Enhanced metal-affinity partitioning of genetically engineered hirudin variants in polyethylene glycol/dextran two-phase systems

Cited 4 time in scopus
Metadata Downloads
Title
Enhanced metal-affinity partitioning of genetically engineered hirudin variants in polyethylene glycol/dextran two-phase systems
Author(s)
Bong Hyun Chung; Jung Hoon Sohn; Sang Ki Rhee; Y K Chang; Young Hoon Park
Bibliographic Citation
Journal of Fermentation Bioengineering, vol. 77, no. 1, pp. 75-79
Publication Year
1994
Abstract
Hirudin variants were constructed to exhibit an increased metal-binding affinity in an attempt to apply a metal-affinity partitioning process in a primary separation step for purification of hirudin. The hirudin variants were genetically engineered to contain additional surface-accessible histidines and produced by recombinant Saccharomyces cerevisiae. The partitioning behavior of these variants was compared with that of the wild type with a single surface-accessible histidine at position 51. Upon the addition of a small amount of Cu(II)IDA-PEG (Cu(II)iminodiacetic acid-polyethylene glycol) ligand to PEG/dextran two-phase systems, the hirudin variants with two or three surface-accessible histidines were more selectively partitioned into the PEG-rich phase than the wild type. Integrating protein engineering to metal-affinity partitioning offers the potential for general application of this technique to facilitate protein isolation, but the genetically engineered protein variants should be carefully constructed in a manner to minimize reduction of native protein activity.
ISSN
0385-6380
Publisher
Soc Bioscience Bioengineering Japan
DOI
http://dx.doi.org/10.1016/0922-338X(94)90212-7
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.