Refolding of Denatured and Reduced Lysozyme with Cysteine/Cystine Red/Ox Solution in Diafiltration

JOURNAL OF CHEMICAL ENGINEERING OF JAPAN

TAKAHIRO YONEHARA¹  PEKKA LINKO²  TAKESHI FURUTA¹  DAISUKE ITO¹  HIDEFUMI YOSHII¹ 

[1] Department of Biotechnology, Tottori University;Department of Chemical Technology, Helsinki University of Technology

关键词: Refolding;    Lysozyme;    Diafiltration;    Cysteine;    Cystine;   

DOI  :  10.1252/jcej.34.211

来源: Maruzen Company Ltd

【 摘 要 】

References(7)Cited-By(3)Refolding of reduced and denatured protein in vitro has been an important issue for both basic research and applied biotechnology. Refolding at low protein concentration requires large volumes of refolding buffer. Diafiltration method is useful to control the denaturant and red/ox reagents in the refolding solution. We constructed a refolding procedure for high concentrations of reduced and denatured lysozyme of about 10 mg/ml (700 μM) on linear reduction of urea concentration in diafiltration, 0.8 mM cystine and 8 mM cysteine under nitrogen at 2 atm. This method can obtain about 90% refolding yield at 700 μM and almost 100% in 350 μM lysozyme. The refolding yields during the diafiltration can be simulated using the competitive reaction between the refolding and aggregation. In the red/ox control with cysteine and cystine, a rate order of aggregation reaction of near 2 is obtained.

【 授权许可】

Unknown