Development of an effective folding procedure for SS-containing proteins by using a micro flow chemistry

2015 Fiscal Year Final Research Report

• Project/Area Number: 26888016
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Bio-related chemistry
• Research Institution: Tokai University
• Principal Investigator: Arai Kenta 東海大学, 理学部, 助教 (60728062)
• Project Period (FY): 2014-08-29 – 2016-03-31
• Keywords: フォールディング / マイクロリアクター / ジスルフィド / フローケミストリー / インスリン / リゾチーム

Outline of Final Research Achievements

A chemically or biologically synthesized polypeptide needs to fold into the bioactive native form through random disulfide (SS) formation and SS isomerization. In this research, development of a facile methodology, which allows any scientists in various fields to achieve an effective folding, was attempted only by using a micro flow reactor (MR), which enables a rapid reaction due to the very short diffusion distances, without special organic reagents. When the each solution of reduced lysozyme as a model protein and glutathione was flowed into the MR at the same time, the bioactive lysozyme was gradually recovered. It is worth noting that the refolding velocity under a flow condition was faster than that under a batch condition. Similar folding behavior was observed for the folding of double-chain insulin coupled with inter-chain SS formation. In conclusion, the micro flow technology has enabled the effective oxidative folding reaction without a complicating methodology.

Free Research Field

生物物理化学