Functional Redundancy and Dual Function of a Hypothetical Protein in the Biosynthesis of Eunicellane-Type Diterpenoids

• DOI: 10.1021/acschembio.4c00413
• Publication Date: 2024-11-01T12:51:06Z
• Authors: Ayesha Ahmed Chau...; Yuya Kakumu; Sirinthra Thiengmag; Jack Chun-Ting Liu; Geng-Min Lin; Suhan Durusu; Friederike Biermann; Miriam Boeck; Christopher A. Voigt; Jon Clardy; Reiko Ueoka; Allison S. Walker; Eric J. N. Helfrich
• Keywords: Dual function

Many complex terpenoids, predominantly isolated from plants and fungi, show drug-like physicochemical properties. Recent advances in genome mining revealed actinobacteria as an almost untouched treasure trove of terpene biosynthetic gene clusters (BGCs). In this study, we characterized a BGC with unusual architecture. The selected includes, among others, genes encoding cyclase fused to truncated reductase domain cytochrome P450 monooxygenase (P450) that is split over three fragments. Functional characterization the heterologous host led identification several new members trans-eunicellane family diterpenoids, euthailols, feature unique oxidation patterns. A combination bioinformatic analyses, structural modeling studies, expression dual function pathway-encoded hypothetical protein acts isomerase oxygenase. Moreover, absence other tailoring enzymes, hydroxylates eunicellane scaffold at position not modified eunicellanes. Surprisingly, both modifications installed by one P450s exhibit partial redundancy. Bioactivity assays some euthailols growth inhibitory properties against Gram-negative ESKAPE pathogens. euthailol study provides unprecedented insights into functional redundancy enzymes diterpenoid biosynthesis highlights proteins important largely overlooked involved maturation terpenoids.