Basyuni Mohammad, Oku Hirosuke, Tsujimoto Etsuko, Baba Shigeyuki
United Graduate School of Agricultural Sciences, Kagoshima University, Korimoto, Kagoshima, Japan.
Biosci Biotechnol Biochem. 2007 Jul;71(7):1788-92. doi: 10.1271/bbb.70113. Epub 2007 Jul 7.
To obtain cDNAs encoding oxidosqualene cyclase (OSC), we cloned two cDNAs, KcCAS and RsCAS, from roots of Kandelia candel (L.) Druce and leaves of Rhizophora stylosa Griff. by homology based PCR method respectively. The deduced amino acid sequences of both OSCs showed 82% homology to cycloartenol synthases from Lotus japonicus (OSC5) and Ricinus cummunis (RcCAS), suggesting that these are cycloartenol synthases of K. candel and R. stylosa. The genes obtained were expressed in a lanosterol synthase deficient Saccharomyces cerevisiae (ERG7) strain, GIL77. GC-MS analysis identified the accumulated reaction product in the yeast transformant to be cycloartenol, indicating that both KcCAS and RsCAS encode cycloartenol synthase.
为了获得编码氧化鲨烯环化酶(OSC)的cDNA,我们分别通过基于同源性的PCR方法,从秋茄(Kandelia candel (L.) Druce)的根和红海榄(Rhizophora stylosa Griff.)的叶中克隆了两个cDNA,即KcCAS和RsCAS。两种OSC的推导氨基酸序列与来自日本百脉根(Lotus japonicus)(OSC5)和蓖麻(Ricinus cummunis)(RcCAS)的环阿屯醇合酶显示出82%的同源性,这表明它们是秋茄和红海榄的环阿屯醇合酶。将获得的基因在缺乏羊毛甾醇合酶的酿酒酵母(Saccharomyces cerevisiae)(ERG7)菌株GIL77中表达。气相色谱-质谱分析确定酵母转化体中积累的反应产物为环阿屯醇,表明KcCAS和RsCAS均编码环阿屯醇合酶。