ZFIN Image: Küssau et al., 2020, Figure 3

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Figure Caption

Figure 3

Biochemical characterization of Ami1_Mab. (A) Ami1_Mab elution profile on Superdex 75 10/300 GL size-exclusion chromatography column. The denaturating polyacrylamide electrophoresis gel attests to the high protein purity after three chromatography steps. (B) Schematic of the muramyl-dipeptide hydrolysis assay. Ami1_Mab cleaves muramyl-dipeptide (MDP) to yield N-acetylmuramic acid (MurNAc) and l-Ala-d-IsoGln (dipeptide). (C) The thin-layer chromatography (TLC)-based activity assay revealed a correlation between the disappearance of MDP and the concentration of Ami1_Mab. Pure MDP, MurNAc, and dipeptide (DP) were loaded on the right part of the TLC plate as migration controls. TLC was revealed by ninhydrin and charring. (D) The enzymatic activity of Ami1 is Zn²⁺-dependent. Activity of both Ami1_Mab and Ami1_Mtb is inhibited after incubation with EDTA.

Acknowledgments

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