Regulator-Guided Strain Prioritization and Genome Engineering Uncovers Concanamycin Analogs

Area of Science:

• Natural Product Chemistry
• Microbial Biotechnology
• Chemical Biology

Background:

• Bafilomycin and concanamycin analogs are plecomacrolide natural products with potent vacuolar-type H+-ATPase (V-ATPase) inhibitory activity.
• Limited accessibility to these scaffolds hinders comprehensive bioactivity evaluation and therapeutic development.
• Exploring microbial biosynthetic pathways is crucial for discovering novel analogs.

Purpose of the Study:

• To expand the chemical diversity of natural concanamycin analogs through an integrative approach.
• To identify and characterize novel concanamycin-like metabolites from *Streptomyces humidus*.
• To develop scalable strategies for natural product discovery and production.

Main Methods:

• Genome mining to identify a unique concanamycin-like biosynthetic gene cluster in *Streptomyces humidus*.
• Strain engineering coupled with untargeted metabolomics and molecular networking for metabolite dereplication and prioritization.
• Optimization of cultivation conditions for enhanced production of target compounds.

Main Results:

• Isolation of five known concanamycin analogs and three new analogs: tanamycin E, X, and F.
• Identification of a novel concanamycin-like biosynthetic gene cluster with unique genetic architecture.
• High-yield production of known (93.0 mg/L) and new (24.7 mg/L) compounds achieved through optimized cultivation.

Conclusions:

• An integrative strategy combining genome mining, strain engineering, and molecular networking effectively expands natural product chemical space.
• This approach overcomes traditional limitations in the production and discovery of bioactive natural products.
• The identified tanamycin analogs represent promising new leads for V-ATPase inhibitor research.