Farnesyl diphosphate synthase (FPPS) is an important enzyme for the formation of fifteen-carbon terpenes, also known as sesquiterpenes. Many FPPS structures are known, but the structure of the homolog from the thermophilic archeon <i>Thermoplasma volcanium </i>is not known. Studying the structural adaptations of this enzyme that allow it thrive in high temperatures should yield insights into its activity. It is hoped that the expression, purification, and crystallization of FPPS will enable to determination of the difference in this enzyme that allows for the thermophilic stability. The FPPS gene was cloned into the plasmid, pGEX2T, which provided a N-terminal GST tag. BL21DE3 cells were induced to produce the protein, and purification was attempted with reduced glutathione resin. Expression, however, was limited due to insolubility caused by the protein misfolding. The GST-FPPS plasmid was transformed into a new cell line, Lemo21DE3, and successfully expressed.

Expression and purification of farnesyl diphosphate synthase from <i>Thermoplasma volcanium</i>

249th National Meeting (2015)

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Expression and purification of farnesyl diphosphate synthase from Thermoplasma volcanium