臺灣博碩士論文加值系統

地球上之資源是相當有限的。是故，可再生資源(renewable resource)的利用逐漸受到各方重視。藻類是一種用途廣泛之再生資源，其生質含有豐富的蛋白質、維生素及其他物質可供於食品添加、飼料、醫藥、保健食品等產品。藻類除易於培養與生長快速外，具有不與現有耕作地競爭的特色。
本研究即針對NC64A小球藻進行異營培養，目的在於進行高密度培養與藉由病毒感染綠藻產生透明質酸。本研究之實驗結果發現綠藻NC64A在異營條件下，細胞濃度可以達到3×108cell/L，較自營綠藻之細胞濃度提昇55倍。經由病毒感染後可以分泌產生透明質酸12.6mg/L，分子量為1.6×106Da。而以MSG進行培養，可以有效提昇藻細胞三酸甘油脂(TG)含量，可達乾重之11%。此外，本研究發現EDTA 與單醣同時存在對NC64A會有抑制藻細胞生長之現象。未來本研究擬繼續進行的實驗方向有：
(1)藉由fed-batch碳源繼續提高NC64A藻細胞濃度。
(2)以廢棄物進行藻細胞培養，以降低生產成本。
(3)以雙相(biphasic)策略對藻細胞進行培養，增加其TG含量。
(4)研究不同病毒感染件條對於透明質酸產生之影響。
(5)試圖了解EDTA與單醣對NC64A產生生長抑制的現象。

The energy on the Earth is relatively limited. Therefore, in recent years everybody pay attention to the application and development of renewable resource gradually. Microalga that have a wide range of application are renewable resource. The biomass of microalga include abundant protein, vitamin and other materials that can provide to increase in the food, animal feed, medicine, health care food etc. Easier cultivation and faster growth would the advantage of microalga. Besides, microalga had the special features that never compete in farming ground with traditional farm crop.
This research would aim at the heterotrophic cultivation of Chlorella NC64A with virus PBCV-1 infection for hyaluronic acid production. In the result of our experiment, we discovered that NC64A could cultivate in high cell density with heterotrophic conditions and the cell number was 3×108 cells/L. Compare heterotrophic with autotrophic NC64A, the cell concentration of former could reach 55 times of the latter. The high concentration NC64A were infected with virus PBCV-1(m.o.i.: 0.0014) in 24hours. The molecular weight and concentration of hyaluronic acid, collected from Chlorella NC64A, were 1.6×106 Da and 12.6 mg/L-broth respectively. We observed a phenomenon that Chlorella NC64A would been inhibited by an unclear mechanism when EDTA and monosaccharide were exist in the medium at the same time. In addition, the weight percentage of algae cell triglyceride(TG) has risen to 11% of dry cell weight when alga cultured with MSG as nitrogen source of medium.
In the future, this research would intend and continue the directions of proceeding:
(1) Increase the cell concentration of NC64A continuously by using the strategy of fed- batch carbon source.
(2) Culture algae cell with organic wastes to low the production cost.
(3) Rise the TG content of algae cells with the biphasic strategy.
(4) Study the effect upon the production of hyaluronic acid under using different infection conditions.
(5) Try to understand that how EDTA and monosaccharide induce the inhibition of NC64A growth.

摘 要
誌 謝
目 錄
圖表總錄
第一章 研究動機與目的
1-1 研究動機
1-2 實驗架構圖
1-3 研究方向與範圍
第二章 文獻回顧
2-1 藻類簡介
2-2 CHLORELLA NC64A與PBCV-1
2-3 HA(Hyaluronic Acid)簡介
2-4 環境條件對藻類生長情形之探討
2-5 植物油脂簡介與應用
第三章 分析方法與實驗設備
3-1 菌株
3-2 藻細胞濃度測定
3-3 藻細胞葉綠素含量測定
3-4 病毒PBCV-1活性測試(titer)與保存
3-5 透明質酸與病毒回收程序
3-6 透明質酸之分析方法
3-7 EDTA之HPLC分析方法
3-8 TG之分析方法
第四章 綠藻NC64A培養研究
4-1 前言
4-2 不同EDTA添加濃度對綠藻NC64A生長之影響
4-3 不同培養基組成對綠藻NC64A生長與含脂質量影響
4-4 結論
第五章 以病毒感染綠藻NC64A生產透明質酸
5-1 病毒培養
5-2 透明質酸之生產與分析
5-3 結論
第六章 結論與未來展望
第七章 參考文獻

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