重组毕赤酵母胸腺肽α1-人血清白蛋白基因工程菌高密度发酵及分离纯化  

High cell density fermentation and purification of recombinant Pichia Pastoris Tα1-HSA genetically engineered bacteria

作　　者：马娇颖[1] 章成昌[1] 仇黎鹏[1] 姜玉涛[1] 闫璐颖[1] 陈菁[1] 陈建华[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《中国生化药物杂志》2012年第6期720-724,共5页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的研究重组毕赤酵母胸腺肽α1-人血清白蛋白(Tα1-HSA)基因工程菌在30 L发酵罐中高密度发酵表达Tα1-HSA融合蛋白的发酵工艺及产物的分离纯化方法。方法采用两步发酵法进行重组毕赤酵母Tα1-HSA基因工程菌的高密度发酵。发酵液经超滤浓缩,阴离子交换色谱,亲和色谱,凝胶过滤色谱等步骤进行分离纯化。结果发酵结束后,菌体细胞湿重达到350 g/L,发酵液中蛋白产量达到2.0 g/L。发酵液经分离纯化后获得了纯度较高的重组Tα1-HSA融合蛋白,HPLC分析纯度达97.5%。结论重组毕赤酵母Tα1-HSA基因工程菌高密度发酵的成功及分离纯化方法的建立为Tα1-HSA的进一步研究和开发奠定了基础。Purpose To study the high cell density fermentation of recombinant Pichia Pastoris Tα- HSA genetically engineered bacteria, and the purification of expressed target protein. Methods A two-step fermentation is taken for the high cell density fermentation of recombinant Pichia Pastoris Tal-HSA genet- ically engineered bacteria. Tα-HSA was purified from fermentation broth by uhrafihration,anion excha-nge chromatography,affinity chromatography and gel filtration chromatography. Results Through the con- trol and optimization of fermentation conditions, the bacterial cell wet weight could reach 350 g/L and the productivity of expressed Tα-HSA could reach 2.0 g/l. in 30 l, fermentor. High purity recombinant Tal- of HSA fusion protein is obtained after separation and purification. The HPLC analysis shows that the pu- rified Tα-HSA has an overall purity of 97.5%. Conclusion The success of the high cell density fermen- tation and purification has laid a foundation for the further research and development of Tα1-HSA.

关 键 词：胸腺肽Α1 人血清白蛋白 毕赤酵母 高密度发酵 分离纯化 

分 类 号：TQ929[轻工技术与工程—发酵工程] Q78[生物学—分子生物学]