Bivariate One Strain Many Compounds Designs Expand the Secondary Metabolite Production Space in Corallococcus...

Area of Science:

• Microbiology
• Natural Product Discovery
• Synthetic Biology

Background:

• The myxobacterium Corallococcus coralloides possesses a large genome with numerous uncharacterized biosynthetic gene clusters (BGCs) encoding potential novel natural products.
• Activating these BGCs for secondary metabolite (SM) production is challenging due to unknown optimal cultivation conditions, limiting the exploration of its biosynthetic potential.

Purpose of the Study:

• To identify effective cultivation strategies for activating silent BGCs in C. coralloides.
• To discover new secondary metabolites (SMs) by stimulating BGCs using a "one strain many compounds" (OSMAC) approach.

Main Methods:

• Applied univariate and bivariate "one strain many compounds" (OSMAC) screening in Duetz-System 24-well plates.
• Combined biotic additives and organic solvents with complex or minimal media under different elicitation conditions.
• Evaluated BGC activation by analyzing new mass features in bacterial extracts using mass spectrometry and molecular networking.

Main Results:

• Bivariate OSMAC designs, using multiple elicitors simultaneously, showed synergistic effects in activating BGCs.
• Detected 55 new mass features in bivariate OSMAC experiments, not observed in univariate conditions.
• Identified potential novel natural compounds, including N-acyl fatty amines and sulfur-containing products, through molecular network analysis.

Conclusions:

• Bivariate OSMAC designs with combined elicitors are a robust strategy to activate cryptic BGCs and expand the secondary metabolome of microorganisms.
• This approach successfully unlocked the biosynthetic potential of C. coralloides, paving the way for novel natural product discovery.