Anderson J D, Moore T C
Department of Botany and Plant Pathology, Oregon State University, Corvallis, Oregon 97331.
Plant Physiol. 1967 Nov;42(11):1527-34. doi: 10.1104/pp.42.11.1527.
Mevalonate-(14)C was incorporated into (-)-kaurene-(14)C in cell-free extracts of immature pea (Pisum sativum L.) seeds. The identification of (14)C-product as (-)-kaurene was based on: A) comparison with authentic (-)-kaurene on thin-layer and gas-liquid chromatography; and B) oxidation of (14)C-product and (-)-kaurene with osmium tetroxide to form the common derivative kaurane-16,17-diol. The enzyme system is heat labile and is dependent upon ATP and Mg(2+) or Mn(2-), with Mn(2+) being a more effective activator than Mg(2+). The reaction rate was proportional to enzyme concentration in reaction mixtures containing 0.45 to 1.8 mg protein n/ml, and was linear with time through 120 minutes in standard reaction mixtures. Enzyme preparations from immature seeds of tall and dwarf peas appeared to synthesize (-)-kaurene at the same rate. Synthesis of (-)-kaurene was readily inhibited by Amo-1618. (2-Chloroethyl)-trimethylammonium chloride (CCC) also inhibited (-)-kaurene synthesis; however, approximately 1000-fold higher concentrations of CCC were required to evoke the same percentages of inhibition as Amo-1618.
甲羟戊酸 -（14）C被掺入未成熟豌豆（Pisum sativum L.）种子的无细胞提取物中的（-）-贝壳杉烯 -（14）C中。将（14）C产物鉴定为（-）-贝壳杉烯基于：A）在薄层色谱和气相色谱上与纯（-）-贝壳杉烯进行比较；以及B）用四氧化锇将（14）C产物和（-）-贝壳杉烯氧化以形成共同衍生物贝壳杉烷 - 16,17 - 二醇。该酶系统对热不稳定，并且依赖于ATP和Mg（2+）或Mn（2+），其中Mn（2+）是比Mg（2+）更有效的激活剂。在含有0.45至1.8mg蛋白质/ nml的反应混合物中，反应速率与酶浓度成正比，并且在标准反应混合物中120分钟内与时间呈线性关系。来自高豌豆和矮豌豆未成熟种子的酶制剂似乎以相同的速率合成（-）-贝壳杉烯。（-）-贝壳杉烯的合成很容易被Amo - 1618抑制。（2 - 氯乙基）-三甲基氯化铵（CCC）也抑制（-）-贝壳杉烯的合成；然而，需要大约高1000倍浓度的CCC才能产生与Amo - 1618相同百分比的抑制作用。
