CN111560316A - 一种腺嘌呤生产菌种用培养基 - Google Patents
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Abstract
本发明公开了一种腺嘌呤生产菌种用培养基，它包含如下含量的组分：葡萄糖10‑100g/L，酵母粉5‑15g/L，硫酸铵2‑10g/L，磷酸二氢钾0.5‑2.5g/L，磷酸氢二钾1‑2.5g/L，其他氮源5‑25g/L，玉米浆2‑10g/L，微量元素0.2‑1g/L。原材料来源广泛、成本低廉，由它培养的菌种质量稳定、菌种发酵单位高。
Description
技术领域
本发明属于菌种培养基技术领域，尤其是一种腺嘌呤生产菌种用培养基。
背景技术
腺嘌呤，是DNA和RNA的重要组成部分，是众多医药原料的关键中间体，主要用于生产抗艾滋病药物替诺福韦酯、抗乙肝病毒药物阿德福韦酯等。
目前市场的腺嘌呤产品大部分采用有机合成的方法，生产过程中使用大量的、多种类有机溶剂，存在着生产制造成本高、安全隐患大的问题。部分厂家使用微生物发酵法生产腺嘌呤，但是由于发酵单位偏低导致制造成本相对有机合成没有优势，市场竞争力不大。
发明内容
本发明的目的在于克服现有技术的缺陷，提供一种腺嘌呤生产菌种用培养基，原材料来源广泛、成本低廉，由它培养的菌种质量稳定、菌种发酵单位高。
本发明的技术方案是这样实现的：一种腺嘌呤生产菌种用培养基，它包含如下含量的组分：葡萄糖10-100g/L，酵母粉5-15g/L，硫酸铵2-10g/L，磷酸二氢钾0.5-2.5g/L，磷酸氢二钾1-2.5g/L，其他氮源5-25g/L，玉米浆2-10g/L，微量元素0.2-1g/L。
进一步，上述培养基配方中，酵母粉优选啤酒酵母浸粉和面包酵母浸粉。
进一步，上述培养基配方中，其他氮源包括大豆蛋白胨、豆粕粉、黄豆粉、玉米蛋白、大米蛋白中的一种或者多种。
进一步，上述培养基配方中，微量元素是指生物素、鸟嘌呤、次黄嘌呤、维生素B12、维生素B1中的多种。
钾1g/L，玉米蛋白5g/L，大米蛋白5g/L，玉米浆10g/L，鸟嘌呤0.1g/L，次黄嘌呤0.1g/L，维生素B1 0.1g/L，维生素B12 0.1g/L。
实施例4：一种腺嘌呤生产菌种用培养基，它包含如下成分的组分：葡萄糖20g/L，啤酒酵母浸粉10g/L，硫酸铵3g/L，磷酸二氢钾0.5g/L，磷酸氢二钾1g/L，黄豆粉10g/L，玉米蛋白5g/L，玉米浆10g/L，鸟嘌呤0.1g/L，次黄嘌呤0.1g/L，维生素B1 01g/L，维生素B120.1g/L。
对照例：一种腺嘌呤生产菌种用培养基，它包含如下成分的组分：葡萄糖50g/L，硫酸铵5g/L，磷酸二氢钾1g/L，磷酸氢二钾1.5g/L，大豆蛋白胨25g/L，玉米浆10g/L。
菌种培养试验：
将12％腺嘌呤生产菌悬液接种到实施例1、2、3、4和对照四种配方中，在温度36±1℃条件下培养，检测菌液OD值、pH值、种龄、腺嘌呤含量，并在显微镜下观察菌体形态，培养试验结果如表1所示。
表1腺嘌呤菌种生长代谢数据
由表1数据可知，实例1、实例2、实例3、实例4的OD值、pH值达到或者优于对照例，达到移种的要求，且种龄比对照例缩短5-7小时，显微镜下观
本发明和现有技术相比的优点如下：
