tailieunhanh - Báo cáo khoa học: An unusual plant triterpene synthase with predominant a-amyrin-producing activity identified by characterizing oxidosqualene cyclases from Malus · domestica

The pentacyclic triterpenes, in particular ursolic acid and oleanolic acid and their derivatives, exist abundantly in the plant kingdom, where they are well known for their anti-inflammatory, antitumour and antimicrobial properties. | IFEBS Journal An unusual plant triterpene synthase with predominant a-amyrin-producing activity identified by characterizing oxidosqualene cyclases from Malus X domestica Cyril Brendolise1 Yar-Khing Yauk1 Ellen D. Eberhard1 Mindy Wang1 David Chagne1 Christelle Andre1 David R. Greenwood1 2 and Lesley L. Beuning1 f 1 The New Zealand Institute for Plant and Food Research Limited Plant Food Research Auckland New Zealand 2 Schoolof BiologicalSciences University of Auckland New Zealand Keywords apple ursolic acid triterpene synthase a-amyrin b-amyrin Correspondence D. R. Greenwood Mt Albert Research Centre Plant Food Research Private Bag 92 169 Auckland 1142 New Zealand Fax 64 9 925 7001 Tel 64 9 925 7147 E-mail Received 9 February 2009 revised 2 May 2011 accepted 10 May 2011 doi The pentacyclic triterpenes in particular ursolic acid and oleanolic acid and their derivatives exist abundantly in the plant kingdom where they are well known for their anti-inflammatory antitumour and antimicrobial properties. a-Amyrin and b-amyrin are the precursors of ursolic and olean-olic acids respectively formed by concerted cyclization of squalene epoxide by a complex synthase reaction. We identified three full-length expressed sequence tag sequences in cDNA libraries constructed from apple Malus X domestica Royal Gala that were likely to encode triterpene synthases. Two of these expressed sequence tag sequences were essentially identical 99 amino acid similarity MdOSCl and MdOSC3 . MdOSC1 and MdOSC2 were expressed by transient expression in Nicotiana benthamiana leaves and by expression in the yeast Pichia methanolica. The resulting products were analysed by GC and GC-MS. MdOSC1 was shown to be a mixed amyrin synthase a 5 1 ratio of a-amyrin to b-amyrin . MdOSC1 is the only triterpene synthase so far identified in which the level of a-amyrin produced is 80 of the total product and is therefore primarily an a-amyrin .

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