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Fermentative aminopyrrolnitrin production by metabolically engineered Corynebacterium glutamicum
Virginia Ryandini Melati Putri1, Min-Hee Jung1, Ji-Young Lee1
1Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea.
Microbial Cell Factories
|May 23, 2024
Summary
This study engineered Corynebacterium glutamicum for microbial production of aminopyrrolnitrin (APRN), a potent antifungal and antiparasitic compound. The engineered strain achieved significant yields of APRN, paving the way for its scalable fermentation.
Area of Science:
- Microbiology
- Biotechnology
- Synthetic Biology
Background:
- Aminopyrrolnitrin (APRN) is a halogenated phenylpyrrole derivative with potent antifungal and antiparasitic activities.
- APRN exhibits enhanced photostability compared to existing antimicrobial compounds like pyrrolnitrin.
- Developing efficient microbial production methods for APRN is crucial for its wider application.
Purpose of the Study:
- To establish a metabolically engineered Corynebacterium glutamicum strain for the fermentative production of APRN.
- To optimize the biosynthetic pathway for APRN by combining and expressing key enzymes from different microbial sources.
- To achieve significant yields of APRN through a multi-step metabolic engineering approach.
Main Methods:
- Engineering of anthranilate phosphoribosyltransferase (TrpD) for feedback-resistant L-tryptophan production.
- Introduction and expression of genes encoding tryptophan 7-halogenase (PrnA/RebH) and flavin reductase (Fre/PrnF/RebF) for 7-chloro-L-tryptophan synthesis.
- Expression of monodechloroaminopyrrolnitrin (MDAP) synthase (PrnB) and MDAP halogenase (PrnC) to complete the APRN biosynthetic pathway.
- Fermentative production of APRN in engineered C. glutamicum strains.
Main Results:
- Engineered C. glutamicum produced 3.1 g/L of L-tryptophan using a feedback-resistant TrpD variant.
- Optimized combinations of halogenase and reductase genes led to efficient 7-chloro-L-tryptophan production.
- The final engineered strain successfully produced 29.5 mg/L of APRN via fermentation.
- This marks the first report of fermentative APRN production using metabolically engineered C. glutamicum.
Conclusions:
- Metabolic engineering of C. glutamicum provides a viable platform for the microbial production of APRN.
- The constructed artificial biosynthetic pathway enables efficient conversion of L-tryptophan to APRN.
- This study lays the foundation for scalable and sustainable production of APRN for antifungal and antiparasitic applications.
