Aminoacyl-tRNA synthetases remove (proofread) incorrect substrates and thereby prevent errors in protein synthesis. We report enzyme-catalyzed pretransfer editing by pimeloyl-coenzyme A (CoA) ligase (BioW), a biotin synthetic enzyme that converts pimelate, a seven-carbon dicarboxylic acid, to its CoA ester. The noncognate BioW substrate glutaric acid results in hydrolysis of ATP to AMP with formation of only trace amounts of glutaryl-CoA, thereby mimicking pretransfer editing of incorrect aminoacyl-adenylates by aminoacyl-tRNA synthetases.