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The 9-GHz EPR spectra of Trp• in the (E250Q+E168Q) variant of LiP (red trace) obtained by the reaction with 5-fold excess H2O2 (2-s mixing time at 0 °C). The spectrum of the enzyme premixed with 2-fold excess of VA before the reaction with H2O2 showed no Trp• signal. Only a very narrow radical (peak-to-trough of 12 G) was detected (black trace; Inset); the same species was already detected in the reaction of the WT LiP with H2O2 (gray trace; Inset).
