Molecular Cloning, Functional Expression and Characterization of (E)-β-Farnesene Synthase from Citrus junos

• Published in: Biological & pharmaceutical bulletin 2001, Vol.24(10), pp.1171-1175
• Main Authors: MARUYAMA, Takuro, ITO, Michiho, HONDA, Gisho
• Format: Article
• Language: English
• Subjects: (E)-β-farnesene synthase; Amino Acid Sequence; Biological and medical sciences; Biotechnology; Citrus - enzymology; Citrus - genetics; Citrus junos(Rutaceae); Cloning, Molecular; Cyclization; DNA, Plant - biosynthesis; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Gas Chromatography-Mass Spectrometry; Genetic engineering; Genetic technics; Indicators and Reagents; Methods. Procedures. Technologies; Molecular cloning; Molecular Sequence Data; Pyrophosphatases - biosynthesis; Pyrophosphatases - genetics; Reverse Transcriptase Polymerase Chain Reaction; RNA, Plant - isolation & purification; RNA, Plant - metabolism; sesquiterpene biosynthesis; Sesquiterpenes - metabolism
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.24.1171

We cloned the gene of the acyclic sesquiterpene synthase, (E)-β-farnesene synthase (CJFS) from Yuzu (Citrus junos, Rutaceae). The function of CJFS was elucidated by the preparation of recombinant protein and subsequent enzyme assay. CJFS consisted of 1867 nucleotides including 1680 bp of coding sequence encoding a protein of 560 amino acids with a molecular weight of 62 kDa. The deduced amino acid sequence possessed characteristic amino acid residues, such as the DDxxD motif, which are highly conserved among terpene synthases. This is the first report of the cloning of a terpene synthase from a Rutaceous plant. A possible reaction mechanism for terpene biosynthesis is also discussed on the basis of sequence comparison of CJFS with known sesquiterpene synthase genes.