Understanding mechanisms of terpene synthases using substrate analogs

Area of Science:

• Biochemistry
• Organic Chemistry
• Enzymology

Background:

• Terpene synthases (TS) catalyze complex hydrocarbon scaffold formation from prenyl substrates.
• These reactions involve high-energy carbocation intermediates crucial for product diversity.
• Understanding TS mechanisms is vital for synthetic biology and drug discovery.

Purpose of the Study:

• To investigate terpene synthase (TS) mechanisms using novel fluorinated substrate analogs.
• To explore the synthesis and application of fluorine-containing analogs for mechanistic studies.
• To elucidate the role of carbocation intermediates in TS-catalyzed reactions.

Main Methods:

• Synthesis of specific substrate analogs with strategic fluorine atom replacement.
• Utilizing these analogs to probe carbocation intermediate generation and stabilization by TS.
• Analyzing reaction pathways and product formation influenced by fluorination.

Main Results:

• Fluorinated analogs effectively inhibit the formation of specific carbocation intermediates.
• The study provides insights into the stabilization mechanisms of transient carbocations within the TS active site.
• Demonstration of fluorine as a tool to dissect complex enzymatic reaction pathways.

Conclusions:

• Fluorinated substrate analogs are powerful tools for studying terpene synthase mechanisms.
• Understanding carbocation stabilization is key to controlling TS product outcomes.
• This approach offers new avenues for enzyme engineering and the development of novel terpenes.