Efficient glycyrrhetinic acid biomanufacturing through protein engineering and dual-GUS combination strategy with novel β-glucuronidase from Aspergillus calidoustus CLH-22

Glycyrrhetinicacid (GA) is a high-value pentacyclic triterpenoid with broad applications. However, the industrial production of GA is hindered by low yield and the accumulation of the intermediate product GlycyrrhetinicAcid3-O-Mono-β-D-Glucuronide (GAMG). This study first identified a novel β-glucuronidase (AcGUS) from Aspergillus calidoustus CLH-22 through transcriptomic analysis, demonstrating a substrate preference for GAMG. Subsequently, mutant AcGUS3