Abstract
This article reports that a reversed micellar solution is useful for refolding proteins directly from a solid source. The solubilization of denatured RNase A, which had been prepared by reprecipitation from the denaturant protein solution, into reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) has been investigated by s solid-liquid extraction system. This method is an alternative to the ordinary protein extraction in reversed micelles based on the liquid-liquid extraction. The solid-liquid extraction method was found to facilitate the solubilization of denatured proteins more efficiently in the reversed micellar media than the ordinary phase transfer method of liquid extraction. The refolding of denatured RNase A entrapped in reversed micelles was attained by adding a redox reagent (reduced and oxidized glutathion). Enzymatic activity of RNase A was gradually recovered with time in the reversed micelles. The denatured RNase A was completely refolded within 30 h. In addition, the efficiency of protein refolding was enhanced when reversed micelles were applied to denatured RNase A containing a higher protein concentration that, in the case of aqueous media, would lead to protein aggregation. The solid-liquid extraction technique using reversed micelles affords better scale-up advantages in the direct refolding process of insoluble protein aggregates.
| Original language | English |
|---|---|
| Pages (from-to) | 620-623 |
| Number of pages | 4 |
| Journal | Biotechnology and Bioengineering |
| Volume | 57 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - Mar 5 1998 |
Fingerprint
All Science Journal Classification (ASJC) codes
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
Cite this
Protein refolding by reversed micelles utilizing solid-liquid extraction technique. / Hashimoto, Yukihisa; Ono, Tsutomu; Goto, Masahiro; Hatton, T. Alan.
In: Biotechnology and Bioengineering, Vol. 57, No. 5, 05.03.1998, p. 620-623.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Protein refolding by reversed micelles utilizing solid-liquid extraction technique
AU - Hashimoto, Yukihisa
AU - Ono, Tsutomu
AU - Goto, Masahiro
AU - Hatton, T. Alan
PY - 1998/3/5
Y1 - 1998/3/5
N2 - This article reports that a reversed micellar solution is useful for refolding proteins directly from a solid source. The solubilization of denatured RNase A, which had been prepared by reprecipitation from the denaturant protein solution, into reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) has been investigated by s solid-liquid extraction system. This method is an alternative to the ordinary protein extraction in reversed micelles based on the liquid-liquid extraction. The solid-liquid extraction method was found to facilitate the solubilization of denatured proteins more efficiently in the reversed micellar media than the ordinary phase transfer method of liquid extraction. The refolding of denatured RNase A entrapped in reversed micelles was attained by adding a redox reagent (reduced and oxidized glutathion). Enzymatic activity of RNase A was gradually recovered with time in the reversed micelles. The denatured RNase A was completely refolded within 30 h. In addition, the efficiency of protein refolding was enhanced when reversed micelles were applied to denatured RNase A containing a higher protein concentration that, in the case of aqueous media, would lead to protein aggregation. The solid-liquid extraction technique using reversed micelles affords better scale-up advantages in the direct refolding process of insoluble protein aggregates.
AB - This article reports that a reversed micellar solution is useful for refolding proteins directly from a solid source. The solubilization of denatured RNase A, which had been prepared by reprecipitation from the denaturant protein solution, into reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) has been investigated by s solid-liquid extraction system. This method is an alternative to the ordinary protein extraction in reversed micelles based on the liquid-liquid extraction. The solid-liquid extraction method was found to facilitate the solubilization of denatured proteins more efficiently in the reversed micellar media than the ordinary phase transfer method of liquid extraction. The refolding of denatured RNase A entrapped in reversed micelles was attained by adding a redox reagent (reduced and oxidized glutathion). Enzymatic activity of RNase A was gradually recovered with time in the reversed micelles. The denatured RNase A was completely refolded within 30 h. In addition, the efficiency of protein refolding was enhanced when reversed micelles were applied to denatured RNase A containing a higher protein concentration that, in the case of aqueous media, would lead to protein aggregation. The solid-liquid extraction technique using reversed micelles affords better scale-up advantages in the direct refolding process of insoluble protein aggregates.
UR - http://www.scopus.com/inward/record.url?scp=0032485469&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032485469&partnerID=8YFLogxK
U2 - 10.1002/(SICI)1097-0290(19980305)57:5<620::AID-BIT14>3.0.CO;2-F
DO - 10.1002/(SICI)1097-0290(19980305)57:5<620::AID-BIT14>3.0.CO;2-F
M3 - Article
C2 - 10099241
AN - SCOPUS:0032485469
VL - 57
SP - 620
EP - 623
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
SN - 0006-3592
IS - 5
ER -