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Seven cDNA clones encoding terpene synthases (TPSs), their structures closely related to each other, were isolated from the flower of  Camellia hiemalis  (‘Kantsubaki’). Their putative TPS proteins were phylogenetically positioned in a sole clade with the TPSs of other  Camellia  species. The obtained  Tps  genes, one of which was designated  ChTps1  ( ChTps1a ), were introduced into mevalonate-pathway-engineered  Escherichia coli , which carried the genes for utilizing acetoacetate as a substrate, and cultured in a medium including lithium acetoacetate. Volatile products generated in the  E .  coli  cells transformed with  ChTps1  were purified from the cell suspension culture, and analyzed by NMR. Consequently, the predominant product with ChTPS1 was identified as valerianol, indicating that the  ChTps1  gene codes for valerianol synthase. This is the first report on a gene that can mediate the synthesis of valerianol. We next synthesized a  Tps  ortholog encoding ChTPS1variant R477H (named CsiTPS8), whose sequence had been isolated from a tea tree ( Camellia sinensis ), carried out similar culture experiment with the  E .  coli  transformant including  CsiTps8 , and consequently found valerianol production equally. Furthermore, GC-MS analysis of several teas revealed that valerianol had been an unknown ingredient in green tea and black tea.

Identification of novel sesquiterpene synthase genes that mediate the biosynthesis of valerianol, which was an unknown ingredient of tea