Figure 3.

Kinetic studies of EGFP-T1 were conducted in trypsin digestion buffer. The kinetic parameters, k_cat, K_m, and k_cat/K_m, were determined to be 0.26 ± 0.01/s, 3.99 ± 0.38 µM, and 65200 ± 5000 M⁻¹s⁻¹ through Michaelis-Menten equation fitting (a), respectively. EGFP-T1 with trypsin inhibitor, leupeptin with 0 nM (○), 12.5 nM (□), 25 nM (△), 50 nM (▽), 100 nM (⊠) is evaluated (b) to reveal the competitive inhibition to trypsin, with K_i value of 31 ± 2 nM.