Related Experiment Video
Updated: May 31, 2026
07:59
A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
Published on: October 4, 2019
Computational design and selections for an engineered, thermostable terpene synthase.
Juan E Diaz1, Chun-Shi Lin, Kazuyoshi Kunishiro
1Department of Chemistry, University of California, Irvine, California 92697-2025, USA.
Summary
Scientists engineered a thermostable terpene synthase (TEAS) mutant for high-temperature catalysis. This enhanced enzyme retains activity at 65°C, expanding possibilities for synthesizing valuable terpenoids.
Area of Science:
- Biochemistry
- Enzyme Engineering
- Synthetic Biology
Background:
- Terpenoids are diverse natural products with applications in medicine and industry.
- Engineering terpene synthases (TEAS) is crucial for controlling the synthesis of specific terpenoids.
- Current limitations include enzyme stability at elevated temperatures.
Purpose of the Study:
- To computationally design and select a thermostable mutant of tobacco 5-epi-aristolochene synthase (TEAS).
- To enable carbocation cyclization reactions at higher temperatures.
- To expand the utility of terpene synthases in industrial applications.
Main Methods:
- Computational design of TEAS variants.
- Directed evolution incorporating proteolytic digestion and protein capture for thermostability selection.
- Enzymatic assays at elevated temperatures to assess activity and stability.
Main Results:
- A thermostable mutant of TEAS was successfully engineered.
- The engineered TEAS variant retains enzymatic activity at 65°C.
- The thermostable variant denatures above 80°C, exhibiting enhanced thermal stability compared to the wild-type enzyme.
Conclusions:
- The engineered thermostable TEAS mutant demonstrates significantly improved thermal stability.
- This variant enables efficient catalysis of terpenoid synthesis at elevated temperatures.
- The findings pave the way for novel industrial applications of terpene synthases.
