红景天甙生物合成途径:酪醇合成的起始反应及其糖基化

Salidroside biosynthesis pathway:the initial reaction and glycosylation of tyrosol

红景天甙(Salidroside)生源途径分子机制的解析是利用基因工程、代谢工程技术合成目标化合物的基础。糖基化是红景天甙生物合成的最后一步反应。在前期工作中,本课题组率先报道了与红景天甙生物合成相关的3个尿苷二磷酸葡萄糖基转移酶(UGTs)基因,在体外酶学性质研究的基础上,利用根癌农杆菌和发根农杆菌介导分别建立了相关转基因体系,鉴别了红景天甙生物合成最适UGT及植物和毛状根生物反应器系统合成红景天甙的效率差异;酪醇(Tyrosol)是红景天甙糖基化反应的甙元底物分子,其具体的代谢通路及其调控机制仍不明确。针对酪醇生物合成来源主要存在两种观点:一是酪醇可能来自于苯丙烷代谢途径产生的4-香豆酸,该途径起源于苯丙氨酸;二是生物碱代谢途径的中间产物酪胺可能是酪醇生物合成的前体,该途径则起源于酪氨酸。在后续工作中,否定了酪醇来源于苯丙烷代谢途径的可能性,进一步的工作证实酪氨酸脱羧酶(TyrDC)在酪醇生物合成的起始反应中担负着重要功能,酪醇作为一种苯乙烷类化合物衍生物,其生物合成来源于生物碱代谢途径。

Salidroside,the 8-O-β-D-glucoside of tyrosol,is a novel adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A.Bor.Due to the scarcity of R.sachalinensis and its low yield of salidroside,there is great interest in enhancing the production of salidroside by biotechnological process.Glucosylation of tyrosol is thought to be the final step in salidroside biosynthesis.In our related works,three UGT clones were isolated from the roots and the cultured cells.Our intention was to combine the catalytic specificity of these UGTs in vitro in order to change the level of salidroside in vivo by over-expression of the above UGTs.However,as the aglycone substrate of salidroside,the biosynthetic pathway of tyrosol and its regulation are less well understood.The results of related studies revealed that there are two different possibilities for the tyrosol biosynthetic pathway.One possibility is that tyrosol is produced from a p-coumaric acid precursor,which is derived mainly from phenylalanine.The second possibility is that the precursor of tyrosol might be tyramine,which is synthesized from tyrosine.Our previous work demonstrated that over-expression of the endogenous phenylalanine ammonia-lyase gene（PALrs1） and accumulation of p-coumaric acid did not facilitate tyrosol biosynthesis.In contrast,the data presented in our recent work provide in vitro and in vivo evidence that the tyrosine decarboxylase（RsTyrDC） is most likely to have an important function in the initial reaction of the salidroside biosynthesis pathway in R.Sachalinensis.