Fine-tuned synthetic transcription factors for production of 3'-phosphoadenosine-5'-phosphosulfate in yeast.

Authors

Madhushruti Borah, Shanna Gu, Essa M Saied, Christoph Arenz, Mattheos Koffas, Gita Naseri

Published in

Microbial cell factories. Volume 25. Issue 1. Jul 27, 2026. Epub Jul 27, 2026.

Abstract

Technologies developed over the past decade have made Saccharomyces cerevisiae a promising platform for producing various natural products. Balancing multi-enzyme expression, while maintaining robust microbial growth, remains a limiting factor for engineering long biosynthetic pathways in yeast.
Here, we improved the transcriptional capacity of our previously developed synthetic transcription factors (synTFs) derived from the plant JUB1 DNA-binding domain, whose expression is controlled by an β-D-1-thiogalactopyranoside (IPTG)-inducible promoter. To this end, at cysteine positions within surface-exposed loop regions of a JUB1-derived DNA-binding scaffold, we introduced a short peptide to enhance loop flexibility while providing local stability and orientation. The generated synTFs, so-called JUB1-X synTFs, varying in strength, have been successfully used to improve the production of 3'-phosphoadenosine 5'-phosphosulfate (PAPS), a universal sulfate donor necessary for the synthesis of therapeutic glycosaminoglycans and sulfolipids, in yeast. Using the generated yeast strain, in simple batch culture, PAPS accumulation of 21.4 ± 5.8 mg g⁻¹ cdw was achieved after only 5 h of inducing the expression of JUB1-X synTFs.
The design principle demonstrated here provides a generalizable strategy to fine-tune other synTFs derived from heterologous DNA-binding domains, expanding the regulatory capabilities of existing synTF collections. Together, this work offers a modular, scalable approach to constructing high-performance gene circuits and supports the development of yeast cell factories for the synthesis of complex natural products.

PMID:
42509562
Bibliographic data and abstract were imported from PubMed on 28 Jul 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.