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Optimizing refolding and recovery of active recombinant Bacillus halodurans xylanase in polymer-salt aqueous two-phase system using surface response analysis

Rahimpour, Farshad LU ; Mamo, Gashaw LU ; Feyzi, Farzaneh; Maghsoudi, Saeid and Hatti-Kaul, Rajni LU (2007) In Journal of Chromatography A 1141(1). p.32-40
Abstract
An experimental design was used to determine optimal conditions for refolding of a recombinant thermostable and alkaline active xylanase from Bacillus halodurans in PEG-phosphate two-phase system. The influence of different experimental variables on the enzyme recovery has been evaluated. To build the mathematical model and minimize the number of experiments for the design parameters, response surface methodology with a face-centered central composite design (CCF) was defined based on the conditions found by preliminary tests that resulted in the highest refolding yield. The adequacy of the calculated model for the response was confirmed by means of variance analysis and additional experiments. Analysis of contours of constant response as... (More)
An experimental design was used to determine optimal conditions for refolding of a recombinant thermostable and alkaline active xylanase from Bacillus halodurans in PEG-phosphate two-phase system. The influence of different experimental variables on the enzyme recovery has been evaluated. To build the mathematical model and minimize the number of experiments for the design parameters, response surface methodology with a face-centered central composite design (CCF) was defined based on the conditions found by preliminary tests that resulted in the highest refolding yield. The adequacy of the calculated model for the response was confirmed by means of variance analysis and additional experiments. Analysis of contours of constant response as a function of pH, polyethylene glycol (PEG) molecular weight and concentration, and salt concentration for different enzyme loads revealed different effects of these five factors on the studied parameters. Recovery of more than 92% active xylanase was predicted for a system with 18.3% (w/w) PEG 1000, 14.4% (w/w) phosphate at pH 8.5, and enzyme load corresponding to a protein concentration of about 0.05 mg/g system. The yield of the refolded enzyme was found to be optimal at 22 degrees C. The validity of the response model was verified by a good agreement between predicted and experimental results. (c) 2006 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
composite design, central, PEG-phosphate aqueous two-phase systems, refolding, xylanase
in
Journal of Chromatography A
volume
1141
issue
1
pages
32 - 40
publisher
Elsevier
external identifiers
  • wos:000243864100004
  • scopus:33846008444
ISSN
0021-9673
DOI
10.1016/j.chroma.2006.11.053
language
English
LU publication?
yes
id
6e404607-e985-46b6-a441-f45e2874c051 (old id 676182)
date added to LUP
2007-12-20 08:31:19
date last changed
2017-09-10 04:34:13
@article{6e404607-e985-46b6-a441-f45e2874c051,
  abstract     = {An experimental design was used to determine optimal conditions for refolding of a recombinant thermostable and alkaline active xylanase from Bacillus halodurans in PEG-phosphate two-phase system. The influence of different experimental variables on the enzyme recovery has been evaluated. To build the mathematical model and minimize the number of experiments for the design parameters, response surface methodology with a face-centered central composite design (CCF) was defined based on the conditions found by preliminary tests that resulted in the highest refolding yield. The adequacy of the calculated model for the response was confirmed by means of variance analysis and additional experiments. Analysis of contours of constant response as a function of pH, polyethylene glycol (PEG) molecular weight and concentration, and salt concentration for different enzyme loads revealed different effects of these five factors on the studied parameters. Recovery of more than 92% active xylanase was predicted for a system with 18.3% (w/w) PEG 1000, 14.4% (w/w) phosphate at pH 8.5, and enzyme load corresponding to a protein concentration of about 0.05 mg/g system. The yield of the refolded enzyme was found to be optimal at 22 degrees C. The validity of the response model was verified by a good agreement between predicted and experimental results. (c) 2006 Elsevier B.V. All rights reserved.},
  author       = {Rahimpour, Farshad and Mamo, Gashaw and Feyzi, Farzaneh and Maghsoudi, Saeid and Hatti-Kaul, Rajni},
  issn         = {0021-9673},
  keyword      = {composite design,central,PEG-phosphate aqueous two-phase systems,refolding,xylanase},
  language     = {eng},
  number       = {1},
  pages        = {32--40},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Optimizing refolding and recovery of active recombinant Bacillus halodurans xylanase in polymer-salt aqueous two-phase system using surface response analysis},
  url          = {http://dx.doi.org/10.1016/j.chroma.2006.11.053},
  volume       = {1141},
  year         = {2007},
}