Dehal S S, Croteau R
Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.
Arch Biochem Biophys. 1988 Mar;261(2):346-56. doi: 10.1016/0003-9861(88)90350-5.
Humulene cyclase and caryophyllene cyclase, two enzymes which catalyze the cyclization of farnesyl pyrophosphate to the respective sesquiterpene olefins, have been partially purified from the supernatant fraction of a sage (Salvia officinalis) leaf epidermis extract and separated from each other by a combination of hydrophobic interaction, gel filtration, and ion-exchange chromatography. The molecular weight of both cyclases was estimated by gel filtration to be 57,000 and both cyclases exhibited a pH optimum of 6.5 and preferred Mg2+ (Km approximately 1.5 mM) as the required divalent metal cation. Both enzymes possessed a Km of about 1.7 microM for farnesyl pyrophosphate, were strongly inhibited by p-hydroxymercuribenzoate, and exhibited comparable sensitivities to a variety of other potential inhibitors. The properties of the two sesquiterpene olefin cyclases, which are the first from a higher plant source to be examined in detail, were very similar to each other and to other monoterpene, sesquiterpene, and diterpene cyclases previously described.
葎草烯环化酶和石竹烯环化酶这两种酶可催化法呢基焦磷酸环化生成各自的倍半萜烯烃,它们已从鼠尾草(丹参)叶表皮提取物的上清液部分得到部分纯化,并通过疏水相互作用、凝胶过滤和离子交换色谱相结合的方法彼此分离。通过凝胶过滤估计两种环化酶的分子量均为57,000,并且两种环化酶的最适pH均为6.5,并且更倾向于Mg2+(Km约为1.5 mM)作为所需的二价金属阳离子。两种酶对法呢基焦磷酸的Km约为1.7 microM,均受到对羟基汞苯甲酸的强烈抑制,并且对多种其他潜在抑制剂表现出相当的敏感性。这两种倍半萜烯烃环化酶是首次从高等植物来源进行详细研究的,它们的性质彼此非常相似,并且与先前描述的其他单萜、倍半萜和二萜环化酶相似。