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Efficient hydrolysis of cellulosic biomass into free sugars for the production of bioethanol would allow utilization of an abundant and renewable natural resource. However, complex microcrystalline cellulose in biomass resists digestion. A preferred method to deconstruct cellulose is using enzyme cocktails, but the presence of multiple enzymes with synergistic outcomes for this process complicates direct biochemical analysis as a reliable indicator of deconstruction. Methods to detect release of free sugars rather than direct measurement of enzymatic activity are available but are time-consuming, and do not reflect the production of metabolic inhibitors of microbial growth which may interfere with the subsequent fermentation of sugars to ethanol. The work presented here describes an automated method for detecting the release of free sugars from enzymatic digestion of cellulosic biomass that allows for real-time measurements during the enzyme treatments and provides an indication of metabolic inhibitors that can interfere with subsequent growth of microbes during fermentation.


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