Selection of Conditions for Cellulase and Xylanase Extraction from Switchgrass Colonized by <Emphasis Type="Italic">Acidothermus cellulolyticus</Emphasis> |
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Authors: | Farzaneh Rezaei Lawrence D Joh Hiroyuki Kashima Amitha P Reddy Jean S VanderGheynst |
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Affiliation: | (1) Department of Biological and Agricultural Engineering, University of California, One Shields Avenue, Davis, CA 95616, USA;(2) Joint BioEnergy Institute, Emeryville, CA 94608, USA; |
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Abstract: | Solid-state fermentation has been widely used for enzyme production. However, secreted enzymes often bind to the solid substrate
preventing their detection and recovery. A series of screening studies was performed to examine the role of extraction buffer
composition including NaCl, ethylene glycol, sodium acetate buffer, and Tween 80, on xylanase and cellulase recovery from
switchgrass. Our results indicated that the selection of an extraction buffer is highly dependent on the nature and source
of the enzyme being extracted. While a buffer containing 50 mM sodium acetate at pH 5 was found to have a positive effect
on the recovery of commercial fungal-derived cellulase and xylanase amended to switchgrass, the same buffer had a significant
negative effect on enzyme extraction from solid fermentation samples colonized by the bacterium Acidothermus cellulolyticus. Xylanase activity was more affected by components in the extraction buffers compared to cellulase. This study demonstrated
that extraction followed by diafiltration is important for assessing enzyme recovery from solid fermentation samples. Reduction
in activity due to compounds present in the switchgrass extracts is reversible when the compounds are removed via diafiltration. |
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