Downloads: 0

Files in This Item:
This article will be available after a certain embargo period.
Please see the "Rights" information in item metadata display about embargo date.
Title: Characterization of Class IB Terpene Synthase: The First Crystal Structure Bound with a Substrate Surrogate
Authors: Stepanova, Rafaella
Inagi, Hayato
Sugawara, Kei
Asada, Kazuya
Nishi, Tomoyuki
Ueda, Daijiro
Yasuno, Yoko
Shinada, Tetsuro
Miki, Kunio
Fujihashi, Masahiro  kyouindb  KAKEN_id  orcid https://orcid.org/0000-0002-6882-9697 (unconfirmed)
Sato, Tsutomu
Author's alias: 稲木, 隼人
菅原, 啓
西, 智之
上田, 大次郎
保野, 陽子
品田, 哲郎
三木, 邦夫
藤橋, 雅宏
佐藤, 努
Issue Date: 19-Jun-2020
Publisher: American Chemical Society (ACS)
Journal title: ACS Chemical Biology
Volume: 15
Issue: 6
Start page: 1517
End page: 1525
Abstract: Terpene synthases (TS) are classified into two broad types, Class I and II, based on the chemical strategy for initial carbocation formation and motif sequences of the catalytic site. We have recently identified a new class of enzymes, Class IB, showing the acceptability of long (C₂₀–C₃₅) prenyl-diphosphates as substrates and no amino acid sequence homology with known TS. Conversion of long prenyl-diphosphates such as heptaprenyl-diphosphate (C₃₅) is unusual and has never been reported for Class I and II enzymes. Therefore, the characterization of Class IB enzymes is crucial to understand the reaction mechanism of the extensive terpene synthesis. Here, we report the crystal structure bound with a substrate surrogate and biochemical analysis of a Class IB TS, using the enzyme from Bacillus alcalophilus (BalTS). The structure analysis revealed that the diphosphate part of the substrate is located around the two characteristic Asp-rich motifs, and the hydrophobic tail is accommodated in a unique hydrophobic long tunnel, where the C₃₅ prenyl-diphosphate, the longest substrate of BalTS, can be accepted. Biochemical analyses of BalTS showed that the enzymatic property, such as Mg^2⁺ dependency, is similar to those of Class I enzymes. In addition, a new cyclic terpene was identified from BalTS reaction products. Mutational analysis revealed that five of the six Asp residues in the Asp-rich motifs and two His residues are essential for the formation of the cyclic skeleton. These results provided a clue to consider the application of the unusual large terpene synthesis by Class IB enzymes.
Rights: This document is the Accepted Manuscript version of a Published Work that appeared in final form in 'ACS Chemical Biology', copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acschembio.0c00145
The full-text file will be made open to the public on 31 March 2021 in accordance with publisher's 'Terms and Conditions for Self-Archiving'
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/2433/255583
DOI(Published Version): 10.1021/acschembio.0c00145
PubMed ID: 32227910
Appears in Collections:Journal Articles

Show full item record

Export to RefWorks


Export Format: 


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.