Isolation and functional characterization of a beta-eudesmol synthase, a new sesquiterpene synthase from Zingiber zerumbet Smith.
In this paper, we have identified a new sesquiterpene synthase gene (ZSS2) from Zingiber zerumbet Smith. Functional expression of ZSS2 in Escherichia coli and in vitro enzyme assay showed that the encoded enzyme catalyzed the formation of beta-eudesmol and five additional by-products. Quantitative RT-PCR analysis revealed that ZSS2 transcript accumulation in rhizomes has strong seasonal variations. To further confirm the enzyme activity of ZSS2 and to assess the potential for metabolic engineering of beta-eudesmol production, we introduced a gene cluster encoding six enzymes of the mevalonate pathway into E. coli and coexpressed it with ZSS2. When supplemented with mevalonate, the engineered E. coli produced a similar sesquiterpene profile to that produced in the in vitro enzyme assay, and the yield of beta-eudesmol reached 100 mg/L.
Yu
Fengnian
F
Department of Bioscience, Graduate School of Agriculture, Kinki University, Nakamachi, Nara, Japan.
Harada
Hisashi
H
Yamasaki
Kazuhisa
K
Okamoto
Sho
S
Hirase
Souta
S
Tanaka
Yasuo
Y
Misawa
Norihiko
N
Utsumi
Ryutaro
R
D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli; D008401:Gas Chromatography-Mass Spectrometry; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D005818:Genetic Engineering; D008798:Mevalonic Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D010938:Plant Oils / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D027343:Rhizome / Q000737:chemistry; D012621:Seasons; D012717:Sesquiterpenes / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D045787:Sesquiterpenes, Eudesmane / Q000737:chemistry / Q000378:metabolism*; D030521:Zingiberaceae / Q000201:enzymology* / Q000235:genetics
D010938:Plant Oils; D010940:Plant Proteins; D011106:Polyisoprenyl Phosphates; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012717:Sesquiterpenes; D045787:Sesquiterpenes, Eudesmane; C051082:beta-eudesmol; C004808:farnesyl pyrophosphate; D019883:Alkyl and Aryl Transferases; D008798:Mevalonic Acid