New Prodigiosin Derivatives Obtained by Mutasynthesis in Pseudomonas putida

Area of Science:

• Natural product chemistry
• Microbial biosynthesis
• Synthetic biology

Background:

• Prodigiosin and related prodiginines are alkaloids with antimicrobial, antitumor, and antimalarial properties.
• Bacterial production of prodiginines, particularly by Serratia marcescens, often yields complex mixtures, complicating purification and cost-effectiveness.
• Structural variations in prodiginines correlate with differential biological activities.

Purpose of the Study:

• To develop a mutasynthesis approach for generating diverse prodiginine derivatives.
• To investigate the substrate specificity of the PigC enzyme in prodigiosin biosynthesis.
• To evaluate the biological activity, specifically autophagy modulation, of novel prodiginine derivatives.

Main Methods:

• Utilized a mutasynthesis strategy in the GRAS-classified strain Pseudomonas putida KT2440.
• Employed genetic engineering of the prodigiosin biosynthetic pathway.
• Incorporated synthetic intermediates to create new prodiginine structures.

Main Results:

• Successfully produced 13 distinct prodigiosin derivatives in P. putida, demonstrating unprecedented structural diversity.
• Gained insights into the substrate specificity of PigC, the final enzyme in prodigiosin biosynthesis.
• Confirmed that the autophagy-modulating activity of prodigiosin is retained in its derivatives.
• Identified one derivative with superior autophagy inhibitory activity compared to prodigiosin and obatoclax.

Conclusions:

• The mutasynthesis approach in P. putida is effective for generating diverse prodiginine analogs.
• The study provides valuable information on PigC enzyme function.
• Prodiginine derivatives retain significant biological activity, with potential for enhanced therapeutic efficacy in autophagy-related conditions.