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Functional characterization and substrate promiscuity of sesquiterpene synthases from Tripterygium wilfordii.
Tong, Yuru; Hu, Tianyuan; Tu, Lichan; Chen, Kang; Liu, Tiezheng; Su, Ping; Song, Yadi; Liu, Yuan; Huang, Luqi; Gao, Wei.
Affiliation
  • Tong Y; School of Pharmaceutical Sciences, Capital Medical University, Beijing 100069, PR China. Electronic address: tongyuru@ccmu.edu.cn.
  • Hu T; College of Pharmacy, School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang 311121, PR China.
  • Tu L; School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, PR China.
  • Chen K; National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, PR China.
  • Liu T; National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, PR China.
  • Su P; Department of Chemistry, the Scripps Research Institute, Jupiter, FL 33458, USA.
  • Song Y; Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, PR China.
  • Liu Y; School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, PR China.
  • Huang L; National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, PR China. Electronic address: huangluqi01@126.com.
  • Gao W; Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, PR China; School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, PR China. Electronic address: weigao@ccmu.edu.cn.
Int J Biol Macromol ; 185: 949-958, 2021 Aug 31.
Article in En | MEDLINE | ID: mdl-34237366
Acyclic terpenes, commonly found in plants, are of high physiological importance and commercial value, and their diversity was controlled by different terpene synthases. During the screen of sesquiterpene synthases from Tripterygium wilfordii, we observed that Ses-TwTPS1-1 and Ses-TwTPS2 promiscuously accepted GPP, FPP, and GGPP to produce corresponding terpene alcohols (linalool/nerolidol/geranyllinalool). The Ses-TwTPS1-2, Ses-TwTPS3, and Ses-TwTPS4 also showed unusual substrate promiscuity by catalyzing GGPP or GPP in addition to FPP as substrate. Furthermore, key residues for the generation of diterpene product, (E, E)-geranyllinalool, were screened depending on mutagenesis studies. The functional analysis of Ses-TwTPS1-1:V199I and Ses-TwTPS1-2:I199V showed that Val in 199 site assisted the produce of diterpene product geranyllinalool by enzyme mutation studies, which indicated that subtle differences away from the active site could alter the product outcome. Moreover, an engineered sesquiterpene high-yielding yeast that produced 162 mg/L nerolidol in shake flask conditions was constructed to quickly identify the function of sesquiterpene synthases in vivo and develop potential applications in microbial fermentation. Our functional characterization of acyclic sesquiterpene synthases will give some insights into the substrate promiscuity of diverse acyclic terpene synthases and provide key residues for expanding the product portfolio.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Alkyl and Aryl Transferases / Tripterygium Language: En Journal: Int J Biol Macromol Year: 2021 Document type: Article Country of publication: Netherlands

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Alkyl and Aryl Transferases / Tripterygium Language: En Journal: Int J Biol Macromol Year: 2021 Document type: Article Country of publication: Netherlands