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Laccase mediator system for activation of agarose gel: Application for immobilization of proteins

Mendoza, Laura LU ; Mamo, Gashaw LU ; Flores, Ninoska; Gimenez, Alberto and Hatti-Kaul, Rajni LU (2011) In Journal of Molecular Catalysis B: Enzymatic 68(3-4). p.270-274
Abstract
Cross-linked Sepharose beads were treated with laccase-TEMPO system for oxidation of the primary alcohol groups on the sugar moieties. Optimal activation conditions using Trametes versicolor laccase were at pH 5 and 22 degrees C, giving an aldehyde content of 55 mu mol g(-1) Sepharose with 28 units g(-1) of laccase and 12.5 mM TEMPO. The activated Sepharose was used for immobilization of trypsin as model protein. Highest degree of immobilization was obtained at pH 10.5 but the activity yield was only 31% of that loaded on the gel. The yield of gel bound trypsin activity was increased to 76% (corresponding to about 43 U g(-1) Sepharose) when the immobilization was performed in the presence of trypsin inhibitor, benzamidine. The... (More)
Cross-linked Sepharose beads were treated with laccase-TEMPO system for oxidation of the primary alcohol groups on the sugar moieties. Optimal activation conditions using Trametes versicolor laccase were at pH 5 and 22 degrees C, giving an aldehyde content of 55 mu mol g(-1) Sepharose with 28 units g(-1) of laccase and 12.5 mM TEMPO. The activated Sepharose was used for immobilization of trypsin as model protein. Highest degree of immobilization was obtained at pH 10.5 but the activity yield was only 31% of that loaded on the gel. The yield of gel bound trypsin activity was increased to 76% (corresponding to about 43 U g(-1) Sepharose) when the immobilization was performed in the presence of trypsin inhibitor, benzamidine. The immobilization yields were comparable to that obtained on the matrix activated using sodium periodate (containing 72 mu mol aldehyde per g Sepharose). Recycling and storage of the immobilized trypsin preparations showed high stability of the enzyme bound to laccase-TEMPO activated gel. (C) 2010 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Laccase mediator system, Trametes versicolor, Cross-linked Sepharose, Periodate activation, Trypsin
in
Journal of Molecular Catalysis B: Enzymatic
volume
68
issue
3-4
pages
270 - 274
publisher
Elsevier
external identifiers
  • wos:000287114900008
  • scopus:78651385759
ISSN
1873-3158
DOI
10.1016/j.molcatb.2010.11.016
language
English
LU publication?
yes
id
a2951951-b008-4d55-80e2-9328818960dc (old id 1877978)
date added to LUP
2011-04-15 15:24:38
date last changed
2017-07-02 03:23:12
@article{a2951951-b008-4d55-80e2-9328818960dc,
  abstract     = {Cross-linked Sepharose beads were treated with laccase-TEMPO system for oxidation of the primary alcohol groups on the sugar moieties. Optimal activation conditions using Trametes versicolor laccase were at pH 5 and 22 degrees C, giving an aldehyde content of 55 mu mol g(-1) Sepharose with 28 units g(-1) of laccase and 12.5 mM TEMPO. The activated Sepharose was used for immobilization of trypsin as model protein. Highest degree of immobilization was obtained at pH 10.5 but the activity yield was only 31% of that loaded on the gel. The yield of gel bound trypsin activity was increased to 76% (corresponding to about 43 U g(-1) Sepharose) when the immobilization was performed in the presence of trypsin inhibitor, benzamidine. The immobilization yields were comparable to that obtained on the matrix activated using sodium periodate (containing 72 mu mol aldehyde per g Sepharose). Recycling and storage of the immobilized trypsin preparations showed high stability of the enzyme bound to laccase-TEMPO activated gel. (C) 2010 Elsevier B.V. All rights reserved.},
  author       = {Mendoza, Laura and Mamo, Gashaw and Flores, Ninoska and Gimenez, Alberto and Hatti-Kaul, Rajni},
  issn         = {1873-3158},
  keyword      = {Laccase mediator system,Trametes versicolor,Cross-linked Sepharose,Periodate activation,Trypsin},
  language     = {eng},
  number       = {3-4},
  pages        = {270--274},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Catalysis B: Enzymatic},
  title        = {Laccase mediator system for activation of agarose gel: Application for immobilization of proteins},
  url          = {http://dx.doi.org/10.1016/j.molcatb.2010.11.016},
  volume       = {68},
  year         = {2011},
}