Method of synthesis of testosteronan polymer and derivatives and uses thereof

Recombinant microbial platforms using engineered Testan synthases in heterologous hosts achieve high-molecular-weight, low-polydispersity Testan production, addressing limitations in existing methods and enabling scalable production of functional derivatives for chromatography and biomaterials.

WO2026112535A1PCT designated stage Publication Date: 2026-05-28WEKA BIOSCIENCES LLC
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Patent Information

Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
WEKA BIOSCIENCES LLC
Filing Date
2025-11-21
Publication Date
2026-05-28

AI Technical Summary

Technical Problem

Existing methods for producing Testosteronan (Testan) polysaccharides are limited in achieving high molecular weight, low polydispersity, and functionalized derivatives suitable for chromatography and biomaterials, lacking scalable and recombinant production systems with confirmed structural identity and rheological properties.

Method used

Development of recombinant microbial platforms using engineered Testan synthases, such as CtTS and CIPTS, in heterologous hosts like E. coli and Bacillus, with optimized codon sequences and promoters, enabling high-molecular-weight, low-polydispersity Testan production and functional derivatives like sulfated Testan, through fermentation and purification processes.

Benefits of technology

The recombinant systems produce chemically identical Testan with molecular weights exceeding 800 kDa and PDI approaching 1.02, suitable for chromatography and biomaterials, providing scalable and sustainable solutions for biomedical and industrial applications.

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Abstract

Embodiments of the present invention provides recombinant systems, methods, and compositions for the biological production of Testosteronan (Testan), a linear α-(1→4)-linked glucuronic acid–N-acetylglucosamine polysaccharide. Microbial hosts engineered to express full-length CtTS, N-terminal truncation variants (including d64-CtTS), or the Pseudomonas CIPTS homolog synthesize high–molecular-weight Testan (≥800 kDa) with low polydispersity. Structural analysis by ¹H NMR and 2D HSQC demonstrates that all recombinant enzymes produce Testan that is chemically indistinguishable from native polymer. Purification using ultrafiltration and strong-anion exchange chromatography yields highly defined molecular-weight fractions, including monodisperse populations (PDI ~1.02). The invention further provides sulfated derivatives of Testan produced via aqueous sulfation methods, as well as Testan and functionalized Testan compositions useful for chromatographic stationary phases, biomaterial scaffolds, and chemically modifiable polymer platforms. These systems provide the first scalable, fermentation-based production methods for Testan and Testan-derived materials.
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