MYCOLab Ai Logo - gold and army green

Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae

Summary

Researchers developed new tools to make it faster and easier to discover useful compounds from fungi. They created improved genetic vectors that allow scientists to insert multiple genes into Aspergillus oryzae more conveniently, and developed a quick screening method using mass spectrometry to identify successful transformants directly on culture plates. This approach saves about 10 days compared to traditional methods, significantly accelerating the discovery of new natural products with potential medical and agricultural applications.

Background

Aspergillus oryzae is widely used as a host for heterologous expression of fungal natural products. However, previously developed vectors lack convenience and screening positive transformants through PCR and fermentation is time-consuming and labor-intensive.

Objective

To develop plug-and-play vectors for multi-gene expression in Aspergillus oryzae and introduce LC–MS detection as a more efficient method for screening positive transformants, saving significant time in the natural product discovery process.

Results

The developed vectors successfully expressed the rug BGC producing rugulosin A, with LC–MS detection identifying positive transformants directly from agar plates. The P xyrA promoter proved superior to P amyB for regulating deleterious gene expression. The toolkit saved approximately 10 days in the discovery process compared to traditional methods.

Conclusion

The plug-and-play vectors and LC–MS screening strategy significantly accelerate natural product discovery in Aspergillus oryzae by eliminating the need for intermediate cloning steps and enabling direct detection on solid medium without fermentation, while P xyrA provides tight regulation for deleterious genes.
  • Published in:Bioresource and Bioprocessing,
  • Study Type:Research Article,
  • Source: 10.1186/s40643-024-00833-w, PMID: 39776300
Scroll to Top