Production of pseudaminic acids and their CMP-activated derivatives

The National Institute of General Medical Sciences awarded Midwest Bioprocessing Center, Inc. $313,770 on September 1, 2026, under the SBIR program to develop a scalable fermentation system for producing pseudaminic acids and their CMP-activated derivatives.

The award supports Phase I research to establish proof-of-concept for cost-effective biosynthetic production of pseudaminic acid compounds—specifically PSE5AC7AC and analogs PSE5AC7HB and PSE5AC7FM in free and CMP-activated forms. Pseudaminic acids are non-mammalian sugars found in surface glycoconjugates of major pathogens including Pseudomonas aeruginosa, Helicobacter pylori, Campylobacter jejuni, and Acinetobacter baumannii. They function as critical virulence factors and are targets for antimicrobial drug discovery, glycoconjugate vaccine development, and pathogen detection. Current commercial availability is severely limited; PSE5AC7AC and CMP-PSE5AC7AC cost more than $4,000 to $6,000 per milligram, and other derivatives are not commercially available.

The project will engineer Escherichia coli fermentation strains expressing five biosynthetic genes from Aeromonas caviae to produce and secrete CMP-activated pseudaminic acids directly into culture media. Phase I milestones include cloning and expressing the biosynthetic genes, co-expressing them in a single E. coli strain to achieve gram-scale production of CMP-PSE5AC7AC, developing purification protocols, and isolating both CMP-PSE5AC7AC and PSE5AC7AC to demonstrate proof-of-concept for research-grade production scales.

Work is performed in Peoria, Illinois. The period of performance runs from September 1, 2026, through February 28, 2027.

Abstract

The goal of this phase I sbir project is to develop a scalable system to produce pseudaminic acid (PSE) compounds-specifically PSE5AC7AC and its analogs PSE5AC7HB and PSE5AC7FM-in both free and CMP- activated forms. Pseudaminic acids are non-mammalian sugars found in surface glycoconjugates of major pathogens such as pseudomonas aeruginosa, helicobacter pylori, campylobacter jejuni, and acinetobacter baumannii. They are essential components of bacterial structural glycans and glycoproteins, play critical roles in flagellar assembly and motility, and act as key virulence factors. Their unique biology makes them attractive targets for antimicrobial drug discovery, glycoconjugate vaccine development, and pathogen detection strategies. Despite their importance, access to pseudaminic acid compounds remains extremely limited due to the complexity of existing chemical syntheses and the high cost of commercial sources (>$4,000-6,000 per MG for PSE5AC7AC and CMP-PSE5AC7AC). Other derivatives are not commercially available at all, creating a significant barrier for biomedical research and translational applications. Our proposed solution is to engineer escherichia coli fermentation strains expressing the five biosynthetic genes from aeromonas caviae to produce and secrete CMP-activated pseudaminic acids directly into culture media, from which they can be purified and, if needed, hydrolyzed to yield the free sugar. During phase I, we will establish feasibility by (1) cloning and expressing the biosynthetic genes, (2) CO- expressing them in a single E. Coli strain to produce CMP-PSE5AC7AC at gram scale, (3) developing a purification protocol, and (4) isolating both CMP-PSE5AC7AC and PSE5AC7AC. These milestones will demonstrate proof-of-concept for cost-effective production at scales suitable for research applications. In phase II, we will optimize yields and expand production to additional analogs (CMP-PSE5AC7HB and CMP- PSE5AC7FM), and in phase III we will commercialize these compounds as research reagents and partner in translational development of novel antimicrobial and vaccine strategies.