1 p.60 ~ p.66
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Optimization of Extraction and Purification of Glucoamylase Produced by Aspergillus awamori in Solid-State Fermentation
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Sangeeta Negi
Banerjee Rintu
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Abstract
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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℃, 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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KEYWORD
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amylase, Aspergillus awamori, enzyme extraction, gel filtration, SDS-PAGE
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