Terpenoids have many biological functions and a comprehensive range of applications. Here, we cloned two monoterpene synthase genes, Tc-αpin/teo and Tc-teo, from Taiwania cryptomerioides. The enzymes encoded by these genes shared 97 % amino acid sequences similarity but had different terpene product profiles. Using structural modeling and site-directed mutagenesis, we successfully identified three plasticity residues around the active site of Tc-αPIN/TEO, namely Y327, Y429 and Y575 that are involved in secondary cyclization. The mutants in which the phenolic residues were replaced with phenylalanines seemed to lose their preference for α-pinene synthesis, indicating that the tyrosine hydroxyl groups at these sites were necessary for the formation of bicyclic terpene products.
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We thank Prof. Shang-Tzen Chang (School of Forestry and Resource Conservation, National Taiwan University) for providing the GC-MS apparatus. The financial assistance from the National Science Council (NSC 102-2628-B-002-012-MY3) is gratefully acknowledged.
Communicated by R. Sederoff
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Hsu, L., Chu, F. Plasticity residues involved in secondary cyclization of terpene synthesis in Taiwania cryptomerioides . Tree Genetics & Genomes 11, 796 (2015). https://doi.org/10.1007/s11295-014-0796-z
- Monoterpene synthase
- Plasticity residues
- Protein engineering
- Secondary cyclization
- Site-directed mutagenesis