A Catalytically Competent Terpene Synthase Inferred Using Ancestral Sequence Reconstruction Strategy

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• 作者：Daniele Guzzetti ; Aurélien Lebrun ; Maeva Subileau ; Estelle Grousseau ; Eric Dubreucq ; Jullien Drone
• 刊名：ACS Catalysis
• 出版年：2016
• 出版时间：August 5, 2016
• 年：2016
• 卷：6
• 期：8
• 页码：5345-5349
• 全文大小：317K
• 年卷期：0
• ISSN：2155-5435

文摘

On the basis of bioinformatic analysis of 137 putative and characterized bacterial terpene synthases (TS), we applied ancestral reconstruction strategy to design a biosynthetic protein sequence (AncCL1). We biochemically confirmed its catalytic competence and product profile. This enzyme catalyzes not only epi-isozizaene (minor product) but also epi-zizaene (major product) formation. We compared AncCL1 with two related TS to allow a more detailed insight into the product specificity determinants. In the future, the presented method will be useful for engineering biosynthetic TS and to elucidate the function of unknown enzymes.