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題名 | 促進植物細胞生物合成與指標成分的製備=Promote Plant Cell Biosynthesis and the Preparation of Index Components |
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作者姓名(中文) | 吳金濱; | 書刊名 | 中醫藥年報 |
卷期 | 3 2014.12[民103.12] |
頁次 | 頁(33)1-(33)34 |
分類號 | 414.33 |
關鍵詞 | 二次代謝產物; 生物反應器; 三萜酸; Tormentic acid; Secondary metabolites; Bioreactor; Triterpene acids; |
語文 | 中文(Chinese) |
中文摘要 | 摘要研究目的:本計畫運用實驗室已經建立的枇杷懸浮細胞培養的基礎上,探討各種前驅誘導物質或二次代謝物促激劑對枇杷細胞生合成三萜酸的影響。一方面提高細胞培養量、另一方面提高二次代謝產物三萜酸的含量,並且放大到 35公升級的生物反應器進行培養。確立傳統藥材的生物技術利用,並提取純化培養細胞的主要有用成分,供作中藥指標成分之用。 研究方法:透過已馴化的枇杷細胞株,利用生物反應器培養來尋找各種調控因子促進植物細胞生長及合成二次代謝產物方法。培養過程中,每兩天進行採樣,將取得的萃出液進行各種檢測;記錄細胞的生長情形與利用 HPLC分析細胞中二次代謝產物的含量。最後分離純化出三萜酸成分。 結果與討論:生物反應器培養中,最理想的生長環境為 (MS)medium,BA 2.5 ppm,NAA 1 ppm3% sucrose及轉速 80 rpm。蔗糖能提高細胞量,且有利於 Tormentic acid (TA) 的生產,;果糖則有利於 Corosolic acid (CA)的生成。離子添加後,培養至第 20天能達到最高乾重,第 26~30天時,可以達到最高的三萜酸類產量。於第 16天添加 Methyl jasmonate 100mM進行老化誘導可以有效增加細胞乾重 15%,總三萜酸含量增加 23%,TA含量增加 55%。添加醋酸可同步提高所有三萜酸含量。傳統型的轉子可得到較多的細胞重量,離心型可以提高二次代謝產物的產率。於第八天後額外添加 3%蔗糖,使細胞乾重高達 22.42 g/L。細胞於 35L的生物反應器中培養仍有良好的平行放大性。細胞乾重介於 11.48 g/L ~ 12.7 g/L之間,而三萜酸產量介於 151.54 mg/g ~ 135.48 mg/g之間。顯示本試驗細胞株可放大培養。 |
英文摘要 | AIM: The aim of this study is development of the loquat cell lines to find the best method of suspension culture to synthesis second metabolites of triterpenoic acids, as a base study of precussor, elicitor to obtain a large scale of the secondary metabolites of bioreactor culture. Furthermore, use 35 liter bioreactor for a large scale of secondary metabolites syntehsis. Thus, to obtain large scale of secondary metabolites in loquat from suspension culture cells, And purify the bioavtive components for authentic sample of Chinese medicine. METHOD: Through the loquat cell have been domesticated. To use the bioreactor to find a variety of regulatory factors to promote cell growth and synthesis of plant secondary metabolites methods. Sampling every two days and used filter paper to filtrate lequid and cells to obtaine data for various detection. Records of cell growth and HPLC analysis of secondary metabolites. Finally, purified triterpene acid components. RESULTS & DISCUSSION: The great growth environment in bioreactor was MS medium with BA 2.5 ppm, NAA 1 ppm, 3% sucrose, 80 rpm speed. Sucrose could improved all production. Especially, conducived to the production of TA, if changed source to fructose, it couled conducived to the growth of CA. After ion added, the study achieved the highest dry weight In day 20 and the highest triterpene acids yield in day26~30. In day 16, added Methyl jasmonate 100mM could increased 15% cell dry weight, 23% triterpene acids and 55% Tormentic acid. On the other hand, Acetic acid added to increase all the triterpene acid content. The traditional type rotor could availabled much more weight of the cells. Centrifugal type was increased the yield of secondary metabolites. In day 8, Added an additional 3% sucrose could let the dry cell weight up to 22.42 g/L. Cells in the 35 L bioreactor was still a great zoom in parallelism, dry cell weight between12.7 g/L ~ 11.48 g/L, triterpene acids yield between 151.54 mg/g ~ 135.48 mg/g. Those data Showed that loquat cell could zoomed for culture. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。