Liu Zhi, Yu Long-Jiang, Li Chun-Yan, Zhao Chun-Fang
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2005 Apr;31(2):199-204.
There is a dichotomy in the biosynthetic pathway of terpenoid precursor isopentenyl diphosphate (IPP) in higher plant. One is the classical mevalonate pathway in cytosol, and the other is non-mevalonate pathway in plastid. To know the origin of the taxane ring system of taxol in suspension culture of Taxus chinensis, lovastatin and fosmidomycin were used to block the 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) and 1-deoxy-D-xylulose-5-phosphate reducto-isomerase (DXR) in the mevalonate and non-mevalonate branch respectively of the terpenoid biosynthetic pathway. Methyl jasmonate (MJ) was used to improve the biosynthesis of taxol. Taxol content was determined by HPLC, the transcriptional expression of genes encoding DXR and HMGR were investigated by real time PCR. Taxol production was lowered by about 2/5 and 1/5 by fosmidomycin (200 mmol/L) and fosmidomycin (200 mmol/L)+MJ (100 mmol/L) treatment respectively, and was lowered by about 1/6 and 1/10 by lovastatin (1 mmol/L) and lovastatin (1 mmol/L) + MJ (100 mmol/L) respectively, which means that both mevalonate and non-mevalonate pathway contribute to taxol biosynthesis, and the latter is the main source of IPP. Inhibitors lovastatin and fosmidomycin both promoted the transcriptional expression of hmgr and dxr, which indicated a metabolic cross talk between cytosolic and plastidial pathways of taxol biosynthesis.
高等植物中萜类前体异戊烯基二磷酸(IPP)的生物合成途径存在二分法。一种是胞质溶胶中的经典甲羟戊酸途径,另一种是质体中的非甲羟戊酸途径。为了了解中国红豆杉悬浮培养物中紫杉醇紫杉烷环系统的起源,使用洛伐他汀和磷霉素分别阻断萜类生物合成途径中甲羟戊酸和非甲羟戊酸分支中的3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)和1-脱氧-D-木酮糖-5-磷酸还原异构酶(DXR)。茉莉酸甲酯(MJ)用于改善紫杉醇的生物合成。通过高效液相色谱法测定紫杉醇含量,通过实时PCR研究编码DXR和HMGR的基因的转录表达。磷霉素(200 mmol/L)和磷霉素(200 mmol/L)+MJ(100 mmol/L)处理分别使紫杉醇产量降低约2/5和1/5,洛伐他汀(1 mmol/L)和洛伐他汀(1 mmol/L)+MJ(100 mmol/L)分别使紫杉醇产量降低约1/6和1/10,这意味着甲羟戊酸途径和非甲羟戊酸途径都对紫杉醇生物合成有贡献,且后者是IPP的主要来源。抑制剂洛伐他汀和磷霉素均促进了hmgr和dxr的转录表达,这表明紫杉醇生物合成的胞质溶胶途径和质体途径之间存在代谢相互作用。