Disch A, Schwender J, Müller C, Lichtenthaler H K, Rohmer M
Université Louis Pasteur/CNRS, Institut Le Bel, 4 rue Blaise Pascal, 67070 Strasbourg Cedex, France.
Biochem J. 1998 Jul 15;333 ( Pt 2)(Pt 2):381-8. doi: 10.1042/bj3330381.
Isopentenyl diphosphate, the universal isoprenoid precursor, can be produced by two different biosynthetic routes: either via the acetate/mevalonate (MVA) pathway, or via the more recently identified MVA-independent glyceraldehyde phosphate/pyruvate pathway. These two pathways are easily differentiated by incorporation of [1-13C]glucose and analysis of the resulting labelling patterns found in the isoprenoids. This method was successfully applied to several unicellular algae raised under heterotrophic growth conditions and allowed for the identification of the pathways that were utilized for isoprenoid biosynthesis. All isoprenoids examined (sterols, phytol, carotenoids) of the green algae Chlorella fusca and Chlamydomonas reinhardtii were synthesized via the GAP/pyruvate pathway, as in another previously investigated green alga, Scenedesmus obliquus, which was also shown in this study to synthesize ubiquinone by the same MVA-independent route. In the red alga Cyanidium caldarium and in the Chrysophyte Ochromonas danica a clear dichotomy was observed: as in higher plants, sterols were formed via the MVA route, whereas chloroplast isoprenoids (phytol in Cy. caldarium and O. danica and beta-carotene in O. danica) were synthesized via the GAP/pyruvate route. In contrast, the Euglenophyte Euglena gracilis synthesized ergosterol, as well as phytol, via the acetate/MVA route. Similar feeding experiments were performed with the cyanobacterium Synechocystis PCC 6714 using [1-13C]- and [6-13C]-glucose. The two isoprenoids examined, phytol and beta-carotene, were shown to have the typical labelling pattern derived from the GAP/pyruvate route.
异戊烯基二磷酸是通用的类异戊二烯前体,可通过两条不同的生物合成途径产生:要么通过乙酸/甲羟戊酸(MVA)途径,要么通过最近发现的不依赖MVA的甘油醛磷酸/丙酮酸途径。通过掺入[1-13C]葡萄糖并分析类异戊二烯中产生的标记模式,可以很容易地区分这两条途径。该方法已成功应用于在异养生长条件下培养的几种单细胞藻类,并有助于确定用于类异戊二烯生物合成的途径。绿藻小球藻和莱茵衣藻中检测的所有类异戊二烯(甾醇、叶绿醇、类胡萝卜素)均通过甘油醛磷酸/丙酮酸途径合成,就像另一种先前研究过的绿藻斜生栅藻一样,本研究还表明该绿藻通过相同的不依赖MVA的途径合成泛醌。在红藻嗜热栖热菌和金藻丹麦赭球藻中观察到明显的二分法:与高等植物一样,甾醇通过MVA途径形成,而叶绿体类异戊二烯(嗜热栖热菌和丹麦赭球藻中的叶绿醇以及丹麦赭球藻中的β-胡萝卜素)通过甘油醛磷酸/丙酮酸途径合成。相比之下,裸藻纤细裸藻通过乙酸/MVA途径合成麦角甾醇以及叶绿醇。使用[1-13C]-和[6-13C]-葡萄糖对蓝藻集胞藻PCC 6714进行了类似的饲喂实验。检测的两种类异戊二烯,叶绿醇和β-胡萝卜素,显示出源自甘油醛磷酸/丙酮酸途径的典型标记模式。