Genome mining and characterization of bifunctional Clerodane Diterpene synthase from a fungus Myrothecium sp

Shuxuan Li; Ke Ma; Yujie Zhao; Luning Zhou; Peng Zhang; Hanwei Liu; Yajin Ye; Wenhan Lin; Jaclyn M Winter; Guangwei Wu

doi:10.1016/j.bioorg.2025.108548

Genome mining and characterization of bifunctional Clerodane Diterpene synthase from a fungus Myrothecium sp

Bioorg Chem. 2025 Jul 1:161:108548. doi: 10.1016/j.bioorg.2025.108548. Epub 2025 Apr 29.

Authors

Shuxuan Li¹, Ke Ma², Yujie Zhao³, Luning Zhou⁴, Peng Zhang⁵, Hanwei Liu⁶, Yajin Ye³, Wenhan Lin², Jaclyn M Winter⁷, Guangwei Wu⁸

Affiliations

¹ Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China.
² Ningbo Institute of Marine Medicine, Peking University, Ningbo 315832, China.
³ State Key Laboratory of Tree Genetics and Breeding, Co-Innovation Center for Sustainable Forestry in Southern China, Jiangsu Key Laboratory for Poplar Germplasm Enhancement and Variety Improvement, Nanjing Forestry University, Nanjing 210037, China.
⁴ Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
⁵ School of Life Sciences, Inner Mongolia University, Hohhot, 010010, China; Department of Pharmacology and Toxicology, College of Pharmacy, University of Utah, Salt Lake City, UT 84112, USA.
⁶ Ningbo Customs District Technology Center, Ningbo 315100, China.
⁷ Department of Pharmacology and Toxicology, College of Pharmacy, University of Utah, Salt Lake City, UT 84112, USA. Electronic address: jaclyn.winter@utah.edu.
⁸ Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China; Ningbo Institute of Marine Medicine, Peking University, Ningbo 315832, China. Electronic address: gweiwu@njfu.edu.cn.

PMID: 40315625
DOI: 10.1016/j.bioorg.2025.108548

Abstract

Clerodane diterpenoids are a promising group of pharmacological molecules and are rarely discovered in fungi. The clerodane diterpene synthase has not yet been characterized in fungi. Herein, we discovered a gene, mterA, encoding chimeric protein with the bifunction of class II and class I synthases from a fungus Myrothecium sp. by genome mining. Heterologous co-expression of mterA and mterB in engineered yeast led to the production of a clerodane diterpene product (5S, 8R, 9R, 10S)-terpentetriene (1), confirming the existence of a fungi-derived special clerodane diterpene synthase. Site-directed mutagenesis on conserved motifs revealed a strong influence on terpentetriene accumulation. Gene inactivation of mterA indicated that the mter gene cluster was likely cryptic or weakly expressed in the host under the laboratory conditions. Compound 1 exhibited significant inhibitory activity against the plant pathogen Bacillus subtilis with a MIC value of 8 μg/mL and activated the plant immune response. MterA representes the first chimeric bifunctional clerodane diterpene synthase.

Keywords: (5S, 8R, 9R, 10S)-terpentetriene; Clerodane; Diterpene synthases; Heterologous expression; Site-directed mutagenesis.

MeSH terms

Alkyl and Aryl Transferases* / genetics
Alkyl and Aryl Transferases* / metabolism
Bacillus subtilis / drug effects
Diterpenes, Clerodane* / chemistry
Diterpenes, Clerodane* / metabolism
Diterpenes, Clerodane* / pharmacology
Genome, Fungal
Microbial Sensitivity Tests
Molecular Structure
Structure-Activity Relationship

Substances

Diterpenes, Clerodane
Alkyl and Aryl Transferases