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pH-dependent cyclization efficiency of VcAEP and mutants. (A–C) Quantitative summary of VcAEP mutant Vc1a-Vc1c product yield analyzed using MALDI-TOF mass spectrometry. Gray and black bars indicate the percentage yield of linear product and cyclic product, respectively. The reactions were conducted at 37°C for 15 min with a molar enzyme:substrate ratio of 1:200 (Vc1a), 1:500 (Vc1b), and 1:1000 (Vc1c). Average yield and SDs were calculated from experiments performed in triplicate.
