US 12,252,719 B2
Chimeric and other variant β-glucuronidase enzymes with enhanced properties
Caleb Reece Schlachter, Irmo, SC (US); John Tomashek, Columbia, SC (US); and Lim Andrew Lee, Columbia, SC (US)
Assigned to INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC., Irmo, SC (US)
Filed by INTEGRATED MICRO-CHROMATOGRAPHY SYSTEMS, INC., Irmo, SC (US)
Filed on Jul. 14, 2022, as Appl. No. 17/865,099.
Application 17/865,099 is a division of application No. 16/596,568, filed on Oct. 8, 2019, granted, now 11,421,210.
Claims priority of provisional application 62/742,779, filed on Oct. 8, 2018.
Prior Publication US 2023/0060620 A1, Mar. 2, 2023
Int. Cl. C12N 9/24 (2006.01); C12N 9/42 (2006.01); C12N 15/70 (2006.01); C12P 19/14 (2006.01)
CPC C12N 9/2402 (2013.01) [C07K 2319/00 (2013.01); C12N 9/2445 (2013.01); C12N 15/70 (2013.01); C12P 19/14 (2013.01); C12Y 302/01031 (2013.01)] 26 Claims
OG exemplary drawing
 
1. A variant Brachyspira pilosicoli beta-glucuronidase (BpBGUS) enzyme comprising an amino acid sequence at least 95% homologous to a parental BpBGUS amino acid sequence shown in SEQ ID NO: 5, wherein the variant comprises a substitution at amino acid position Q451, wherein the substitution is selected from the group consisting of aspartic acid (D), glutamic acid (E), serine(S), valine (V) and lysine (K), and wherein the variant exhibits an increased level of enzymatic activity for one or more substrates as compared to the parental BpBGUS enzyme, wherein the one or more substrates are selected from the group consisting of morphine-3-β-D-glucuronide (MOR), oxymorphone-3-β-D-glucuronide (OMOR), codeine-6-β-D-glucuronide (COD), dihydrocodeine-6-β-D-glucuronide (DCOD) and amitriptyline-N-β-D-glucuronide (AMT gluc).