Discovering Dynamic Plant Enzyme Complexes in Yeast for Kratom Alkaloid Pathway Identification

Area of Science:

• Plant biochemistry
• Molecular biology
• Natural product discovery

Background:

• Identifying natural product biosynthetic pathways in medicinal plants is complex and time-consuming.
• Transcriptomic regulation and enzyme coregulation are established methods for pathway identification.
• Protein-protein interactions (PPI) offer a complementary approach to discovering pathway organization.

Purpose of the Study:

• To develop and validate a yeast-based screening method for capturing plant enzyme PPIs.
• To discover novel enzyme complexes and biosynthetic pathways in the alkaloid-producing plant, kratom.
• To integrate PPI data with multiomics analysis for enhanced pathway discovery.

Main Methods:

• Yeast-based protein-protein interaction (PPI) screening.
• Multiomics analysis of plant samples.
• Biochemical characterization of identified enzymes.

Main Results:

• Discovered four enzyme complexes and organized biosynthetic pathways in kratom.
• Identified six medium-chain dehydrogenase/reductases (MsMDRs) interacting with strictosidine β-D-glucosidase (MsSGD).
• Validated the function of four MsMDRs, demonstrating high PPI screening selectivity.

Conclusions:

• Leveraging protein-protein interactions is a powerful strategy for discovering plant natural product biosynthetic pathways.
• The developed yeast-based PPI screening method effectively complements multiomics approaches.
• This study opens new avenues for exploring post-translational regulation in pathway elucidation.