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Biotechnology and Bioprocess Engineering

, Volume 14, Issue 1, pp 60–66 | Cite as

Optimization of extraction and purification of glucoamylase produced by Aspergillus awamori in solid-state fermentation

  • Sangeeta NegiEmail author
  • Rintu Banerjee
Article

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

Keywords

amylase Aspergillus awamori enzyme extraction gel filtration SDS-PAGE 

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Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg GmbH 2009

Authors and Affiliations

  1. 1.Biotechnology Section, Applied Mechanics DepartmentMotilal Nehru National Institute of TechnologyAllahabadIndia
  2. 2.Agriculture and food Engineering Department, Microbial Biotechnology and Downstream Processing LaboratoryIndian Institute of TechnologyKharagpurIndia

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