博碩士論文 102324037 詳細資訊




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姓名 蕭雯如(Wen-ju Hsiao)  查詢紙本館藏   畢業系所 化學工程與材料工程學系
論文名稱 探討靈芝在液態發酵中與酵母菌共培養並利用兩階段式操作以增加靈芝酸產量
(Enhancement of ganoderic acids production of Ganoderma lucidum by co-culture with Saccharomyces cerevisiae.)
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摘要(中) 靈芝菌為食藥用菇中的一種,食藥用菇最吸引人的地方在其抗癌、抗腫瘤的效果,而靈芝含有特殊三萜類(特稱為「靈芝酸」),為其他植物所沒有,靈芝中所含三萜類的主要功能為:抑制癌細胞生長、抑制組織胺釋放,防止過敏、促進肝功能、促進血小板凝集、以及降血脂,防中老年人發生心血管方面的疾病,因此具有相當潛力被開發為未來醫療用品,或作為近年來國內相當熱門的保健食品。

靈芝主要生長在腐樹或樹木的根部,屬於非褶菌目,靈芝科。由於其子實體生長緩慢,野生靈芝很少見到且質量不容易控制,導致在市面上的價格相當昂貴,難以廣泛應用於醫療用藥品。因此當今以人工培養的方式來進行學術上研究以及提供市場上需求,其中以液態培養具有相當潛力,其優點為:菌絲體生長速度較快、所需空間較小、汙染的機率較低,基於上述幾點描述,此研究將利用液態培養的方式來增加總三萜的含量。

過去增加總三萜的方法,大部分著重於培養基的改變,以及環境因子的操控,近年有學者探討兩階段式發酵,由於三萜的生產是因為靈芝菌受到環境限制而產生的二次代謝物,因此第一階段讓菌體快速生長,第二階段在菌體生長到最高峰時改變環境因子增加總三萜產量。

本計畫嘗試開發兩階段發酵製程系統,在第二階段加入酵母菌進行共培養來提升總三萜的產量,並探討共培養機制。實驗結果顯示,第二階段以靜置操作及酵母菌接菌量5%(v/v)能產出最高三萜量171.5 mg/g DW,由實驗結果可發現,酵母菌所產生的酒精為三萜產量增加之主因。

摘要(英) Ganoderma lucidum is one of the medicinal mushrooms. The most attractive characteristic of the medicinal mushrooms is its anti-tumor effect. Ganoderma lucidum has gained wide popularity as a health food and became the most valuable mushroom in Taiwan.

However, because of its host specificity, slow growth rate and rarity in nature, the fruiting bodies of Ganoderma lucidum have become expensive mushrooms in recent years. Thus, investigators have exerted their efforts to prepare this mushroom from submerged culture.

The main objectives of this proposal are to develop two-stage fermentation system. In first stage, we make the Ganoderma lucidum grow in high speed for 8 days, and Saccharomyces cerevisiae was added to the fermentation broth in second stage with aerated or static process to enhance ganoderic acids production. The amount of ganoderic acids reached 171.5 mg/g DW by using static operation with 5%(v/v) of inoculation level of the Saccharomyces cerevisiae in 250 ml shake flask.

A novel two stage operation to optimize triterpenoids production of a co-culture was proposed and successfully demonstrated, reaching highest ganoderic acids content obtained in this study.

