(R)-mevalonate → (+-)-Mevalonolactone

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GC-MS analysis for in vivo mevalonate production. As for screening the single mutation library, a single colony harboring pBADMevT (wild type tHMGR or its mutant variants) was inoculated into LB medium containing Cm50 and grown overnight at 37° C. An aliquot (50 μl) of this seed culture was inoculated into fresh LB medium (5 ml) containing Cm50 and 13.3 mM (+)-L-arabinose, and grown for 24 hours at 37° C. An aliquot of culture (560 μl) was mixed with 140 μl of 0.5 M HC1 to dehydrate the mevalonate to form mevalonolactone, and 700 μl of ethyl acetate was then added to the sample. The mixture was vortexed for 5 minutes, and the ethyl acetate was analyzed by GC-MS using a GC oven temperature program of 90° C. for 1 min, then ramping 30° C./min to 250° C. for CyclosilB capillary column analysis. Mevalonolactone was identified from its mass spectra and retention time by comparison to an authentic standard.

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