Ergopeptines or their derivatives are widely used for treating neurodegenerative and cerebrovascular diseases.The nonribosomal peptide synthetasedD-lysergyl peptide synthetase A(LPSA)determines ergopeptine formation b...Ergopeptines or their derivatives are widely used for treating neurodegenerative and cerebrovascular diseases.The nonribosomal peptide synthetasedD-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,a-ergocryptine,and b-ergocryptine,precisely controlled by the substrate specificity of its three modules.Cp-LPSA2 was initially inactive but could be restored to catalyze a-ergosine formation.Using this platform,we validated that P1-LPSA1 and P1-LPSA2 from the reported C.purpurea P1 strain catalyzed ergotamine and a-ergocryptine formation,respectively.Typically,the nonribosomal 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 a-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.展开更多
基金supported by the Beijing Natural Science Foundation(Grant no.7252205)the CAMS Innovation Fund for Medical Sciences(Grant no.CIFMS2021-I2M-1-029)the National Natural Science Foundation of China(Grant no.81603002).
文摘Ergopeptines or their derivatives are widely used for treating neurodegenerative and cerebrovascular diseases.The nonribosomal peptide synthetasedD-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,a-ergocryptine,and b-ergocryptine,precisely controlled by the substrate specificity of its three modules.Cp-LPSA2 was initially inactive but could be restored to catalyze a-ergosine formation.Using this platform,we validated that P1-LPSA1 and P1-LPSA2 from the reported C.purpurea P1 strain catalyzed ergotamine and a-ergocryptine formation,respectively.Typically,the nonribosomal 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 a-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.