關鍵字(中) ★ 靈芝
★ 靈芝酸
★ 共培養
★ 酵母菌
關鍵字(英) ★ Ganoderma lucidum
★ ganoderic acids
★ co-culture
★ Saccharomyces cerevisiae
論文目次 目錄

中文摘要………………………………………………………….………...……i

英文摘要…………………………………………………………………….….iii

致謝……………………………………………………………………………..iv

目錄……………………………………………………………..………….……v

表目錄……………………………………………………………..................... ix

圖目錄……………………………………………………………...…................x

第一章、 緒論…………………………………………….…………….……….1

1-1 研究動機…………………………………………………………….....1

1-2 研究目的……………………………………………………….............2

第二章、 文獻回顧……………………………………………………………...3

2-1 菇類介紹……………………………………………………………….3

2-2 靈芝介紹……………………………………………………………….4

2-3 靈芝的成分分析……………………………………………...………..5

2-4 靈芝之藥理效用文獻考察………………………………...…………..6

2-4-1 降血壓作用…………………………………………………...…6

2-4-2 抗腫瘤作用…………………………………………………...…6

2-4-3 抗病毒作用…………………………………………………...…6

2-4-4 保肝作用……………………………………………………...…6

2-4-5 降血糖作用……………………………………………………...7

2-4-6 免疫調節………………………………………………………...7

2-4-7 抗氧化作用……………………………………………………...7

2-5 真菌多醣介紹………………………………………………...………8

2-5-1真菌多醣的特性…………………………………………………8

2-5-2真菌多醣的結構………………………………………………..12

2-6 真菌三萜類的介紹………………………………………………….15

2-6-1 萜類的重要性質……………………………………………….15

2-6-2 合成萜類的單體………………………………………...……..15

2-6-3 三萜的合成機制……………………………………………….16

2-6-4 菇類三萜的基本結構………………………………………….19

2-7 液態發酵培養…………………………………….…………………..23

2-7-1 何謂液態發酵培養…………………………………………….23

2-7-2 液態發酵培養的優點………………………………………….23

2-8 影響發酵的物理與化學因子……………..………………………….24

2-8-1 物理因子……………………………………………………….24

2-8-2 化學因子……………………………………………………….27

2-9 共培養……………………………………………………………...…30

2-9-1 Rhodotorula glutinis 與 Spirulina platensis 共培養…………30

2-9-2 Chlorella sp. 與 Saccharomyces cerevisiae 共培養………….31

2-9-3 Trichoderma reesei 與 Aspergillus niger 共培養………….…31

2-9-4 Saccharomyces cerevisiae 與Pichia stipites共培養……….…32

第三章、 材料與方法………………………………………………………….33

3-1 實驗材料……………………………………………………………...33

3-1-1 實驗菌株………………………………………………...……..33

3-1-2 實驗藥品……………………………………………………….34

3-1-3 實驗儀器及其他設備………………………………………….35

3-2 實驗方法……………………………………………………...………36

3-2-1 菌株保存……………………………………………………….36

3-2-2 培養基組成…………………………………………………….36

3-2-3 種菌操作條件………………………………………………….39

3-2-4 單培養實驗…………………………………………………….39

3-2-5 共培養實驗…………………………………………………….40

3-2-6 Saccharomyces cerevisiae 共培養之接菌量……………….….41

3-2-7 共培養機制探討……………………………………………….41

3-2-8 Scaling-up 2L發酵槽………………………..…………………41

3-4 分析方法…………………………………………………………...…42

3-4-1 發酵液處理流程…………………………………………….…42

3-4-2 菌重分析……………………………………………………….43

3-4-3 葡萄糖殘量分析(還原醣測定)………………………….…….43

3-4-4 酸鹼值測定(pH值)……………………………………………44

3-4-5 總三萜分析…………………………………….………………44

第四章、 結果與討論………………………………………….………………46

4-1 Ganoderma lucidum 與Saccharomyces cerevisiae單培養之結果….46

4-2 兩階段式操作…………………………………………………….…47

4-2-1 兩階段式共培養對菌重的影響……………………………….47

4-2-2兩階段式共培養對靈芝酸的影響……………………………..48

4-3 酵母菌接菌量對產物的影響……………………………………….49

4-4 共培養機制探討…………………………………………….………50

第五章、 結論………………………………………………………………….52

第六章、 參考文獻…………………………………………………………….53

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指導教授 徐敬衡(Chin-hang Shu) 審核日期 2015-8-27
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