(R)-mevalonate → (+-)-Mevalonolactone

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.

1. additioncontaining Cm50
2. waitgrown for 24 hours at 37° C
3. additionAn aliquot of culture (560 μl) was mixed with 140 μl of 0.5 M

ORD-Eintrag

ord-1d1267ad8b0e44aca62028255b3c1832

Quelldatensatz

uspto-grants-2012_04

DOI

10.6084/m9.figshare.5104873.v1

Patentnummer

US08158383B2