Kinetic activity of recombinant KpACE.

<p><b>(A)</b> Size-exclusion chromatography purification of recombinant KpACE. Purification of the rKpACE protein using molecular size Superdex 200. Samples corresponding to the peak position at 14.38 mL (75.6 kDa) were collected, and the presence of the KpACE protein was confirmed by 12% SDS-PAGE gel electrophoresis followed by protein staining. <b>(B)</b> Kinetic acetylesterase activity of rKpACE determined with 4-MU-Ac at pH 7.4. The data were nonlinearly fitted to the Michaelis-Menten equation by GraphPad Prism v9.3.1. <b>(C)</b> Hydrolytic activities of KpACE on three fully acetylated monosaccharides. Acetylated monosaccharides include 1,2,3,4-Tetra-O-acetyl-β-D-xylopyranose (Xyl), 1,2,3,4,6-Penta-O-acetyl-β-D-galactopyranose (Gal), 1,2,3,4,6-β-D-glucose pentaacetate (Glu). The activity was determined by measuring the amounts of acetic acids released from the substrates by rKpACE and its variants. Data are from six biological replicates. The unpaired <i>t-test</i> was performed to determine significant differences.</p> <p>(PDF)</p>