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目的研究L-色氨酸及其衍生物5-甲基-DL-色氨酸(5-MT)对海南粗榧悬浮细胞生长及三尖杉酯类碱合成的影响。方法在细胞培养第15天,向海南粗榧悬浮培养物中添加一定量的L-色氨酸或5-MT,测定细胞生长、产物量以及代谢途径中关键酶的活力。结果 L-色氨酸与5-MT都能促进三尖杉酯类碱的合成。添加5 mg/L 5-MT时,产物量最高(2.944 mg/L);添加10 mg/L L-色氨酸和5 mg/L 5-MT后,三尖杉酯类碱产量分别是对照(1.343 mg/L)的1.947、2.192倍;而悬浮细胞生物量分别比对照(2.406 g/L)降低了9.35%、18.04%。同时,3-脱氧-D-阿拉伯庚酮糖-7-磷酸合成酶活性分别比对照(0.67 U/mg)降低了12.4%、5.57%;苯丙氨酸解氨酶活性分别比对照(78.21 U/mg)增加了37.72%、28.02%;酚类物质积累比对照(0.36mg/mL)增加了19.14%、9.61%。结论 L-色氨酸及其衍生物5-MT能抑制代谢支路,促进代谢流向产物三尖杉酯类碱的方向。
Objective To study the effects of L-tryptophan and its derivative 5-methyl-DL-tryptophan (5-MT) on the growth of Cynanchum atre and the alkaloid synthesis of harrington in Hainan. Methods On the 15th day of cell culture, a certain amount of L-tryptophan or 5-MT was added to the crude culture suspension of Hainan to determine the cell growth, the amount of the product and the activity of key enzymes in the metabolic pathway. Results Both L-tryptophan and 5-MT promoted the synthesis of harringtonine. The highest amount of product (2.944 mg / L) was obtained when 5 mg / L 5-MT was added, and the production of harringtonine was 10 mg / L L- tryptophan and 5 mg / (1.343 mg / L) 1.947,2.192 times higher than that of the control (2.406 g / L), respectively. The biomass of suspension cells decreased by 9.35% and 18.04%, respectively. At the same time, the activity of 3-deoxy-D-arabinoheptaose-7-phosphate synthase was reduced by 12.4% and 5.57%, respectively, compared with the control (0.67 U / mg); the activity of phenylalanine ammonia lyase was 78.21 U / mg) increased by 37.72% and 28.02%, respectively. Phenolic accumulation increased by 19.14% and 9.61% compared with the control (0.36mg / mL). Conclusion L-tryptophan and its derivatives 5-MT can inhibit the metabolism of the branch, and promote the metabolic direction of the product of harringtonine base direction.