Engineering Baker’s Yeast for Efficient cAMP Synthesis via Regulation of PKA Activity

TL;DR

Scientists engineered baker's yeast to produce more cAMP, a molecule important for cell function, by modifying genes that control its activity.

Contribution

A novel strategy for boosting cAMP biosynthesis in yeast through gene regulation is demonstrated.

Findings

Engineered yeast strains BN5-126 and BN5-310 showed 30- and 9-fold higher cAMP content compared to the wild strain.

TPK1 gene mRNA levels were reduced in engineered strains without affecting yeast growth.

Modified strains exhibited enhanced PKA activity and improved heat shock tolerance.

Abstract

cAMP (cyclic adenosine-3′,5′-monophosphate) has extensive physiological functions and nutritional value for living organisms, and it regulates cellular metabolism mainly by modulating PKA (protein kinase A) activity. The current yields of cAMP synthesized by microbial fermentation are still low, which is arousing interest in developing high-yield cAMP strains. In this work, two baker’s yeasts with high cAMP content were constructed by knocking out BCY1, TPK3, and TPK2 genes, and truncating the promoter of the TPK1 gene. The content of cAMP in BN5-126 and BN5-310 (with the TPK1 gene promoter truncated by 126 and 310 bp in BN5) was improved by 30- and 9-fold, respectively, relative to the wild strain. The TPK1 gene mRNA levels of BN5-126 and BN5-310 were decreased by 18% and 40%, respectively, without significant changes in growth performance. The results of heat shock tolerance of engineered strains also reflected the enhanced PKA activity. This work demonstrates a novel strategy for regulating gene expression to boost cAMP biosynthesis in yeast, providing a promising platform for producing nutritionally enriched and functional fermented products.

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