The enzyme activity of BnGSTU12
The CDS of BnGSTU12 separated from B. napus cultivar “ZS11” and cloned into the expression vector pET-28a. Approximately a 29 kDa protein band was detected in induced E. coli , which was consistent with the expected protein size. Crude protein was purified using 6*His label and the final concentration reached 1 mg/ml(Figure 1a).
To identify the function of BnGSTU12, recombinant BnGSTU12 was used to measure enzymatic activity toward CDNB substrate. GST enzyme could catalyze the combination of GSH and CDNB as the GSH-conjugation. The enzyme activity increased over time in 2 hours compared with the empty vector (Figure 1b). The results suggested that BnGSTU12 showed GSH-dependent enzyme activity.