Artificial chaperone-assisted refolding in a microchannel.
Protein refolding using a simple dilution method in a microchannel often led to the formation of protein aggregates, which bound to the microchannel wall, resulting in low refolding yields. To inhibit aggregation and improve refolding yields, an artificial chaperone-assisted (ACA) refolding, which employed detergents and beta-cyclodextrin was used. Model proteins, hen egg white lysozyme and yeast alpha-glucosidase, were successfully refolded in a microchannel. The microscopic observation showed that the ACA method suppressed protein aggregation and facilitated the refolding of lysozyme, whereas significant aggregation was observed when a simple dilution method was employed. The ACA method increased the lysozyme refolding yield by 40% over the simple dilution approach. Similarly, for a-glucosidase, the refolding yield using the ACA method (ca. 50%) was approximately three times compared with the simple dilution method. The ACA refolding method is a suitable approach to use in the refolding of proteins using a microfluidic system.
Yamamoto
Etsushi
E
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Yamaguchi
Satoshi
S
Sasaki
Naoki
N
Kim
Haeng-Boo
HB
Kitamori
Takehiko
T
Nagamune
Teruyuki
T
D000818:Animals; D002645:Chickens; D003902:Detergents / Q000737:chemistry; D004527:Egg Proteins / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D009113:Muramidase / Q000737:chemistry*; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D000520:alpha-Glucosidases / Q000737:chemistry*; D047392:beta-Cyclodextrins / Q000737:chemistry
D003902:Detergents; D004527:Egg Proteins; D029701:Saccharomyces cerevisiae Proteins; D047392:beta-Cyclodextrins; D009113:Muramidase; D000520:alpha-Glucosidases