Rapid expression of pyruvate decarboxylase from Zymomonas mobilis in E. coli BL21 LysY/I ^q

E. coli BL21 DE3 strain is commonly used to express and purify non-toxic prokaryotic proteins in high yields. Traditionally, IPTG based induction of the host strain at reduced temperatures for extended period (12-16 h) is performed to obtain high yield of functional proteins. It is desirable to explore methods which result in high yield of protein within a short period of time. We report rapid purification of pyruvate decarboxylase (PDC) enzyme from Zymomonas mobilis using E. coli BL21 pLysY/I ^q as a host strain. High yield of purified PDC at 0.33 (±0.02) mg was obtained after two-hour induction by 0.6 mM IPTG at 37 ^° C. The enzyme yield was comparable to 0.37 (±0.08) mg obtained in E. coli BL21 DE3 strain (used as control) after 16 h induction by 0.6 mM IPTG at 18 ^° C. Similar values of the maximum specific activity of the enzyme expressed and purified at 37 ^° C and 18 ^° C were obtained at 78.31 (±1.13) in strain LysY/I ^q and 85.73 (±4.39) µmol/min/mg protein in strain DE3, respectively. In almost all IPTG treatments, the kinetic parameters of the purified enzyme - app K _m , app V _max , K _cat and K _cat /K _m -also did not vary remarkably between the two temperature regimes. Based upon the data presented here, we propose that E. coli BL21 LysY/I ^q strain has potential to serve as a host for efficient and rapid expression (2 h) of non-toxic proteins. Results of this study will aid in cell free system study which require rapid scale up of the complexity of metabolic pathways by utilizing multiple purified enzymes involved in bioconversion of the substrate of interest.