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Title: A domain swapping approach to elucidate differential regiospecific hydroxylation by geraniol and linalool synthases from perilla.
Authors: Sato-Masumoto, Naoko
Ito, Michiho  kyouindb  KAKEN_id
Author's alias: 佐藤(増本) , 直子
伊藤, 美千穂
Keywords: Perilla
Labiatae
Geraniol synthase
Linalool synthase
Domain swapping
Site-directed mutagenesis
Chimeric enzymes
Regiospecific hydroxylation
Essential oil
Issue Date: Jun-2014
Publisher: Elsevier Ltd.
Journal title: Phytochemistry
Volume: 102
Start page: 46
End page: 54
Abstract: Geraniol and linalool are acyclic monoterpenes found in plant essential oils that have attracted much attention for their commercial use and in pharmaceutical studies. They are synthesized from geranyl diphosphate (GDP) by geraniol and linalool synthases, respectively. Both synthases are very similar at the amino acid level and share the same substrate; however, the position of the GDP to which they introduce hydroxyl groups is different. In this study, the mechanisms underlying the regiospecific hydroxylation of geraniol and linalool synthases were investigated using a domain swapping approach and site-directed mutagenesis in perilla. Sequences of the synthases were divided into ten domains (domains I to IV-4), and each corresponding domain was exchanged between both enzymes. It was shown that different regions were important for the formation of geraniol and linalool, namely, domains IV-1 and -4 for geraniol, and domains III-b, III-d, and IV-4 for linalool. These results suggested that the conformation of carbocation intermediates and their electron localization were seemingly to be different between geraniol and linalool synthases. Further, five amino acids in domain IV-4 were apparently indispensable for the formation of geraniol and linalool. According to three-dimensional structural models of the synthases, these five residues seemed to be responsible for the different spatial arrangement of the amino acid at H524 in the case of geraniol synthase, while N526 is the corresponding residue in linalool synthase. These results suggested that the side-chains of these five amino acids, in combination with several relevant domains, localized the positive charge in the carbocation intermediate to determine the position of the introduced hydroxyl group.
Rights: © 2014 Elsevier Ltd.
This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。
URI: http://hdl.handle.net/2433/187741
DOI(Published Version): 10.1016/j.phytochem.2014.03.020
PubMed ID: 24725978
Appears in Collections:Journal Articles

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