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KMID : 0545120140240121690
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Journal of Microbiology and Biotechnology
2014 Volume.24 No. 12 p.1690 ~ p.1698
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Reverse Micellar Extraction of Fungal Glucoamylase Produced in Solid-State Fermentation Culture
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Aliakbar Paraj
Morteza Khanahmadi
Keikhosro Karimi
Mohammad J. Taherzadeh
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Abstract
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Partial purification of glucoamylase from solid-state fermentation culture was, firstly, investigated by reverse micellar extraction (RME). To avoid back extraction problems, the glucoamylase was kept in the original aqueous phase, while the other undesired proteins/ enzymes were moved to the reverse micellar organic phase. The individual and interaction effects of main factors (i.e., pH and NaCl concentration in the aqueous phase, and concentration of sodium bis-2-ethyl-hexyl-sulfosuccinate (AOT) in the organic phase) were studied using response surface methodology. The optimum conditions for the maximum recovery of the enzyme were pH 2.75, 100 mM NaCl, and 200 mM AOT. Furthermore, the optimum organic to aqueous volume ratio (Vorg/Vaq) and appropriate number of sequential extraction stages were 2 and 3, respectively. Finally, 60% of the undesired enzymes including proteases and xylanases were removed from the aqueous phase, while 140% of glucoamylase activity was recovered in the aqueous phase and the purification factor of glucoamylase was found to be 3.0-fold.
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KEYWORD
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Glucoamylase, Solid-State Fermentation, Purification, Reverse Micelle, AOT, Protein
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