掌葉大黃毛狀根之建立及其藥用蒽醌成份之生產條件檢討

The Establishment Of Rheum palmatum L. Hairy Root Culture And The Production Of Anthraquinone By The Culture Thereof

林怡辰(Yi-Chen Lin)

指導教授 ： 陳建源

國立臺灣大學/生命科學院/微生物與生化學研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.10243

摘要

利用農桿根群菌(Agrobacterium rhizogenes) A4 野生型菌株感染掌葉大黃外植體誘導毛狀根以建立掌葉大黃毛狀根的培養系統，再經由PCR方法進行確認。檢討該毛狀根之最適培養基組成，並且檢討擴大培養之最適培養條件。分別利用比色法、薄層層析法和高效能液相層析法分析毛狀根中游離蒽醌化合物的含量。
本研究以大黃酸作為標準品，在搖瓶培養中測試其最適培養基組成。由實驗結果得知，以B5培養基含2%蔗糖、20.25 mM硝酸鉀、6.75 mM硫酸銨和0.75 mM磷酸二氫鈉為最適培養基組成分。利用最適化之B5培養基進行掌葉大黃毛狀根的培養，培養42天後可得到最高大黃酸產量10.06 mg/L，與搖瓶培養所得之7.264 ± 0.06 mg/L相比較，大黃酸產量增加了1.38倍。擴大培養係利用1公升之氣泡塔式生物反應器(Bubble column reactor)，以B5培養基為基礎培養基培養49 天，可得到的大黃酸產量為17.245 mg/L，與搖瓶培養所得之7.264 ± 0.06 mg/L相比較，大黃酸產量增加了2.37倍。

關鍵字

大黃酸 ； 毛狀根 ； 掌葉大黃

並列摘要

Culture of Rheum palmatum L. hairy roots were established after transformation with Agrobacterium rhizogenese A4, and the transformation was confirmed by PCR for the presence of a T-DNA sequence in their genomes. Quantitative determination of anthraquinones were performed by colorimetry, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). Among these anthraquinones produced by Rheum palmatum L., emodin and rhein were considered more important. In the studies, rhein is taken as the reference standard. Root growth and production of rhein were investigated in various basal media. It was found that B5 medium was superior to MS medium in terms of both root growth and rhein production. The results indicate that Rheum palmatum L. hairy root can be cultivated in the B5 medium enriched with 2 % sucrose, 20.25 mM KNO3, 6.75 mM NH4NO3 and 0.75 mM NaH2PO4. The rhein production is 10.06 mg/L in a 42 days` culture. The productivity is 1.38 folds as that in normal flask culture. A scale up culture of Rheum palmatum L. hairy root from 125 mL flask to 1 L bubble column reactor was conducted. The rhein production is 17.245 mg/L in a 49 days` culture. The productivity is 2.37 folds as that in flask culture.

並列關鍵字

Rhein ； Rheum palmatum L. ； Hairy Root

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