Analyzing and Engineering the Product Selectivity of a 2-Methylenebornane Synthase

Area of Science:

• Natural Product Chemistry
• Biotechnology
• Enzymology

Background:

• Terpenes are a vast class of natural products with diverse applications.
• Bacterial pathways can produce noncanonical C11 terpenes, but often in minor yields.
• Exploring these less common terpenes is crucial for new chemical and pharmaceutical discoveries.

Purpose of the Study:

• To alter the product selectivity of 2-methylenebornane synthase from Pseudomonas fluorescens.
• To enable the biotechnological synthesis of unexplored C11 terpenes.
• To gain insights into the structure-function relationships of C11 terpene synthases.

Main Methods:

• Semirational protein engineering of terpene synthase active sites.
• Identification and characterization of enzyme variants.
• Analysis of altered product spectra.

Main Results:

• Successfully modified the product selectivity of the target terpene synthase.
• Identified key amino acids influencing product specificity.
• Generated variants with significantly altered and novel terpene product profiles.

Conclusions:

• Protein engineering can redirect terpene synthase pathways to produce specific noncanonical C11 terpenes.
• This work provides a foundation for accessing a wider range of C11 terpenes.
• Demonstrated the potential for producing previously inaccessible natural products through enzyme modification.