Kinetic Resolution of 3-Fluoroalanine Using a Fusion Protein of D-Amino Acid Oxidase with Vitroscilla Hemoglobin

Abstract

In this study, a fusion protein (VHb-DAAO) of D-amino acid oxidase (DAAO) with Vitreoscilla hemoglobin (VHb) was functionally expressed in Escherichia coli and purified. The k_cat value VHb-DAAO (47.1 s⁻¹) towards rac-3-flouroalanine was about 2-fold higher than that of DAAO (21.9 s⁻¹). rac-3-Flouroalanine (500 mM) was kinetically resolved into (R)-3-fluoroalanine with high enatiomeric excess (>99%) by VHb-DAAO with about 52% conversion.