Takahashi Shunji, Yeo Yunsoo, Greenhagen Bryan T, McMullin Tom, Song Linsheng, Maurina-Brunker Julie, Rosson Reinhardt, Noel Joseph P, Chappell Joe
Department of Plant and Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312, USA.
Biotechnol Bioeng. 2007 May 1;97(1):170-81. doi: 10.1002/bit.21216.
Terpenes are structurally diverse compounds that are of interest because of their biological activities and industrial value. These compounds consist of chirally rich hydrocarbon backbones derived from terpene synthases, which are subsequently decorated with hydroxyl substituents catalyzed by terpene hydroxylases. Availability of these compounds is, however, limited by intractable synthetic means and because they are produced in low amounts and as complex mixtures by natural sources. We engineered yeast for sesquiterpene accumulation by introducing genetic modifications that enable the yeast to accumulate high levels of the key intermediate farnesyl diphosphate (FPP). Co-expression of terpene synthase genes diverted the enlarged FPP pool to greater than 80 mg/L of sesquiterpene. Efficient coupling of terpene production with hydroxylation was also demonstrated by coordinate expression of terpene hydroxylase activity, yielding 50 mg/L each of hydrocarbon and hydroxylated products. These yeast now provide a convenient format for investigating catalytic coupling between terpene synthases and hydroxylases, as well as a platform for the industrial production of high value, single-entity and stereochemically unique terpenes.
萜类化合物是结构多样的化合物，因其生物活性和工业价值而备受关注。这些化合物由源自萜烯合酶的富含手性的烃骨架组成，随后由萜烯羟化酶催化用羟基取代基进行修饰。然而，这些化合物的可得性受到难以处理的合成方法的限制，并且因为它们由天然来源以低产量和复杂混合物的形式产生。我们通过引入基因修饰来改造酵母以积累倍半萜，这些修饰使酵母能够积累高水平的关键中间体法呢基二磷酸（FPP）。萜烯合酶基因的共表达将扩大的FPP库转化为超过80 mg/L的倍半萜。通过萜烯羟化酶活性的协同表达也证明了萜类化合物生产与羟基化的有效偶联，产生了各50 mg/L的烃类和羟基化产物。这些酵母现在为研究萜烯合酶和羟化酶之间的催化偶联提供了一种便利的形式，同时也是用于工业生产高价值、单一实体和立体化学独特的萜类化合物的平台。
