Related Experiment Video
Updated: May 1, 2026
06:51
High-throughput Screening of Carbohydrate-degrading Enzymes Using Novel Insoluble Chromogenic Substrate Assay Kits
Published on: September 20, 2016
13.6K
A high-throughput colorimetric screening assay for terpene synthase activity based on substrate consumption.
Maiko Furubayashi1, Mayu Ikezumi1, Jun Kajiwara1
1Department of Applied Chemistry and Biotechnology, Chiba University, Chiba, Japan.
Plos One
|April 1, 2014
Summary
Researchers developed a simple screening method to measure terpene synthase (TPS) activity by observing color loss in cells. This high-throughput approach aids in analyzing TPS mutations and directed evolution for new chemical structures.
Area of Science:
- Biochemistry
- Enzymology
- Metabolic Engineering
Background:
- Terpene synthases (TPS) are crucial enzymes for creating diverse terpene structures.
- Understanding TPS mechanisms and enhancing their activity in hosts is vital for terpenoid synthesis.
- Metabolic engineering and host breeding advancements drive the need for efficient TPS analysis tools.
Purpose of the Study:
- To develop a simple, high-throughput screening method for assessing cellular terpene synthase activity.
- To enable mutation analysis and directed evolution of terpene synthases.
- To explore substrate-specific screening for terpene synthases.
Main Methods:
- Developed a screening method based on substrate consumption, indicated by color loss in carotenoid pathways.
- Utilized colorimetric changes in host cells harboring carotenoid pathways to score enzyme activity.
- Explored substrate-size specificity of C30 and C40 carotenoid pathways for targeted screening.
Main Results:
- Demonstrated a high-throughput method to detect various terpene synthase and prenyltransferase gene activities.
- The method is independent of the specific terpene product generated.
- Showcased the potential for substrate-specific screening by leveraging carotenoid pathway specificities.
Conclusions:
- The developed screening method offers a facile approach for analyzing terpene synthases.
- This tool facilitates mutation analysis and directed evolution of terpene synthases.
- The method supports the engineering of terpene production in heterologous hosts.
