Unveiling nonribosomal peptide synthetases from the ergot fungus Claviceps purpurea involved in the formation of diverse ergopeptines.
10.1016/j.apsb.2025.03.022
- Author:
Jing-Jing CHEN
1
;
Ting GONG
1
;
Wei-Bo WANG
1
;
Tian-Jiao CHEN
1
;
Jin-Ling YANG
1
;
Ping ZHU
1
Author Information
1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
- Publication Type:Journal Article
- Keywords:
Biosynthetic mechanism;
Chimeric enzymes;
Directed biosynthesis;
Ergopeptine;
Heterologous expression;
NRP codes;
Nonribosomal peptide (NRP) synthetase;
d-Lysergyl peptide synthetase A
- From:
Acta Pharmaceutica Sinica B
2025;15(6):3321-3337
- CountryChina
- Language:English
-
Abstract:
Ergopeptines or their derivatives are widely used for treating neurodegenerative and cerebrovascular diseases. The nonribosomal peptide synthetase-d-lysergyl peptide synthetase A (LPSA) determines ergopeptine formation but the detailed mechanism remains to be elucidated. Here, we characterized two LPSAs from Claviceps purpurea Cp-1 strain through heterologous expression in Aspergillus nidulans feeding with d-lysergic acid. We proved that Cp-LPSA1 catalyzed the formation of ergocornine, α-ergocryptine, and β-ergocryptine, precisely controlled by the substrate specificity of its three modules. Cp-LPSA2 was initially inactive but could be restored to catalyze α-ergosine formation. Using this platform, we validated that P1-LPSA1 and P1-LPSA2 from the reported C. purpurea P1 strain catalyzed ergotamine and α-ergocryptine formation, respectively. Typically, the non-ribosomal peptide codes implicated in every module of the LPSAs were defined and elucidated, in which certain key residues could play a switched role for substrate specificity and product interconversion. By constructing chimeric LPSAs through module assembly, the production of the desired ergopeptines was achieved. Notably, 1.46 mg/L of α-ergocryptine and 1.09 mg/L of ergotamine were produced respectively by mixed-culture of C. paspali No. 24 (fermentation supernatant) and the recombinants of A. nidulans. Our findings provide insights into the biosynthetic mechanism of ergopeptines and lay a foundation for directed ergopeptine biosynthesis.
