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Fusion of carbohydrate binding modules from Thermotoga neapolitana with a family 10 xylanase from Bacillus halodurans S7

Mamo, Gashaw LU ; Hatti-Kaul, Rajni LU and Mattiasson, Bo LU (2007) In Extremophiles 11(1). p.169-177
Abstract
Xylanase A of Thermotoga neapolitana contains binding domains both at the N- and C-terminal ends of the catalytic domain. In the N-terminal position it contains two carbohydrate-binding modules (CBM) which belong to family 22. These CBMs bind xylan but not to cellulose. The gene encoding the mature peptide of these CBMs was fused with an alkaline active GH10 xylanase from Bacillus halodurans S7 and expressed in Escherichia coli. The (His)(6) tagged hybrid protein was purified by immobilized metal affinity chromatography and characterized. Xylan binding by the chimeric protein was influenced by NaCl concentration and pH of the binding medium. Binding increased with increasing salt concentration up to 200 mM. Higher extent of binding was... (More)
Xylanase A of Thermotoga neapolitana contains binding domains both at the N- and C-terminal ends of the catalytic domain. In the N-terminal position it contains two carbohydrate-binding modules (CBM) which belong to family 22. These CBMs bind xylan but not to cellulose. The gene encoding the mature peptide of these CBMs was fused with an alkaline active GH10 xylanase from Bacillus halodurans S7 and expressed in Escherichia coli. The (His)(6) tagged hybrid protein was purified by immobilized metal affinity chromatography and characterized. Xylan binding by the chimeric protein was influenced by NaCl concentration and pH of the binding medium. Binding increased with increasing salt concentration up to 200 mM. Higher extent of binding was observed under acidic conditions. The fusion of the CBM structures enhanced the hydrolytic efficiency of the xylanase against insoluble xylan, but decreased the stability of the enzyme. The optimum temperature and pH for the activity of the xylanase did not change. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
neapolitana, Thermotoga, chimeric gene, CBM, xylanase, xylan binding domain, Bacillus halodurans
in
Extremophiles
volume
11
issue
1
pages
169 - 177
publisher
Springer
external identifiers
  • wos:000244249100019
  • scopus:33846127494
ISSN
1433-4909
DOI
10.1007/s00792-006-0023-4
language
English
LU publication?
yes
id
d3b68849-2232-42c2-b786-cbae202b223e (old id 674182)
date added to LUP
2007-12-13 15:55:00
date last changed
2017-07-30 03:54:27
@article{d3b68849-2232-42c2-b786-cbae202b223e,
  abstract     = {Xylanase A of Thermotoga neapolitana contains binding domains both at the N- and C-terminal ends of the catalytic domain. In the N-terminal position it contains two carbohydrate-binding modules (CBM) which belong to family 22. These CBMs bind xylan but not to cellulose. The gene encoding the mature peptide of these CBMs was fused with an alkaline active GH10 xylanase from Bacillus halodurans S7 and expressed in Escherichia coli. The (His)(6) tagged hybrid protein was purified by immobilized metal affinity chromatography and characterized. Xylan binding by the chimeric protein was influenced by NaCl concentration and pH of the binding medium. Binding increased with increasing salt concentration up to 200 mM. Higher extent of binding was observed under acidic conditions. The fusion of the CBM structures enhanced the hydrolytic efficiency of the xylanase against insoluble xylan, but decreased the stability of the enzyme. The optimum temperature and pH for the activity of the xylanase did not change.},
  author       = {Mamo, Gashaw and Hatti-Kaul, Rajni and Mattiasson, Bo},
  issn         = {1433-4909},
  keyword      = {neapolitana,Thermotoga,chimeric gene,CBM,xylanase,xylan binding domain,Bacillus halodurans},
  language     = {eng},
  number       = {1},
  pages        = {169--177},
  publisher    = {Springer},
  series       = {Extremophiles},
  title        = {Fusion of carbohydrate binding modules from Thermotoga neapolitana with a family 10 xylanase from Bacillus halodurans S7},
  url          = {http://dx.doi.org/10.1007/s00792-006-0023-4},
  volume       = {11},
  year         = {2007},
}