Human PTRHD1 was successfully overexpressed as a soluble Nterminal
hexahistidine fusion protein in E. coli. Improved solubility,
leading to higher overall yields, was found for expression in M9
minimal media over richer LB media. Similarly, starting from
scraped plates as opposed to liquid cultures, expression at 14 C,
and a media to flask ratio of 1:4 resulted in more soluble PTRHD1
and the greatest overall yield. To further maximize yield, variable
IPTG concentrations and induction durations were tested. IPTG
concentration had the largest impact with 250 mM IPTG producing