Title
Variants of glycosyl hydrolase family 2 beta-glucuronidases
Conference Dates
September 15-19, 2019
Abstract
Mammals remove toxic metabolites and xenobiotics by attaching glucuronic acid to compounds targeted for excretion. The glucuronide increases the solubility of the compound, enabling secretion in urine or bile. Metabolites of opiates and opioids can be quantified from urine by hydrolyzing conjugates with β-glucuronidase (β-GUS) and separating free drug molecules by liquid chromatography and mass spectrometry (LC-MS). Importantly, the hydrolysis step increases both detection and quantitation because free drug molecules ionize better than conjugates in MS (Sitasuwan et al., 2019). However, not all glucuronidases have the same activity across a range of potential substrates. Therefore, we generated β-GUS variants with improved activity on a broader range of drug metabolites by using efficient domain swapping and site-saturation mutagenesis techniques.
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Recommended Citation
Caleb Schlachter, Pongkwan N. Sitasuwan, L. Andrew Lee, Amanda C. McGee, and John J. Tomashek, "Variants of glycosyl hydrolase family 2 beta-glucuronidases" in "Enzyme Engineering XXV", Huimin Zhao, University of Illinois at Urbana-Champaign, USA John Wong, Pfizer, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/enzyme_xxv/59