Alanine supplementation enhancing cordycepin production in Cordyceps militaris via upregulation of Cns2 and Cns3 genes expression levels.

Cordycepin, a key bioactive compound produced by Cordyceps militaris, faces the challenge of low productivity for commercial use. In this study, alanine supplementation in Cordyceps militaris boosted cordycepin production, peaking at 3 mg/g with 12 g/L concentration. Transcriptome analysis revealed 1711 differentially expressed genes, Pathway analysis indicates that protein processing in the endoplasmic reticulum was the most affected pathway. In addition, the transcriptome showed that adenylosuccinate lyase is essential for the synthesis of cordycepin. The modulation of four genes (Cns1-4) points to a regulatory mechanism that could increase cordycepin biosynthesis, offering a strategy to overcome low productivity for commercial applications.