Hyper-activation of foldase-dependent lipase with lipase-specific foldase

The LST-03 lipase from Pseudomonas aeruginosa LST-03 requires lipase-specific foldase for activation. Abundant expression of the active lipase was successfully accomplished with individual expression of the lipase and foldase in a heterologous host and subsequent in vitro activation. Although the activity of the native lipase from culture supernatant of P. aeruginosa LST-03 was 110 kI.U./g, that after in vitro activation using individually expressed lipase and foldase was 228 kI.U./g. Furthermore, the activity after in vitro activation with afterwards adding calcium ions was 359 kI.U./g. However, the incubation of the lipase with the foldase in the presence of calcium ions resulted in a small conformational transition and low activation levels of the lipase by the foldase. The lipase showed high affinity for the foldase in the presence of calcium ions. The results indicate that in a cellular environment that contains calcium ions, the lipase would not become a hyperactive form by the foldase.