Characterization of a sesquiterpene synthase and a short-chain dehydrogenase in zerumbone biosynthesis and the applications in engineered Saccharomyces cerevisiae

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Publicat a:Frontiers in Plant Science vol. 16 (Oct 2025), p. 1635141-1635153
Autor principal: Xu, Mengdie
Altres autors: Xia, Yimeng, Fang, Gaowei, Li, Tangli, Ma, Jing, Li, Dengyu, Qiuhui Wei, Tu, Lichan, Yin, Xiaopu, Engineering Laboratory of Development and Application of Traditional Chinese Medicines, Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, China"], Hu, Tianyuan
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Frontiers Media SA
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022 |a 1664-462X 
024 7 |a 10.3389/fpls.2025.1635141  |2 doi 
035 |a 3273797852 
045 2 |b d20251001  |b d20251031 
100 1 |a Xu, Mengdie  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China 
245 1 |a Characterization of a sesquiterpene synthase and a short-chain dehydrogenase in zerumbone biosynthesis and the applications in engineered Saccharomyces cerevisiae 
260 |b Frontiers Media SA  |c Oct 2025 
513 |a Journal Article 
520 3 |a IntroductionZerumbone is a pharmacologically active sesquiterpenoid with limited availability. This study aims to elucidate its biosynthetic pathway in Curcuma wenyujin by identifying and characterizing the key enzymes responsible for its production.MethodsCandidate genes were selected via transcriptome analysis and phylogenetics. CwTPS8 and CwSDR1 were cloned and functionally characterized using in vitro enzyme assays and heterologous expression in engineered Saccharomyces cerevisiae . Molecular docking and site-directed mutagenesis were applied to investigate the catalytic mechanism of CwTPS8.ResultsCwTPS8 was identified as a multifunctional sesquiterpene synthase that catalyzes the formation of α-humulene (a key zerumbone precursor) and β-caryophyllene as main products, along with several minor sesquiterpenes. Mutagenesis studies identified critical residues (e.g., Thr437, Cys436) that significantly shift product specificity toward α-humulene. CwSDR1 was characterized as a short-chain dehydrogenase that efficiently oxidizes 8-hydroxy-α-humulene to zerumbone. A de novo biosynthetic pathway was reconstructed in yeast, resulting in zerumbone production at 0.50 μg/L.DiscussionThis study expands the genetic toolkit for zerumbone biosynthesis and provides insights into enzyme engineering and metabolic engineering strategies to enhance production. Limitations in precursor supply and catalytic efficiency highlight areas for future optimization. 
653 |a Transcriptomes 
653 |a Biosynthesis 
653 |a Software 
653 |a Caryophyllene 
653 |a Metabolic engineering 
653 |a E coli 
653 |a Metabolism 
653 |a Yeast 
653 |a Genes 
653 |a Dehydrogenases 
653 |a Factories 
653 |a Physicochemical properties 
653 |a Sesquiterpenes 
653 |a Dehydrogenase 
653 |a Oxidation 
653 |a Molecular docking 
653 |a Site-directed mutagenesis 
653 |a Cloning 
653 |a Zerumbone 
653 |a DNA polymerase 
653 |a Precursors 
653 |a Humulene 
653 |a Sesquiterpene synthase 
653 |a Enzymes 
653 |a Saccharomyces cerevisiae 
653 |a Environmental 
700 1 |a Xia, Yimeng  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China 
700 1 |a Fang, Gaowei  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China 
700 1 |a Li, Tangli  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China 
700 1 |a Ma, Jing  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China 
700 1 |a Li, Dengyu  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China 
700 1 |a Qiuhui Wei  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China, School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China, Xinchang Pharmaceutical Factory, Zhejiang Medicine Co., Ltd., Shaoxing, Zhejiang, China 
700 1 |a Tu, Lichan  |u Department of Pharmacy, School of Medicine, Hangzhou City University, Hangzhou, Zhejiang, China 
700 1 |a Yin, Xiaopu  |u Xiaopu Yin [AFFILIATION="School of Pharmacy, Hangzhou Normal University, Hangzhou, China, Key Laboratory of Elemene Class Anti-Cancer Chinese Medicines 
700 1 |a Engineering Laboratory of Development and Application of Traditional Chinese Medicines 
700 1 |a Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, China"] 
700 1 |a Hu, Tianyuan  |u School of Pharmacy, Hangzhou Normal University, Hangzhou, China, School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China, Xinchang Pharmaceutical Factory, Zhejiang Medicine Co., Ltd., Shaoxing, Zhejiang, China, State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, China Academy of Chinese Medical Sciences, Beijing, China 
773 0 |t Frontiers in Plant Science  |g vol. 16 (Oct 2025), p. 1635141-1635153 
786 0 |d ProQuest  |t Agriculture Science Database 
856 4 1 |3 Citation/Abstract  |u https://www.proquest.com/docview/3273797852/abstract/embedded/L8HZQI7Z43R0LA5T?source=fedsrch 
856 4 0 |3 Full Text  |u https://www.proquest.com/docview/3273797852/fulltext/embedded/L8HZQI7Z43R0LA5T?source=fedsrch 
856 4 0 |3 Full Text - PDF  |u https://www.proquest.com/docview/3273797852/fulltextPDF/embedded/L8HZQI7Z43R0LA5T?source=fedsrch