Identification of Vitis vinifera (-)-alpha-terpineol synthase by in silico screening of full-length cDNA ESTs and functional characterization of recombinant terpene synthase.
The flavour and aroma of certain Vitis vinifera grape varieties is dominated by volatile terpenes and small volatile aldehydes. Monoterpenes contribute to the final grape and wine aroma and flavour in form of free volatiles and as glycoside conjugates of monoterpene alcohols. Typical monoterpenol components of the cultivar Gewürztraminer and other aroma-rich grape varieties are linalool, geraniol, nerol, citronellol, and alpha-terpineol. In a functional genomics effort to identify genes for the formation of monoterpene alcohols in V. vinifera, a database of full-length cDNA sequences was screened in silico and yielded two clones for putative monoterpene synthases. The gene products were functionally characterized by expression in Escherichia coli, in vitro enzyme assay and gas chromatography-mass spectrometry (GC-MS) product identification as multi-product (-)-alpha-terpineol synthases.
Martin
Diane M
DM
Biotechnology Laboratory, University of British Columbia, 6174 University Boulevard, Rm. 237, Vancouver, BC, Canada V6T 1Z3.
Bohlmann
Jörg
J
D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000081005:Cyclohexane Monoterpenes; D053138:Cyclohexenes; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D020224:Expressed Sequence Tags; D008401:Gas Chromatography-Mass Spectrometry; D008969:Molecular Sequence Data; D039821:Monoterpenes / Q000737:chemistry / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013237:Stereoisomerism; D027843:Vitis / Q000201:enzymology* / Q000235:genetics
D000081005:Cyclohexane Monoterpenes; D053138:Cyclohexenes; D018076:DNA, Complementary; D039821:Monoterpenes; D011994:Recombinant Proteins; C016775:alpha-terpineol; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase