胶质芽孢杆菌PM13菌株发酵工艺的研究
文献类型：学位论文
| 作者 | 王雪 |
| 学位类别 | 硕士 |
| 答辩日期 | 2010-05-24 |
| 授予单位 | 中国科学院研究生院 |
| 导师 | 袁晓凡 |
| 关键词 | 胶质芽孢杆菌PM13菌株 多糖组成 培养基成分 培养条件 发酵工艺 |
| 其他题名 | Study on the fermentation technology of bacillus mucilaginosus PM13 strain |
| 学位专业 | 生物化工 |
| 中文摘要 | 本文对一种新诱变胶质芽孢杆菌菌株PM13的培养基成分和培养条件进行了优化，确定了该菌株的培养工艺，使其芽孢生成量得到显著提高的同时显著降低了发酵液粘度；对该菌株所产胞外多糖组成进行了研究，确定了胞外多糖的单糖组成，为PM13菌株的工业化生产和土壤改良机理研究奠定基础。本论文的主要研究结果如下： 对PM13菌株生长特性进行了研究，在进行一代活化后该菌可以获得良好的传代性能，生长周期为40 h，可以用于工业连续化生产。 通过气相色谱分析得知PM13菌株所产胞外多糖中含有葡萄糖、半乳糖、甘露糖和木糖，其中前三者为主要成分，各种糖的含量比例为葡萄糖:甘露糖:半乳糖:木糖=274:377:209:70。 通过单因素实验、正交实验和均匀实验得到PM13菌株的最佳培养基成分（g/L）为：糖蜜 3.67、淀粉 1.83、豆粕粉 7.0、CaCO3 8.5、K2HPO4 2.0、MgSO4•7H2O 1.4、NaCl 0.2，芽孢生成量为2.2×108 cfu/mL，与未优化时比较，芽孢生成量提高了105倍，并且发酵液稠度系数显著降低到0.028 Pasn。 通过单因素实验得到PM13菌株优化后培养条件为：初始pH值7.5、温度34℃、转速200 r/min、接种量3%、装液量50 mL/250 mL三角瓶，在此条件下培养的芽孢生成量为1.2×109 cfu/mL，比未优化前提高了5倍，并且发酵液稠度系数为0.015 Pasn。 通过PM13菌株在10 L机械搅拌式生物反应器中进行发酵的研究得知，在消泡剂添加量0.03%、搅拌转速前期250 r/min、后期500 r/min，装液量70%，接种量5%的发酵工艺下，经过36 h的发酵，可以获得1.0×109 cfu/mL芽孢生成量，并且在整个发酵过程中发酵液稠度系数不高于0.21 Pasn。 |
| 英文摘要 | The culture technology of a newly mutagenetic Bacillus mucilaginosus strain PM13 was investigated by optimizing the culture medium and culture conditions in order to increase the production of spores and decrease the viscosity of the broth. The monosaccharide composition of polysaccharide produced by PM13 strain was also detected in order to lay a foundation for the study on the mechanism of its soil improvement. The main experimental results were as follows: The characteristics of PM13 strain growth were investigated. Experimental results showed that PM13 strain had good passage performance after activated one generation and the growth cycle was 40 h. So the PM13 strain can be used in the continuous production. The results of the gas chromatography analysis showed that the polysaccharide produced by PM13 strain was mainly composed of glucose, galactose, mannose and xylose with the ratio of 274:377:209:70. The optimized culture medium (g/L) of PM13 strain determined through single factor, orthogonal and uniform experiments was as follows: molasses 3.67, starch 1.83, soybean powder 7.0, CaCO3 8.5, K2HPO4 2.0, MgSO4•7H2O 1.4, NaCl 0.2. With this culture medium the production of spores reached 2.2×108 cfu/mL, which was 105 times of that unoptimized, and the fluid consistency coefficient of the broth decreased to 0.028 Pasn. The optimized culture conditions of PM13 strain optimized through single factor experiments were as follows: initial pH 7.5, temperature 34℃, rotation speed 200 r/min, inoculation 3%, and medium load was 50 mL in a 250 mL flask. Under these conditions, the production of spores reached 1.2×109 cfu/mL, which was 5 times of that under unoptimized conditions, and the fluid consistency coefficient of the broth was 0.015 Pasn. The fermentation of PM13 strain in a 10 L mechanically stirring bioreactor was studied. The fermentation technology was as follows: the concentratrion of antifoam agent was 0.03%, rotation speed was 250 r/min in initial stage and 500 r/min in later stage, medium load was 70% and inoculation was 5%. Under these conditions, the production of spores was 1.0×109 cfu/mL after 36 h fermentation, and the fluid consistency coefficient of the broth was no more than 0.21 Pasn during the whole fermentation. |
| 语种 | 中文 |
| 公开日期 | 2013-09-22 |
| 页码 | 96 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/1617]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 推荐引用方式 GB/T 7714 | 王雪. 胶质芽孢杆菌PM13菌株发酵工艺的研究[D]. 中国科学院研究生院. 2010. |
入库方式： OAI收割
来源：过程工程研究所
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