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EC Number Application Commentary Reference
Display the reaction diagram Show all sequences 4.2.3.77analysis use of NMR spectroscopy as a tool to analyze the kinetics of enzyme reactions using progress curves. The protocol presented is also a simple and direct approach for the measurement of enzyme kinetics in the presence of synthetic inhibitors. The conversion of farnesyl diphosphate into (+)-germacrene D by the enzyme germacrene D synthase involves an amphiphilic substrate forming micelles and a water insoluble product. Using proper controls, the conversion can well be analyzed by the progress curve approach using the Lambert W function 729283
Display the reaction diagram Show all sequences 4.2.3.77synthesis improved conditions for expression of Zingiber officinale (+)-germacrene synthase in Escherichia coli. Comparison of bacterial strains; BL21 (DE3), BL21 (DE3) Tuner BL21(DE3) pLysS and BL21 (DE3) pLysS Tuner using different inducing agents. The change between BL21 (DE3) cells and BL21 (DE3) Tuner, induced with IPTG, leads to a twofold increase in enzyme activity in the soluble fraction while a reduction in activity is observed when using the pLysS strains. The same doubling of activity is observed for germacrene synthase when the commonly used BL.21 (DE3) is induced with The Inducer. Addition of 2.5 mM glycine betaine and 660 mM sorbitol to the bacterial growth media results in reduction of growth rate and biomass yield but under these conditions the best overall protein production is obtained 682659
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