Identification, Heterologous Production and Bioactivity of Lentinulin A and Dendrothelin A, Two Natural Variants of Backbone N-methylated Peptide Macrocycle Omphalotin A

Summary

This research discovered and produced two new peptide molecules from mushrooms that could help protect plants from harmful parasitic worms. The scientists developed a way to make these complex molecules in yeast cells, which is more efficient than extracting them from mushrooms. This work has important implications for agriculture and drug development. Impacts on everyday life: – Could lead to new environmentally-friendly pesticides to protect crops from nematode parasites – Demonstrates a sustainable way to produce complex molecules using yeast instead of chemical synthesis – Opens possibilities for developing new peptide-based medicines with improved properties – Helps understand how mushrooms naturally defend themselves against pests – Could reduce reliance on harmful chemical pesticides in agriculture

Background

Fungi produce diverse bioactive natural products used as therapeutics for infectious diseases, cancer, and as immunomodulatory agents. Recent discoveries have highlighted novel fungal peptide natural products including non-ribosomal peptides (NRPs) and ribosomally synthesized and post-translationally modified peptides (RiPPs). Backbone N-methylation and macrocyclization improve peptide pharmacological properties by enhancing proteolytic stability, membrane permeability and target selectivity.

Objective

To identify, produce and characterize two novel peptide natural products (lentinulin A and dendrothelin A) that are homologous to the known nematotoxic peptide omphalotin A, using a heterologous expression system in Pichia pastoris yeast.

Results

The study successfully produced omphalotin A and two novel peptides – lentinulin A and dendrothelin A – in P. pastoris. All three peptides were ninefold backbone N-methylated macrocycles with similar sequences. Both omphalotin A and lentinulin A showed strong nematotoxicity against M. incognita with LC50 values of 0.840 μM and 0.341 μM respectively after 1 day, while dendrothelin A showed reduced activity. The peptides were also detected in extracts from the original fungal species, confirming their natural production.

Conclusion

The study established a synthetic biology platform for producing backbone N-methylated peptide macrocycles in P. pastoris. This system enabled the discovery and characterization of two novel peptide natural products with potential applications as nematicides. The platform can be used to produce and study other borosin peptides and generate new peptide variants with improved pharmacological properties.
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