摘  要：解脂耶罗维亚酵母菌是一种具较高安全性的柠檬酸生产菌株，该菌易于改造的单细胞结构及较高的柠檬酸产率使其成为极具潜力的工业柠檬酸发酵菌株。研究发现该菌能利用各种碳源发酵生产柠檬酸，其中油脂是其发酵柠檬酸的优良原料。地沟油是一种廉价易得但具有很高社会危害性的不可食用油脂，因此本研究提出将地沟油用于解脂耶罗维亚酵母柠檬酸的生产。首先，将利用各种地摊油作为柠檬酸发酵碳源进行比较，发现地沟油具有较高的柠檬酸发酵产率，可达到15.1 g/l，可作为大规模柠檬酸发酵的碳源；然后通过条件优化，提高地沟油法发酵柠檬酸的产率，经过优化后柠檬酸产量提高至18.9 g/l，本实验的研究将为提高地沟油的合理利用，降低柠檬酸的发酵成本提供理论基础。57753

毕业论文关键词：解脂耶罗维亚酵母菌，地沟油，柠檬酸发酵，发酵条件优化

Abstract：Citric acid is an important organic acid which is widely used in industries. Yarrowia lipolytica is yeast with high yield of citric acid, which is considered safe and more tolerant to contamination. For Y. lipolytica, oil is a preferable carbon resource for citric acid prodcing. Illegal cooking oil is a waste in catering services which is concerned in many courtiers. As a renewable low cost raw material, the main constituent of illegal cooking oil is triacylglycerols and could be used for the formation of citric acid. In this study, citric acid production by Y. lipolytica SWJ-1b from illegal cooking oil compare with other oil was investigated, and we found illegal cooking oil is suitable for citric acid producing which could yield 15.1 g/l of citric acid. Then the compositions in the medium for citric acid production by Y. lipolytica SWJ-1b with illegal cooking oil were optimized, and after the optimization, Y. lipolytica SWJ-1b could gave 18.9 g/l of citric acid.

Key words：Yarrowia lipolytica, illegal cooking oil, citric acid producing, Optimization for citric acid fermentation

目  录

1  前言： 5

2  不同来源的油脂为原料发酵产柠檬酸 7

2.1  材料与试剂 7

2.1.1  菌种 7

2.1.2  培养基和试剂 7

2.2 方法 8

2.2.1  种子液的培养 8

2.2.2  发酵产柠檬酸 8

2.2.3  发酵柠檬酸含量的测定 8

2.3  结果与讨论 9

3  地沟油发酵产柠檬酸的培养条件优化 11

3.1  材料与试剂 11

3.1.1  培养基和试剂 11

3.1.2  菌株 12

3.2  方法 12

3.3  结果与讨论 12

参考文献 16

致谢 18

1  前言：

近年来，随着对能源需求的日益增大，与”能源”相关的新兴企业越来越角逐于现代化能源市场的舞台。柠檬酸作为应用于饮料业、食品业、农业、医药业、纺织业等众多领域的能源之一，其工业化的生产开发存在着广阔的拓展空间：我国国内市场柠檬酸人均年消耗量仅约0.1 kg，是发达国家的人均年消耗量的1/20至1/15。柠檬酸的消耗量与需求量之间，我国经济发展速度不断攀升，而国内人民需求却未充分调动，这种矛盾使得柠檬酸规模化生产的市场前景十分广阔。