Journal
BIOLOGICAL & PHARMACEUTICAL BULLETIN
Volume 32, Issue 5, Pages 818-824Publisher
PHARMACEUTICAL SOC JAPAN
DOI: 10.1248/bpb.32.818
Keywords
Gentiana straminea; beta-amyrin synthase; methyl jasmonate; Pichia pastoris; triterpenoid
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Funding
- National Natural Science Foundation of China [30771116, 30470154]
- Chinese Ministry of Education New Century Training Program Foundation [NCET-05-0581]
- Chinese Natural Education Ministry Doctor Station Foundation [20050422015]
- Excellent Youth Foundation of Shandong Province of China [JQ200810]
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Phytosterols and triterpenes are synthesized by oxidosqualene cyclases (OSCs) via the isoprenoid pathway. Here, GsAS1-a full-length beta-amyrin synthase cDNA isolated from Gentiana straminea MAXIM.-was characterized. Its open reading frame consists of 2268 bp, predicted to encode a 756 residue protein containing four QW and one Asp-Cys-Thr-Ala-Glu (DCTAE) motifs, which are both well conserved among known triterpene synthases. The deduced GsAS1 peptide sequence shares 76.2% homology with that of Panax ginseng beta-amyrin synthase. A phylogenetic analysis showed that GsAS1 is closely related to other plant OSCs, and particularly to the beta-amyrin synthases. When the GsAS1 sequence was heterologously expressed in Escherichia coli, an 88 kDa gene product was produced, and this reacted with the appropriate antibody. The sequence was also heterologously expressed in the Pichia pastoris yeast. GsAS1 is expressed in a tissue-specific manner, with its expression in the leaf being ca. 4.5-fold than that in the root, and nearly three-fold than that in the stem. GsAS1 expression was up-regulated by treatment with methyl jasmonate (MeJA) over a period from 6 h to 10 d post treatment. The accumulation oleanolic acid increased after induction by MeJA.
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