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.