臺灣博碩士論文加值系統

本研究主要探討添加溶菌酶及過氧化氫對Streptococcus zooepidemicus ATCC 39920生產玻尿酸之影響。雖然此二添加物對菌體具傷害性，但研究發現，在醱酵過程對數生長初期適量添加過氧化氫及誘導期適量添加溶菌酶，將有助於玻尿酸產量之提升。

The effect of lysozyme and hydrogen peroxide on the Streptococcus zooepidemicus ATCC 39920 which produce hyaluronic acid was investigated. These two additional agents were harmful to streptococcal cells, but improved the production of hyaluronic acid by adding appropriate hydrogen peroxide in logarithmic growth phase and appropriate lysozyme in lag phase.

總目錄
中文摘要..................................................I
英文摘要.................................................II
誌謝....................................................III
總目錄...................................................IV
表目錄..................................................VII
圖目錄.................................................VIII
第一章 緒論...............................................1
1-1 前言..................................................1
1-2 玻尿酸的來源..........................................2
1-3 玻尿酸的結構..........................................3
1-3-1 一級結構............................................3
1-3-2 二級結構............................................3
1-3-3 三級結構............................................3
1-4 玻尿酸生化合成途徑....................................6
1-5 玻尿酸之物理化學性質..................................8
1-5-1 黏彈性..............................................8
1-5-2 降解反應............................................8
1-6 玻尿酸的應用..........................................9
1-7 醱酵策略.............................................10
1-8 研究動機.............................................12
1-8-1 溶菌酶刺激鏈球菌生產玻尿酸原理.....................13
1-8-2 過氧化氫刺激鏈球菌生產玻尿酸原理...................14
第二章 實驗材料與方法....................................17
2-1 實驗菌株及藥品.......................................17
2-1-1 實驗菌株...........................................17
2-1-2 實驗藥品...........................................17
2-2 實驗儀器與實驗裝置...................................18
2-2-1 實驗儀器...........................................18
2-2-2 實驗裝置...........................................20
2-3 培養基的組成.........................................20
2-3-1 前培養.............................................20
2-3-2 批次培養...........................................20
2-4 實驗方法.............................................23
2-4-1 菌株保存...........................................23
2-4-2 菌株活化...........................................24
2-4-3 前培養.............................................24
2-4-4 主醱酵實驗.........................................24
2-5 批次醱酵實驗流程.....................................24
2-6 分析方法.............................................25
2-6-1 菌體濃度分析方法...................................25
2-6-2 玻尿酸濃度分析方法.................................28
2-6-3 醱酵液黏度分析.....................................31
第三章 結果與討論........................................33
3-1 添加溶菌酶之醱酵策略.................................33
3-1-1 添加劑量範圍.......................................33
3-1-2 最佳添加量.........................................36
3-1-3 最佳添加時機.......................................39
3-2 添加過氧化氫之醱酵策略...............................47
3-2-1 添加劑量範圍及時間.................................49
3-2-2 最佳添加量.........................................52
第四章 結論..............................................56
第五章 參考文獻..........................................57

表目錄
表2-1 TSB 前培養基組成...................................21
表2-2 批次醱酵培養基組成.................................21
表3-1 搖瓶實驗添加溶菌酶量與玻尿酸濃度間關係.............35
表3-2 添加不同溶菌酶量下菌體量及玻尿酸產量之結果表格.....38
表3-3 於第一小時添加不同溶菌酶量下菌體及玻尿酸產量之表格.42
表3-4 於第三小時添加不同溶菌酶量下菌體及玻尿酸產量之表格.44
表3-5 於第五小時添加不同溶菌酶量下菌體及玻尿酸產量之表格.46
表3-6添加過氧化氫玻尿酸溶液動黏度變化情形...............48
表3-7 添加過氧化氫醱酵液動黏度變化情形...................51
表3-8 添加不同量過氧化氫下菌體量及玻尿酸產量之結果表格...54

圖目錄
圖1-1 玻尿酸的雙醣單位(一級結構)..........................4
圖1-2 玻尿酸的二級結構....................................4
圗1-3 玻尿酸分子在溶液中的網狀結構........................5
圖1-4 玻尿酸生化合成示意圖................................7
圖1-5 玻尿酸生化合成途徑..................................7
圖1-6 鏈球菌於醱酵過程其莢膜的變化.......................15
圖1-7 鏈球菌結構.........................................15
圖1-8 過氧化氫降解玻尿酸關係圖...........................16
圖1-9 碳源於鏈球菌代謝過程中利用方向示意圖...............16
圖 2-1 批次醱酵裝置圖....................................22
圖 2-2 批次醱酵實驗流程圖................................26
圖 2-3 葡萄糖醛酸濃度檢量線..............................30
圖 2-8 Ubbelodhe 黏度計..................................32
圖3-1 添加不同溶菌酶量下菌體量及玻尿酸產量之曲線圖.......37
圖3-2 第一小時添加不同溶菌酶量菌體量及玻尿酸產量曲線圖...41
圖3-3 第三小時添加不同溶菌酶量菌體量及玻尿酸產量曲線圖...43
圖3-4 第五小時添加不同溶菌酶量菌體量及玻尿酸產量曲線圖...45
圖3-5 相對溶氧值曲線.....................................51
圖3-6 添加不同過氧化氫下菌體量及玻尿酸產量之曲線圖.......53

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