1)通过添加菌体生长所需的速效氮源酵母粉，缩短菌体生长延滞期，有利于菌体快速生长至对数生长期，从而缩短菌龄，增加了菌体活力。
2)对培养基中的其他氮源进行优选，寻找来源方便、价格低廉的原料进行替代，增加了配方的经济性和稳定性。
3)在配方中添加多种微量元素，丰富培养基组成，对菌体快速生产代谢所需的前体、辅酶进行补充，促进菌体合成和营养物质的代谢。
4)在本发明的配方实施下，腺嘌呤生产菌种所需的生长周期缩短，菌体活力强，且稳定性好。
实施方式
为了更好地展示上述技术方案，下面通过具体实例来对本发明的实施方式做进一步详细描述。
实施例1：一种腺嘌呤生产菌种用培养基，它包含如下成分的组分：葡萄糖50g/L，啤酒酵母浸粉5g/L，硫酸铵5g/L，磷酸二氢钾1g/L，磷酸氢二钾1.5g/L，黄豆粉10g/L，大米蛋白5g/L，玉米浆5g/L，生物素0.1g/L，次黄嘌呤0.1g/L，维生素B12 0.15g/L，维生素B10.1g/L。
实施例2：一种腺嘌呤生产菌种用培养基，它包含如下成分的组分：葡萄糖25g/L，面包酵母浸粉15g/L，硫酸铵5g/L，磷酸二氢钾1g/L，磷酸氢二钾1.5g/L，玉米蛋白5g/L，豆粕粉10g/L，玉米浆5g/L，生物素0.1g/L，鸟嘌呤0.15g/L，维生素B1 0.15g/L，维生素B120.1g/L。
实施例3：一种腺嘌呤生产菌种用培养基，它包含如下成分的组分：葡萄糖75g/L，面包酵母浸粉10g/L，硫酸铵3g/L，磷酸二氢钾0.5g/L，磷酸氢二察菌体形态均匀、整齐、数量多。故本发明培养基在腺嘌呤菌种生产中达到或优于预期标准。
菌种发酵试验：
将上述菌种培养试验得到的对照例菌液和实例1、实例2、实例3、实例4菌液分别接种于50L发酵罐培养，控制温度39±1℃，罐压0.05MPa，转速400-700rpm，培养周期50-54小时，检测最终发酵产物腺嘌呤含量，结果如表2所示。
表2 50L发酵罐腺嘌呤发酵试验数据
| 项目 | 对照例 | 实例1 | 实例2 | 实例3 | 实例4 |
| 发酵周期(小时 | 53 | 52 | 50 | 50 | 54 |
| 腺嘌呤含量(克/升) | 25.48 | 25.88 | 26.51 | 26.02 | 26.89 |
由表2数据可知，使用实例1、实例2、实例3、实例4培养基获得的菌液，接种到50L发酵罐后发酵生产的腺嘌呤含量达到或者优于对照培养基获得的菌液。
Claims (4)
1.一种腺嘌呤生产菌种用培养基，它包含如下含量的组分：葡萄糖10-100g/L，酵母粉5-15g/L，硫酸铵2-10g/L，磷酸二氢钾0.5-2.5g/L，磷酸氢二钾1-2.5g/L，其他氮源5-25g/L，玉米浆2-10g/L，微量元素0.2-1g/L。
2.根据权利要求1所述的腺嘌呤生产菌种用培养基，其特征在于：所述的酵母粉优选啤酒酵母浸粉和面包酵母浸粉。
3.根据权利要求1或2所述的腺嘌呤生产菌种用培养基，其特征在于：所述的其他氮源包括大豆蛋白胨、豆粕粉、黄豆粉、玉米蛋白、大米蛋白中的一种或者多种。
4.根据权利要求3所述的腺嘌呤生产菌种用培养基，其特征在于：所述的微量元素是指生物素、鸟嘌呤、次黄嘌呤、维生素B12、维生素B1中的多种。
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