Hallahan D L, Lau S M, Harder P A, Smiley D W, Dawson G W, Pickett J A, Christoffersen R E, O'Keefe D P
Biochemistry and Physiology Department, AFRC Institute of Arable Crops Research, Harpenden, Herts, UK.
Biochim Biophys Acta. 1994 Sep 28;1201(1):94-100. doi: 10.1016/0304-4165(94)90156-2.
A cytochrome P-450 present in ripening avocado (Persea americana) fruit mesocarp (CYP71A1) had previously been shown to metabolize the monoterpenoids nerol and geraniol (Hallahan et al. (1992) Plant Physiol. 98, 1290-1297). Using DNA encoding CYP71A1 as a hybridization probe, we have shown by Southern analysis that a related gene is present in the catmint, Nepeta racemosa. RNA blot analysis, together with Western analysis of catmint leaf polypeptides using avocado cyt P-450 antiserum, showed that a closely related gene is expressed in catmint leaves. Cytochrome P-450 in catmint microsomes catalysed the specific hydroxylation of nerol and geraniol at C-10, whereas avocado CYP71A1, in either avocado microsomes or heterologously expressed in yeast, catalysed 2,3- or 6,7-epoxidation of these substrates. These results suggest that orthologous genes of the CYP71 family are expressed in these two plant species, but catalyse dissimilar reactions with monoterpenoid substrates.
先前已证明,成熟鳄梨(Persea americana)果实中果皮中的一种细胞色素P-450(CYP71A1)可代谢单萜类化合物橙花醇和香叶醇(Hallahan等人,(1992年)《植物生理学》98卷,1290 - 1297页)。使用编码CYP71A1的DNA作为杂交探针,我们通过Southern分析表明,荆芥(Nepeta racemosa)中存在一个相关基因。RNA印迹分析,以及使用鳄梨细胞色素P-450抗血清对荆芥叶片多肽进行的蛋白质免疫印迹分析表明,一个密切相关的基因在荆芥叶片中表达。荆芥微粒体中的细胞色素P-450催化橙花醇和香叶醇在C-10位的特异性羟基化反应,而鳄梨CYP71A1,无论是在鳄梨微粒体中还是在酵母中异源表达,都催化这些底物的2,3-或6,7-环氧化反应。这些结果表明,CYP71家族的直系同源基因在这两种植物物种中表达,但催化与单萜类底物不同的反应。