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Bioresources and Bioprocessing
Fig. 2 | Bioresources and Bioprocessing

Fig. 2

From: Reduction of phenolics in faba bean meal using recombinantly produced and purified Bacillus ligniniphilus catechol 2,3-dioxygenase

Fig. 2

Purification and characterization of recombinant BLC23O. A Ni–NTA Affinity purification of recombinant BLC23O. SDS–PAGE showing the obtained pellet (P) and supernatant (S) fractions from an E. coli lysate. Subsequent flow-through (FT), washes (W) and elutions (E) are included. The elution fractions were pooled and used in all subsequent experiments. A small portion of the pooled elution was further purified by SEC (Additional file 1: Figure S1). B Kinetic analysis of Ni–NTA + SEC purified BLC23O. Following SEC purification, a Michaelis–Menten kinetic analysis of BLC23O catalytic activity was conducted. BLC23O (18.7 µg/mL) was combined with 10 mM of the co-factor Mn2+ and varying concentrations of the substrate 3-methylcatechol. The data were fitted to the Michaelis–Menten equation, using Microsoft Excel’s Solver

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