Bifunctional Sesquiterpene/Diterpene Synthase Agr2 from Cyclocybe aegerita Gives Rise to the Novel Diterpene Cyclocybene

Summary

Scientists discovered that a special enzyme from a mushroom called Cyclocybe aegerita can make two different types of aromatic compounds instead of just one. By growing this enzyme in a different mushroom species that provides better raw materials, researchers identified a completely new compound called cyclocybene. This finding shows that using fungi as hosts for producing useful natural chemicals can work better than traditional bacterial systems, potentially opening new paths for making medicines and fragrances.

Background

Cyclocybe aegerita is a model mushroom known for its diverse terpenome containing various volatile and nonvolatile terpenes. The terpene synthase Agr2 was previously thought to produce only the sesquiterpene viridiflorene, but genomic evidence suggested it might also produce a diterpene.

Objective

To investigate the complete biosynthetic products of the terpene synthase Agr2 from C. aegerita and determine if it has bifunctional capacity to produce both sesquiterpenes and diterpenes.

Results

Heterologous expression revealed two terpenes: the known sesquiterpene viridiflorene and a novel diterpene identified as cyclocybene with molecular formula C20H32. Biotransformation experiments with substrates confirmed Agr2’s bifunctionality, with preferential conversion of GGPP to cyclocybene, achieving production levels of approximately 1.9 mg/L.

Conclusion

Agr2 is confirmed as a bifunctional terpene synthase capable of producing both sesquiterpenes and diterpenes, demonstrating that C. cinerea is a superior expression host compared to E. coli for fungal diterpene biosynthesis due to adequate GGPP availability.
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