Individual enzymes are cloned on replicating plasmids in C. thermocellum and purified with his-tags. (IMAGE)
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Individual enzymes are cloned on replicating plasmids in C. thermocellum and purified with his-tags. Kinetic parameters are determined by enzyme assays where consumption or production of one metabolite (usually NAD(P)(H)) is measured continuously. For cell lysates, various substrates are gradually fed into the system, metabolite concentrations are measured at discrete time points using HPLC and LC/MS to measure over 40 metabolites.
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Credit Daniel Olson/Dartmouth Engineering
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Credit Daniel Olson/Dartmouth Engineering
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