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混合反胶团萃取分离蛋白质
严勇朝
Subtype博士
Thesis Advisor陈家镛
1997-10-01
Degree Grantor中国科学院研究生院
Abstract以分子量较大的牛血红蛋白、α - 淀粉酶和分子量较小的细胞色素C、α- 糜蛋白酶为模型蛋白,将非离子萃取剂和表面活性剂以及阳离子表面活性剂与阴离子表面活性剂AOT混合形成反胶团,系统考察了混合反胶团与单一反胶团萃取性能的差别、反胶团的微观结构以及蛋白质在反胶团中的构象变化。从混合反胶团体系中遴选出对目标酶有良好萃取性能且较好地保存酶活性的体系,直接从发酵液中分离提纯目标酶。实验结果表明,萃取剂、非离子表面活性剂与AOT混合能形成比AOT反胶团粒径更大的反胶团;非离子萃取剂和表面活性剂的加入不影响AOT反胶团的形貌;所研究的大多数混合反胶团体系对模型蛋白比单一反胶团体系有更好的萃取性能。对于特定的混合反胶团体系,蛋白质的反萃率和酶活回收率比单一AOT体系要高。不仅如此,蛋白质在特定的混合反胶团中比在单一AOT反胶团中变性程度要小,保持原有构象能力更强。在优化的萃取和反萃条件下,优选出的选择性和萃取分离效果均好且较好地保持酶活性的混合体系,可初步达到直接从发酵液中分离α- 淀粉酶的目的,且效果比用单一AOT体系好。
Other AbstractIn this paper, we tried to use the reversed micelles, (RM) formed by mixing nonionic extractant or nonionic cationic surfactant with AOT in isooctane to extrtact forur proteins with different molecular weight and pI. They are α-chymotrypsin and cytochrome C. A systematic study on the performance of protein extraction by the mixed Rms, the microstructure of Rms and the effect of Rms on the secondary structure of proteins studied were carried out. The mixed RMs with the best extraction performance and the least secondary conformation variations of proteins were chosen to purify the α-amylase directly from the industrial fermentation broth. Dynamic Light-Scattering study show that the size of the Rms formed by mixing nonionic extractant and surfactant with AOT is larger than that formed by AOT only. Freeze-Fracture Transmission Electron Microscopy study show that the addition of nonionic extractant and surfactant to AOT RM do not influence the shape of AOT RM. Better protein extraction results with the mixed Rms were obtained in comparison with AOT Rms. Under the same conditions, the percentage of back-extraction and remanined activity obtained by special mixed Rms is much higher than those obtained by AOT RM. The UV and Fourier Transform Infrared spectra studies show that lower changes of the conformation of proteins in nonionic extractant surfatant-AOT RM the those in AOT RM. Under the optimum extraction and back-extraction conditions, it is possible for SE10-AOT RM with the best extraction performance and lowest conformation changes of α-amylase to separate and purify α-amylase directly from the industrial fermentation broth with required pretreatment.
Pages153
Language中文
Document Type学位论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/8339
Collection研究所(批量导入)
Recommended Citation
GB/T 7714
严勇朝. 混合反胶团萃取分离蛋白质[D]. 中国科学院研究生院,1997.
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