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研究生:蘇立榛(蘇伊伶)
研究生(外文):Yi-Ling Su
論文名稱:促進鏈球菌透明質酸醱酵產程生理代謝控制之策略
論文名稱(外文):Enhanced Production of Hyaluronic Acid by Streptococcus Fermentation with Control Strategy of Metabolism and Physiology
指導教授:徐泰浩徐泰浩引用關係
指導教授(外文):Tai-Hao Hsu
學位類別:碩士
校院名稱:大葉大學
系所名稱:生物產業科技學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:73
中文關鍵詞:透明質酸獸疫鏈球菌培養溫度抗氧化劑
外文關鍵詞:Hyaluronic acidStreptococcus zooepidemicusculture temperatureantioxidant
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透明質酸(hyaluronic acid)又名玻尿酸或透明多醣體(hyaluronan),由D-葡萄糖醛酸與N-乙醯胺基葡萄糖相互交替連接而成的直鏈多糖體,廣泛存在於動物的組織細胞間質和某些細菌的莢膜。莢膜有無及其大小與菌株所產生之透明質酸酶活性有密切之關係。研究指出活性氧衍生物與自由基能分解透明質酸,而抗氧化劑可抑制氧化壓力所造成之透明質酸分解。本研究探討獸疫鏈球菌(Streptococcus zooepidemicus)醱酵時不同因子如:溫度、起始pH值、攪拌速率及抗氧化劑之添加對生產游離性透明質酸、莢膜性透明質酸及莢膜大小分析之影響,實驗利用影像系統分析菌體莢膜之有無及大小,並分析各因子莢膜性與游離性透明質酸生成之相關性,結果顯示,游離性透明質酸之生成以37℃產量為1.45 g/L、起始pH 7.5產量1.74 g/L、150 rpm產量1.57 g/L、50 mg/L沒食子酸產量1.58 g/L、250 mg/L抗壞血酸產量1.54 g/L、250 mg/L生育醇產量1.49 g/L培養下最高。莢膜性透明質酸的產量上則是在37℃產量0.08 g/L、起始pH 7.5產量0.08 g/L,150 rpm產量0.09 g/L時最好。此外,莢膜大小均在培養6小時最大,此外則隨著培養時間之增加而有減少之情況,而溶血環的作用則與此結果相反。
Hyaluronic acid is a linear polysaccharide composed of repeating subunits ofβ-1,4-linked disaccharide of glucuronic acid β-1,3- N-acetylglucosamine. It has been existed in extracellular matrix in animal tissue. There are close relations in capsule and hyaluronidas .The studies have demonstrated the role played by reactive oxygen-dericed species in depolymerization of hyaluronic acid (HA) , that antioxidants would protect HA from oxidative degradation. The effect of various cultures on temperature, pH, agitation rate and added various antioxidants concentration of hyaluronic acid production by Streptococcus zooepidemicus was studies. And the effects of extracellur hyaluronic acid and capsule hyaluronic acid were investigated. Morphological details of the sample were evaluated using and image analyzer. The result exhibited that extracellur hyaluronic acid concentration of 37℃(1.45 g/L)、pH 7.5 (1.74 g/L)、150 rpm(1.57 g/L)、50 mg/kg of gallic acid (1.58 g/L)、250 mg/kg of ascorbic acid (1.54 g/L)、250 mg/kg of tocopherol (1.49 g/L) had the maximum yield. The capsule hyaluronic acid had the maximum yield in 37℃(0.08 g/L)、pH 7.5(0.08 g/L),150 rpm(0.09 g/L). On the other hand, the capsule were significant increased in 6h by culture time.
中文摘要........................................................ iv
英文摘要........................................................ v
誌謝............................................................ vi
目錄.............................................................vii
圖目錄............................................................x

1. 前言.........................................................1
2. 文獻回顧......................................................3
2.1透明質酸之特性.................................................3
2.1.1透明質酸的由來...............................................3
2.1.2透明質酸的分布...............................................3
2.1.3透明質酸的結構...............................................4
2.2透明質酸之製備與純化............................................7
2.2.1利用動物直接萃取..............................................7
2.2.2利用微生物直接萃取............................................7
2.3透明質酸之生化合成結構...........................................7
2.4透明質酸之性質..................................................11
2.4.1透明質酸之黏彈性..............................................11
2.4.2透明質酸之分子量..............................................12
2.4.3透明質酸之降解反應............................................12
2.5影響透明質酸之因子..............................................15
2.5.1溫度影響.....................................................15
2.5.2 pH值影響....................................................15
2.5.3攪拌速率影響.................................................16
2.5.4抗氧化劑影響.................................................16
2.6透明質酸之應用.................................................17
2.6.1透明質酸於醫藥方面之應用......................................17
2.6.2透明質酸於化妝品方面之應用.....................................19
2.6.3透明質酸於保健食品方面之應用....................................20
3. 材料與方法......................................................21
3.1實驗材料........................................................21
3.1.1實驗藥品......................................................21
3.1.2儀器設備......................................................22
3.2培養基..........................................................22
3.2.1種菌培養基.....................................................22
3.2.2基礎培養基.....................................................23
3.3實驗方法.........................................................23
3.3.1試驗菌株.......................................................23
3.3.2種菌培養.......................................................23
3.3.3種菌保存.......................................................23
3.3.4溶血環試驗.....................................................24
3.4不同溫度之搖瓶培養試驗.............................................24
3.5不同起始pH值之搖瓶培養試驗.........................................24
3.6不同攪拌速率之搖瓶培養試驗..........................................25
3.7添加不同抗氧化劑之搖瓶培養試驗......................................25
3.8游離性透明質酸之試驗...............................................26
3.9莢膜性透明質酸之試驗...............................................26
3.10分析方法........................................................26
3.10.1菌體生質量之測定...............................................26
3.10.2透明質酸含量之測定.............................................27
3.10.3菌體莢膜染色之分析.............................................28
4. 結果與討論.......................................................29
4.1菌體生質量與游離性透明質酸產量之比較................................29
4.1.1溫度之影響.....................................................29
4.1.2起始pH值之影響.................................................32
4.1.3不同轉速之影響.................................................36
4.1.4抗氧化劑之影響.................................................39
4.2莢膜性透明質酸產量之比較..........................................47
4.3溶血性試驗......................................................51
4.4莢膜大小之分析..................................................53
5. 結論............................................................67
參考文獻...........................................................68
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