Molecular cloning and functional expression of triterpene synthases from pea (Pisum sativum)
作者： Masayo Morita , Masaaki Shibuya , Tetsuo Kushiro , Kazuo Masuda , Yutaka Ebizuka
DOI: 10.1046/J.1432-1327.2000.01357.X
关键词: Lupeol synthase 、 Heterologous expression 、 Biochemistry 、 Triterpene 、 Stereochemistry 、 Expression vector 、 Sativum 、 Molecular cloning 、 Amyrin 、 Biology 、 Oleanane
摘要: Ursane type triterpene is one of the most widespread aglycones found in plants, together with oleanane type, and these two types often occur same plant. Pisum sativum known to produce both triterpenes. Homology based PCRs degenerate primers designed from conserved sequences beta-amyrin synthases have resulted cloning synthase cDNAs immature seeds P. sativum. They show high sequence identities each other (78%) also (70-90%). ORFs full-length clones named as PSY (2277 bp, codes for 759 amino acids) PSM (2295 765 were ligated into yeast expression vector pYES2 under control GAL1 promoter. Heterologous revealed be a synthase. Surprisingly, however, turned out novel mixed amyrin producing alpha- beta-amyrin. Several minor triterpenes identified byproducts. The presence such multifunctional would account co-occurence ursane plants.
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