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Immobilization of thermostable beta-glucosidase variants on acrylic supports for biocatalytic processes in hot water

Khan, Sami LU ; Lindahl, Sofia LU ; Turner, Charlotta LU and Nordberg Karlsson, Eva LU (2012) In Journal of Molecular Catalysis B: Enzymatic 80. p.28-38
Abstract
Two variants of the thermostable beta-glucosidase TnBgl1A (wt and N221S/P342L) from Thermotoga neapolitana were immobilized on acrylic supports (Eupergit (R) C, Eupergit (R) C250L, and cryogel) and evaluated at conditions close to the boiling point of water. Thermo-gravimetric analysis showed the supports to be stable <250 degrees C. Both wt and N221S/P342L were covalently bound to oxirane-groups respectively via glutaraldehyde spacers, and for coupling reactions 26 Lys and 20 Ser/Thr were surface-located. Immobilized enzymes were active on all supports in the temperature range 80-95 degrees C, but the observed specific activity was low (<= 19 U mg(-1)) using the cryogel. More than 91% of the initial activity was maintained after ten... (More)
Two variants of the thermostable beta-glucosidase TnBgl1A (wt and N221S/P342L) from Thermotoga neapolitana were immobilized on acrylic supports (Eupergit (R) C, Eupergit (R) C250L, and cryogel) and evaluated at conditions close to the boiling point of water. Thermo-gravimetric analysis showed the supports to be stable <250 degrees C. Both wt and N221S/P342L were covalently bound to oxirane-groups respectively via glutaraldehyde spacers, and for coupling reactions 26 Lys and 20 Ser/Thr were surface-located. Immobilized enzymes were active on all supports in the temperature range 80-95 degrees C, but the observed specific activity was low (<= 19 U mg(-1)) using the cryogel. More than 91% of the initial activity was maintained after ten times recycling, and the same was recovered after 3 months storage at 4 degrees C for Eupergit (R) supports by simply incubating the preparation with bovine serum albumin. No storage loss was detectable on cryogels. The glutaraldehyde spacer improved activity on cryogels, but not on Eupergie supports. Immobilization on Eupergit (R) C250L yielded the highest observed specific activity (254 U mg(-1) for N221S/P342L) in a procedure including blocking of free oxirane-groups by BSA. This biocatalyst was used for on-line hydrolysis of quercetin-glucosides in a yellow onion extract at 80 degrees C, proving the immobilized biocatalyst to be promising in on-line systems for extraction and hydrolysis using hot pressurized water. (C) 2012 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Immobilization, Glycoside hydrolase family 1, Eupergit (R) C, Eupergit, (R) C250L, Cryogel
in
Journal of Molecular Catalysis B: Enzymatic
volume
80
pages
28 - 38
publisher
Elsevier
external identifiers
  • wos:000305866100005
  • scopus:84860831353
ISSN
1873-3158
DOI
10.1016/j.molcatb.2012.01.004
language
English
LU publication?
yes
id
58a9c0eb-33d5-48f5-9cfd-e9da0dbca7ec (old id 3001359)
date added to LUP
2012-08-21 12:37:03
date last changed
2017-10-01 03:01:50
@article{58a9c0eb-33d5-48f5-9cfd-e9da0dbca7ec,
  abstract     = {Two variants of the thermostable beta-glucosidase TnBgl1A (wt and N221S/P342L) from Thermotoga neapolitana were immobilized on acrylic supports (Eupergit (R) C, Eupergit (R) C250L, and cryogel) and evaluated at conditions close to the boiling point of water. Thermo-gravimetric analysis showed the supports to be stable &lt;250 degrees C. Both wt and N221S/P342L were covalently bound to oxirane-groups respectively via glutaraldehyde spacers, and for coupling reactions 26 Lys and 20 Ser/Thr were surface-located. Immobilized enzymes were active on all supports in the temperature range 80-95 degrees C, but the observed specific activity was low (&lt;= 19 U mg(-1)) using the cryogel. More than 91% of the initial activity was maintained after ten times recycling, and the same was recovered after 3 months storage at 4 degrees C for Eupergit (R) supports by simply incubating the preparation with bovine serum albumin. No storage loss was detectable on cryogels. The glutaraldehyde spacer improved activity on cryogels, but not on Eupergie supports. Immobilization on Eupergit (R) C250L yielded the highest observed specific activity (254 U mg(-1) for N221S/P342L) in a procedure including blocking of free oxirane-groups by BSA. This biocatalyst was used for on-line hydrolysis of quercetin-glucosides in a yellow onion extract at 80 degrees C, proving the immobilized biocatalyst to be promising in on-line systems for extraction and hydrolysis using hot pressurized water. (C) 2012 Elsevier B.V. All rights reserved.},
  author       = {Khan, Sami and Lindahl, Sofia and Turner, Charlotta and Nordberg Karlsson, Eva},
  issn         = {1873-3158},
  keyword      = {Immobilization,Glycoside hydrolase family 1,Eupergit (R) C,Eupergit,(R) C250L,Cryogel},
  language     = {eng},
  pages        = {28--38},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Catalysis B: Enzymatic},
  title        = {Immobilization of thermostable beta-glucosidase variants on acrylic supports for biocatalytic processes in hot water},
  url          = {http://dx.doi.org/10.1016/j.molcatb.2012.01.004},
  volume       = {80},
  year         = {2012},
}