- 期刊
Opportunities for Improving the Plant Cell Culture Processes for Secondary Metabolite Production
改進以植物細胞培養生產二次代謝產物程序之契機
摘要
本文說明一些能改進植物細胞培養程序之方法及手法，並說明相關生化反應器之性能。如何改進細胞生長及促進二次代謝產物之生產性是本文的主要焦點。主要討論的對象為以藜豆(Stizolobium hassjoo)細胞培養生產L-DOPA (3,4-dihydroxy-phenylalanine)之生化程序。對於懸浮細胞培養系統，說明誘導良好癒合組織及細胞之關鍵實驗技術。接著，解說流體剪應力對於細胞絮聚體及二次代謝產物生產之影響。誘導劑的施用方法，反應器內氣相組成之控制及實驗室規模反應器攪拌動力之測定等關鍵性技術均以實例說明。 對於轉殖藜豆毛狀根，首以實驗設計法尋求最佳培養基組成，經培養基最佳化之結果獲致毛狀根質量，L-DOPA含量及L-DOPA生產力分別比未作最佳化時改進了1.8-，2.2-及2.8-倍。噴霧滴流式反應器對於毛狀根之繁殖有甚佳之效果，本文以培養皿固態培養基所得之毛狀根所顯示之特性應用於噴霧滴流式反應器之設計及操作；側根之纏結與施加適當之阻力有利毛狀根之茂密繁殖及L-DOPA之生成。培養末期添加氮源及碳源可有效促進根質量及L-DOPA之產量。
關鍵字
無資料並列摘要
This article illustrates some methods and approaches which improve the performances of plant cell culture processes and relevant bioreactors. The focus is placed on the promotion of cell growth and enhancement of secondary metabolite production. Production of L-DOPA (3,4-dihydroxyphenylalanine) by Stizolobium hassjoo is the primary cell culture system encountered in this article. For suspension cell cultures, the key points for inductions of pertinent callus and cell line are described. Hydrodynamic effects on cell aggregate and secondary metabolite production are demonstrated. Strategy for elicitation, control of gaseous composition in a reactor, measurements of power consumption in shake flasks laboratory-scale agitated reactor are exemplified. For transgenic hairy roots, the medium optimization through statistical experimental design is employed to promote the hairy root weight, L-DOPA content in the roots and L-DOPA productivity. The results show that the root weight, L-DOPA content and L-DOPA productivity promoted 1.8-, 2.2- and 2.8 folds of basal B5 medium control runs. Mist trickling reactor (MTR) is promising for proliferating the hairy roots. The features of hairy root growth on a Petri-dish suggested that the bridging of the lateral roots and exertion of an appropriate hindrance to root tips are favorable to the profuse growth of roots and L-DOPA production. The mode of replenishment of nutrients, especially the nitrogen and carbon sources in the late of the cultures is justified.