β-环糊精对海南粗榧悬浮细胞三尖杉酯类碱合成的分析

Analysis of the Production of Cephalotaxine from Suspension Cells of Cephalotaxus mannii by β-Cyclodextrins

海南粗榧(Cephalotaxus mannii)是生产抗癌药物三尖杉酯类生物碱的主要原料。本研究利用不同浓度β-环糊精处理海南粗榧悬浮细胞,通过对细胞悬浮培养过程中细胞生长、产物含量以及代谢途径中关键酶活力分析,研究β-环糊精对海南粗榧生长及次级代谢产物三尖杉酯类碱合成的影响。结果显示,β-环糊精显著提高了三尖杉酯类碱含量并对海南粗榧悬浮细胞的生长有促进作用,添加35 mmol/Lβ-环糊精,产物含量最高(2.26 mg/L),是对照(0.54 mg/L)的4.18倍;悬浮细胞生物量为14.79 g/L,是对照组(9.79 g/L)的1.5倍。同时,β-环糊精提高了细胞6-磷酸葡萄糖脱氢酶以及3-脱氧-D-阿拉伯庚酮糖-7-磷酸合酶活力。因此β-环糊精可通过诱导碳通量转移到磷酸戊糖途径从而促进三尖杉酯类碱的合成,同时也为开发高效诱导子,实现海南粗榧悬浮细胞的工业化生产提供方向。

Cephalotaxus mannii is the main raw material for the production of the anticancer drug Cephalotaxine.In this study,the suspension cells of Cephalotaxus mannii were treated with different concentrations ofβ-cyclodextrin.Through the analysis of cell growth,product content and key enzyme activity in the metabolic pathway during cell suspension culture,the effects ofβ-cyclodextrin on the growth of Cephalotaxus mannii and the synthesis of the secondary metabolite cephalotaxines were studied.The results show thatβ-cyclodextrin significantly increased the production of cephalotaxine and promoted the growth of Cephalotaxus mannii suspension cells.While treated with35 mmol/Lβ-cyclodextrins,the cephalotaxine production was the highest(2.26 mg/L),which was 4.18 times that of the control(0.54 mg/L).The suspended cell biomass was 14.79 g/L,1.5 times that of the control group(9.79 g/L).The activity of glucose 6-phosphate dehydrogenase and 3-deoxy-D-heptanose-7-phosphate synthase was also increased byβ-cyclodextrin.Therefore,β-cyclodextrin can induce the transfer of carbon flux to the phosphopentose pathway to promote the synthesis of cephalotaxines,and also provide direction for the development of effective elicitors to realize the industrialized production of Cephalotaxus mannii cell suspension.