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Figure 6 | Microbial Cell Factories

Figure 6

From: Enhancement of solubility in Escherichia coli and purification of an aminotransferase from Sphingopyxis sp. MTA144 for deamination of hydrolyzed fumonisin B1

Figure 6

Enterokinase cleavage and aminotransferase activity of purified MBP-FumI fusion protein. (A) Total (T), soluble (S) and insoluble (I) fractions of purified MBP-FumI fusion protein before addition of recombinant enterokinase (0 h rEK), after 6 h and after 24 h incubation with recombinant enterokinase were subjected to SDS-PAGE (10% polyacrylamide, GelCode Blue staining). Lane M, molecular mass marker. The molecular masses of MBP (42.6 kDa), FumI (46.9 kDa) and the fusion protein (89.5 kDa) are indicated. (B) Enzyme activity of purified MBP-FumI fusion protein after incubation with (+ rEK) or without (Ø rEK) recombinant enterokinase. For comparison, 6xHis-tagged aminotransferase (FumI-HIS) purified from E. coli ArcticExpress(DE3) by affinity chromatography was included. Protein concentration was 20 μg/ml for each sample. Samples were incubated with 15 μM HFB1 in 20 mM Tris-HCl buffer (pH 8.0) at 25°C, and initial enzyme velocities, based on HFB1 deamination, were plotted.

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