臺灣博碩士論文加值系統

利用攪拌式生物反應槽培養顆粒狀的鬚根黴，以葡萄糖為碳源，開發一種提高L型乳酸產量的醱酵方法。醱酵過程不用碳酸鈣，而是用氨水控制酸鹼度，由於氨水對於黴菌細胞有毒性，造成乳酸產量降低。因此於氨水中添加碳酸氫銨或於培養基中添加尿素，可降低氨水對黴菌細胞所造成的傷害，提高乳酸的產量。醱酵控制於pH 5.0以及每升1克的尿素加量，中和劑是10% (w/v)氨水添加2% (w/v)的碳酸氫銨，醱酵進行48小時後，得到L型乳酸的最終濃度是每升98克，葡萄糖轉換成乳酸的轉化率是82%。重複批次醱酵，最初的四槽得到L型乳酸的最終濃度逐漸降低，依次為：每升98、95、88以及82克；完成醱酵的時間依次為：48、48、52以及54小時。

Pellet-form Rhizopus arrhizus was cultured in a stirred tank bioreactor for enhanced L-lactic acid production using glucose as substrate and an ammoniacal solution as neutralizing agent. Ammonium hydroxide may cause damage to fungal cells in terms of “chemical burn”, leading to a poor production of lactic acid. It was found when ammonium bicarbonate was added to the ammonia water or urea was added to the culture medium, the toxicity of ammonium hydroxide decreased and lactic acid production was enhanced. The fermentation was carried out at pH 5.0 with the addition of 1 g L–1 urea in the culture medium, using an ammoniacal solution as neutralizing agent, which was 2% (w/v) ammonium bicarbonate in 10% (w/v) ammonia water. The batch fermentation completed in 48 h. The final concentration of lactic acid was 98 g L–1 with a yield of 82%. However, the final concentrations of lactic acid obtained in the first four cycles of repeated batch fermentation were 98, 95, 88 and 82 g L–1, with fermentation times being 48, 48, 52 and 54 h, respectively.

ACKNOWLEDGEMENTS...........................................I
ABSTRACT...................................................................II
中文摘要...................................................................…III
List of Figure...............................................................VII
List of Table ............................................................…..X
CHAPTER 1. INTRODUCTION.................................1
1.1. Lactic acid…………………………………………………1
1.1.1. Properties of lactic acid…………………………………………1
1.1.2. Manufacturing technologies…………………………………….2
1.1.2.1. Chemical synthesis……………………………………..3
1.1.2.2. Carbohydrate fermentation……………………………..4
1.1.3. Current uses……………………………………………………..7
1.1.4. Potential productions and markets………………………………8
1.2. L-lactic acid………………………………….…………...11
1.3. Poly-L-lactic acid ( PLLA )……………………………..11
1.4. Rhizopus oryzae………………………………………….16
1.5. Aim of the study...……………………………………….20
CHAPTER 2. MATERIALS AND METHODS........22
2.1. Materials…………………………………………………22
2.1.1. Microorganisms………………………………………………..22
2.1.2. Instruments…………………………………………………….22
2.1.3. Chemicals……………………………………………………...23
2.2. Culture of R. arrhizus…………………………………...25
2.2.1. Culture medium………………………………………………..25
2.2.2. Culture method………………………………………………...26
2.3. Analytical methods……………………………………...32
2.3.1. Determination of biomass…….………………………………..32
2.3.2. Analysis of lactic acid and glucose………………………….....32
2.4. Monitoring and Control of fermentation……………...35
CHAPTER 3. RESULTS AND DISCUSSIONS........39
3.1. Mycelial morphology of R. arrhizus in submerged culture………………………………………………….……..39
3.2. Lactic acid fermentation using ammonia water as neutralizing agent………..…………………………..………44
3.3. Repeated batch fermentation .……………….………....50
CHAPTER 4. CONCLUSION....................................52
REFERENCE...............................................................53
APPENDIXS…………………………………….........61
Appendix 1. Conventional method of production of L-lactic acid….………………………………………………….……..61
Appendix 1. Conventional method of production of L-lactic acid….………………………………………………….……..62

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