*2.2. Recombinant Protein Expression*

The pET-14b cDNA construct [2] was transformed into one-shot *E. coli* BL21(DE3) pLysS competent cells (Invitrogen), plated on LB agar medium containing ampicillin (100 μg/mL) (Sigma Aldrich), and chloramphenicol (34 μg/mL) (Sigma Aldrich). One colony was then incubated in 10 mL of LB broth (with antibiotics) and allowed to grow overnight at 37 ◦C under orbital agitation. Then, this pre-culture was expanded into 1L of LB broth with antibiotics and allowed to grow at 37 ◦C until the OD at 550 nm reached 0.5. For the induction of heterologous protein expression, isopropyl-d-thiogalactoside (IPTG, ThermoFisher Scientific, Waltham, MA, USA) was added at a final concentration of 0.1 mM, and induction of the culture was performed for 4 h at 23 ◦C.
