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研究生:黃惠琴
研究生(外文):Hui-Chin Huang
論文名稱:樟芝菌絲體深層培養之研究
指導教授:楊芳鏘楊芳鏘引用關係
指導教授(外文):Fan-Chianf Yang
學位類別:碩士
校院名稱:東海大學
系所名稱:化學工程學系
學門:工程學門
學類:化學工程學類
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:103
中文關鍵詞:樟芝環境因子回應曲面法
外文關鍵詞:Antrodia cinnamomeaphysical and chemical factorsRSM
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本研究以Antrodia cinnamomea CCRC 35396為主要試驗菌株,利用三角瓶液態培養,探討不同物理及化學環境因子,對樟芝菌絲體生長之影響,希望藉由培養因子的調整與控制提高菌絲生長速率及菌體濃度。
主要實驗項目:以三角瓶批式培養,探討時間、pH、溫度、轉速、表面通氣、培養成分等因子對菌絲生長與多醣生成之影響,並利用回應曲面法提高菌絲體產量。
結果顯示:在250ml三角瓶批式培養中,樟芝菌絲生長的最適培養溫度為25℃,初始pH值為5,轉速為100rpm,在此條件下培養14天菌體濃度達486.8mg/100ml。過高的表面通氣效應及過快的振盪轉速皆不利於菌絲球生長,最適迴轉速率在100rpm左右。在碳、氮源種類方面:玉米粉及YM Broth對菌體生長有較佳表現。
利用回應曲面法(RSM)探討碳源濃度、氮源濃度和初始pH值因子交錯影響,決定最佳培養基組成的條件下,培養7天菌體濃度可達到2164.2mg/100ml,產量比原濃度提昇5倍以上。本研究結果證實利用三角瓶培養條件控制,配合回應曲面法培養組成決定,可以有效的縮短樟芝培養時間,並且能夠快速提高菌絲體產量。
In this study the influence of different physical and chemical factors on the production of biomass and polysaccharide by Antrodia cinnamomea CCRC 35396 was investigated in the submerged culture of using shake flasks. Optimal cultural conditions were expected to decide to enhance the growth rate and the mycelium concentration.
The main concerned cultural conditions include: time, pH, temperature, rotation speed, surface aeration and medium compositions, etc. In additions, Response Surface Methodology (RSM) was used to optimize the medium composition in order to increase the mycelium concentration.
The results indicated that when the submerged culture in shake flasks was operated at 25℃, initial pH 5 and rotation speed 100rpm, the mycelium concentration reached to 486.8mg/100ml at 14th day. Both the high surface aeration rate and rotation speed were unfavorable to the growth of mycelium. The optimal rotation speed was shown to be around 100rpm. After the comparison of various sources of carbon and nitrogen, corn powder and YM broth were selected for the further tests of RSM.
According to the RSM, the medium composition was modified and the mycelium concentration of Antrodia cinnamomea reached to 2164.2mg/100ml at the 7th day and had a 5-fold increase, compared to the growth in basal medium. The results proved that the production rate of mycelium of Antrodia cinnamomea could be highly enhanced by means of the control of cultural conditions and the modification of medium compositions based on the RSM.
目錄
頁次
中文摘要…………………………………………Ⅲ
英文摘要…………………………………………IV
目錄……………………………………………… V
表目錄…………………………………………VIII
圖目錄……………………………………………X
照片目錄…………………………………………XII
第一章 緒論…………………………………1
1-1 前言……………………………………………1
1-2 高等擔子菌菇類的介紹…………………2
1-3 菇類菌絲體深層培養在食品工業上之應
用………………………………………………3
1-4 本文大綱……………………………………5
第二章 文獻回顧…………………………6
2-1 樟芝的介紹…………………………………6
2-1-1 樟芝的分類……………………………6
2-1-2 樟芝的成分分析………………………8
2-2 高等蕈類深層培養與生長型態………13
2-2-1菌絲球的形成與構造………………15
2-2-2深層培養的環境影響因素………17
2-3 菇類多醣的介紹…………………………21
2-3-1多醣回收………………………………23
2-4 回應曲面法的介紹………………………26
2-4-1 設計原理……………………………26
2-4-2 二水準因子設計…………………27
2-4-3 陡升路徑法…………………………30
2-4-4 中心混成設計………………………32
2-4-5 回應曲面模式適切性之統計檢驗33
第三章 實驗材料與分析方法…………34
3-1 菌株…………………………………………34
3-2 實驗藥品……………………………………34
3-3 實驗儀器與設備…………………………35
3-4 組成分析方法……………………………36
3-4-1 pH值測定…………………………36
3-4-2 固態培養菌絲生長測定…………36
3-4-3 菌體粒徑測定……………………36
3-4-4 菌絲球密度測定…………………36
3-4-5 菌體濃度測定……………………37
3-4-6 葡萄糖濃度測定…………………37
3-4-7 多醣濃度測定………………………37
3-5 攪拌式反應器……………………………39
第四章 實驗方法……………………………40
4-1 斜面培養……………………………………40
4-1-1 試管斜面培養基製備與接菌活化40
4-1-2 培養皿平面培養基製備與接菌活
化………………………………………40
4-2 液態基礎培養基組成……………………41
4-3 種菌與接菌量……………………………42
4-4 菌種篩選……………………………………43
4-5 三角瓶培養試驗─物理因素……………44
4-6 三角瓶培養試驗─化學因素……………46
4-7 三角瓶培養試驗─回應曲面法(RSM)49
4-8 發酵槽培養試驗…………………………52
第五章 結果與討論…………………………53
5-1 菌種篩選……………………………………53
5-2 三角瓶培養試驗─物理因素……………57
5-3 三角瓶培養試驗─化學因素……………68
5-4 三角瓶培養試驗─回應曲面法(RSM)74
5-5 發酵槽培養試驗…………………………87
第六章 結論與展望…………………………90
6-1 結論…………………………………………90
6-2 未來展望……………………………………91
參考文獻………………………………………95
附錄……………………………………………101
簡歷……………………………………………103
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