枇杷細胞震盪培養及生物反應器培養的動力學研究

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Title:	枇杷細胞震盪培養及生物反應器培養的動力學研究 Kinetic study on the cell of Eriobotrya japonica in suspension and bioreactor culture
Authors:	梁光耀;Kuang-Yao Liang
Contributors:	藥學院藥物化學研究所碩士班
Keywords:	枇杷;懸浮培養;生物反應器;動力學研究;Eriobotrya japonica;suspension;bioreactor;kinetic study
Date:	2010
Issue Date:	2010-09-29 14:10:34 (UTC+8)
Abstract:	枇杷細胞可以生產具生理活性的多種三萜酸化合物。本研究主要探討枇杷細胞懸浮培養的過程中，養分與二次代謝產物之間的動力學關係，進而探討兩者之間的交互作用。固體培養生產有用成分的效率低，因此本研究中建立細胞懸浮震盪培養技術，進行養分與二次代謝產物之動力學研究。並且進一步建立生物反應器的培養技術。利用癒傷組織馴化獲得高產率之細胞株，以懸浮震盪培養系統分析培養基的各種巨量元素，探討細胞生長過程各種養分含量以及細胞生長代謝的動力學曲線。將以上震盪懸浮培養所得資料，作為開發生物反應器試驗之基礎。震盪懸浮培養28天中，細胞乾重變化(3 g/L→13.7 g/L)在第20天之後停止增加，有用二次代謝產物於培養20天後開始快速累積。實驗結果顯示，培養液的磷、碳鈣離子濃度、含糖量及導電度與細胞生長代謝有高度的相關性。快速、方便的導電度測量可作為判別細胞生長情形的指標。而在生物反應器的培養試驗中，顯示利用生物反應器培養可以更縮短生長週期(16天)、產出更多細胞乾重(13.2 g/L)與二次代謝產物(190 mg/g cell dry weight)，其養成分與二次代謝產物形成的動力學曲線與震盪培養雷同，可見枇杷細胞的懸浮培養具有可量化之潛在經濟價值。 The aim of this study is to investigate the relation in nutrient consumption and secondary metabolites kinetics on the suspension culture of Eriobotrya cells. On the previous study, we had established the optimal culture state in loquat tissue culture. However, the solid culture has low efficiency and hard to analysis. In the study, we analysis the kinetic factors between cell growth and metabolism of Eriobotrya cell. Also, the 5L bioreactor culture was established. To induce the suspension cells and high productivity cell line, and establish the analysis methods of scale inorganic elements. The cell weight was recorded, the determination of residual sugar, ions in the medium and secondary metabolites contain were carried out. In 28 days cultivation, the cell weight accumulation increased stopped on day 20, while the secondary metabolites started to accumulate rapidly on day 20. According to the result and data previously, the ions concentration, residual sugar and conductivity had a high relativity with cell dry weight and secondary metabolites production. Especially, the conductivity provides a rapid and convenient way to figure out the growth phase of cell cycle in the cultivation system. In the bioreactor experiment, the result showed that the bioreactor culture has a shorter culture period, better cell growth and better secondary metabolites accumulation. Moreover, the results of kinetic studies in shaker culture and bioreactor had high relativity. It pointed out that suspension culture of Eriobotrya cells is possible to scale up and achieve the commerciality.
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