Improving the refolding efficiency for proinsulin aspart inclusion body with optimized buffer compositions
文献类型：期刊论文
| 作者 | Chen, Ying1,2; Wang, Qi1,2; Zhang, Chun1; Li, Xiunan1; Gao, Qiang3; Dong, Changqing1; Liu, Yongdong1; Su, Zhiguo1 |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2016-06-01 |
| 卷号 | 122期号:JUNE页码:1-7 |
| 关键词 | Proinsulin Inclusion bodies Additive Oxidative refolding Diselenide Aggregate |
| ISSN号 | 1046-5928 |
| 英文摘要 | Successfully recovering proinsulin's native conformation from inclusion body is the crucial step to guarantee high efficiency for insulin's manufacture. Here, two by-products of disulfide-linked oligomers and disulfide-isomerized monomers were clearly identified during proinsulin aspart's refolding through multiple analytic methods. Arginine and urea are both used to assist in proinsulin refolding, however the efficacy and possible mechanism was found to be different. The oligomers formed with urea were of larger size than with arginine. With the urea concentrations increasing from 2 M to 4 M, the content of oligomers decreased greatly, but simultaneously the refolding yield at the protein concentration of 0.5 mg/mL decreased from 40% to 30% due to the increase of disulfide-isomerized monomers. In contrast, with arginine concentrations increasing up to 1 M, the refolding yield gradually increased to 50% although the content for oligomers also decreased. Moreover, it was demonstrated that not redox pairs but only oxidant was necessary to facilitate the native disulfide bonds formation for the reduced denatured proinsulin. An oxidative agent of selenocystamine could increase the yield up to 80% in the presence of 0.5 M arginine. Further study demonstrated that refolding with 2 M urea instead of 0.5 M arginine could achieve similar yield as protein concentration is slightly reduced to 0.3 mg/mL. In this case, refolded proinsulin was directly purified through one-step of anionic exchange chromatography, with a recovery of 32% and purity up to 95%. All the results could be easily adopted in insulin's industrial manufacture for improving the production efficiency. (C) 2016 Elsevier Inc. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | HUMAN INSULIN ; PROTEIN ; EXPRESSION ; DISELENIDES ; CATALYSTS ; CLEAVAGE ; ARGININE ; TRYPSIN ; PATHWAY |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000375172200001 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/21029]  |
| 专题 | 过程工程研究所_生化工程国家重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Novo Nordisk Res Ctr China, Beijing 102206, Peoples R China |
| 推荐引用方式 GB/T 7714 | Chen, Ying,Wang, Qi,Zhang, Chun,et al. Improving the refolding efficiency for proinsulin aspart inclusion body with optimized buffer compositions[J]. PROTEIN EXPRESSION AND PURIFICATION,2016,122(JUNE):1-7. |
| APA | Chen, Ying.,Wang, Qi.,Zhang, Chun.,Li, Xiunan.,Gao, Qiang.,...&Su, Zhiguo.(2016).Improving the refolding efficiency for proinsulin aspart inclusion body with optimized buffer compositions.PROTEIN EXPRESSION AND PURIFICATION,122(JUNE),1-7. |
| MLA | Chen, Ying,et al."Improving the refolding efficiency for proinsulin aspart inclusion body with optimized buffer compositions".PROTEIN EXPRESSION AND PURIFICATION 122.JUNE(2016):1-7. |
入库方式： OAI收割
来源：过程工程研究所
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