Identification of a product specific β-amyrin synthase from Arabidopsis thaliana |
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Authors: | Masaaki Shibuya Yuji Katsube Miyuki Otsuka Hong Zhang Pimpimon Tansakul Ting Xiang Yutaka Ebizuka |
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Affiliation: | 1. The National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, PR China;2. Department of Plant Science, Plant Genomics and Breeding Institute, Research Institute for Agriculture and Life Science, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Republic of Korea;3. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China |
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Abstract: | Triterpene skeletons are produced by oxidosqualene cyclases (OSCs). The genome sequencing of Arabidopsis thaliana revealed the presence of thirteen OSC homologous genes including At1g78950, which has been revised recently as two independent ORFs, namely At1g78950 and At1g78955. The cDNA corresponding to the revised At1g78950 was obtained by RT-PCR, ligated into Saccharomyces cerevisiae expression vector pYES2, and expressed in a lanosterol synthase deficient S. cerevisiae strain. LC-MS and NMR analyses of the accumulated product in the host cells showed that the product of At1g78950 is β-amyrin, indicating that At1g78950 encodes a β-amyrin synthase (EC 5.4.99.-). |
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