烟酸、天冬氨酸和异亮氨酸对雷公藤悬浮细胞生长及次生代谢产物含量的影响

摘要

为明确烟酸、天冬氨酸和异亮氨酸对雷公藤次生代谢产物含量的影响,通过添加烟酸、天冬氨酸和异亮氨酸来分析雷公藤(Tripterygium wilfordii Hook.f.)悬浮细胞中雷公藤内酯醇、雷公藤吉碱和雷公藤次碱产物的含量.采用HPLC检测分析烟酸、天冬氨酸和异亮氨酸3种物质添加后雷公藤茎悬浮细胞中雷公藤内酯醇、雷公藤吉碱、雷公藤次碱的含量,结果表明,低浓度(0.5 mmol/L)烟酸处理能显著促进细胞的生长,其生长速率为对照的1.15倍,但对细胞干重增加影响不大;不同浓度的异亮氨酸处理能极大促进细胞中雷公藤内酯醇、雷公藤吉碱和雷公藤次碱的产生,并分别在0.1、1.5、0.1 mmol/L浓度下达到最大值,为4.65、30.24、166.41μg/g,但培养液中次级代谢产物的分泌则相对减少;异亮氨酸对总的雷公藤吉碱和雷公藤次碱含量的增加有促进作用;天冬氨酸处理则主要促进细胞中雷公藤吉碱的产生和培养液中次碱的分泌,并在0.5 mmol/L浓度下其含量达到110.09μg/g、4.06μg/瓶;烟酸处理则相对抑制次级代谢产物的产生和分泌.异亮氨酸处理悬浮细胞可以很大程度上增加雷公藤吉碱,雷公藤次碱的产量,推测异亮氨酸可能参与了雷公藤生物碱的生物合成,该结果的应用对雷公藤细胞培育生产次级代谢产物有重要意义.%In order to investigate the effects of different compounds on cell growth and the production of secondary metabolites in Tripterygium wilfordii Hook.f.suspension cell culture,the different compounds of niacin,aspartic acid and isolencine were analyzed in the hope of improving the content of terpenoid and alkaloid in T.wilfordii,which might provide the further information to enhance the content of T.wil-fordii suspension cell.The content of triptolide,wilforgine and wilforine was analyzed by HPLC in differ-ent samples of T wilfordii suspension cell.The niacin could prominently promote the growth of suspen-sion cell in low concentration of 0.5 mmol/L,the rate of growth was 1.15 times that of the control,butit had little effect on the increase of cell dry weight.The isoleucine (Ile)increased the content of triptolide, wilforgine and wilforine in different concentrations,the contents of triptolide,wilforgine and wilforine were respectively 4.65,30.24 and 166.41μg/g dry weight when the concentration of isoleucine was 0.1,1.5and 0.1 mmol/L,but the secretion of triptolide,wilforgine and wilforine was low in the culture medium.The precursors of isolencine,aspartic acid and niacin had different roles in regulating the biosynthesis of triptol-ide,wilforgine and wilforine.The isoleucine promoted the production of total of wilforgine and wilforine in culture cell and culture medium,the aspartic acid could increase the production of wilforgine and the secre-tion of wilforine into medium.The total of wilforgine and wilforine was 110.09 μg/g dry weight and 4.06μg/flaskin 0.5 mmol/L of isoleucine and aspartic acid.The niacin inhibited the production of triptol-ide,wilforgine and wilforine.The isoleucine promoted the production of total of wilforgine and wilforine in culture cell and culture medium in T.wilfordii cell suspension culture.It was presumed that isoleucine might be involved in the sesquiterpene alkaloids synthesis.The results have great significance in suspension cell culture and the production of secondary metabolites in T.wilfordii.