Rapid renaturation of denatured and aggregated proteins using liquid paraffin as a pseudolipid bilayer membrane.

Abstract:

:A means of rapidly renaturing denatured protein was devised and evaluated. Three liquids were laminarly layered in a centrifuge tube, in which two solutions sandwiched liquid paraffin so as to form a pseudolipid bilayer. Denatured and aggregted protein placed on the upper surface of liquid paraffin was renatured as it passed through liquid-paraffin layer into the renaturation buffer during the centrifugation. The aggregated and denatured protein selectively passed through the liquid-paraffin layer, whereas other solutions, such as chaotropic agents or organic solvent, could not. This means that a rapid dilution condition favorable for protein renaturation was realized in a small scale. Aggregated and denatured BSA and ribonuclease A were renatured and resolubilized as they passed through the liquid-paraffin layer into an appropriate renaturation buffer solution. This method was also applied to the rapid heme reconstitution of myoglobin from Feprotoporphyrin IX to Zn-protoporphyrin IX. (c) 1994 John Wiley & Sons, Inc.

journal_name

Biotechnol Bioeng

authors

Yoshii H,Kojima T,Furuta T,Yasunishi A

doi

10.1002/bit.260430108

subject

Has Abstract

pub_date

1994-01-05 00:00:00

pages

57-63

issue

1

eissn

0006-3592

issn

1097-0290

journal_volume

43

pub_type

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