FIG 2.

Purification of recombinant Aly5 (A) and the truncated protein rAly5-T336N (B) from E. coli using Ni²⁺ chelation affinity chromatography. The enzyme purity following each fractionation step was assessed by SDS-PAGE using 13.2% polyacrylamide gels, followed by staining with Coomassie brilliant blue. Lane 1 corresponds to the unstained protein molecular weight marker SM0431 (Thermo), lane 2 corresponds to the induced cell lysate of the E. coli strain containing the control plasmid pET-30a(+), lane 3 corresponds to the induced cell lysate of the E. coli strain containing the recombinant plasmid pE30-Aly5 (A) or pE30-Aly5-T336N (B), lane 4 corresponds to the supernatant fluid of the induced cell lysate, and lane 5 corresponds to the rAly5 (A) or rAly5-T336N (B) purified protein obtained from the supernatant.