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A cellulose-binding module of the Trichoderma reesei @b-mannanase Man5A increases the mannan-hydrolysis of complex substrates

Hägglund, Per LU ; Eriksson, Torny LU ; Collén, Anna LU ; Nerinckx, Wim ; Claeyssens, Marc and Stålbrand, Henrik LU (2003) In Journal of Biotechnology 101(1). p.37-48
Abstract
Endo-@b-1,4-d-mannanases (@b-mannanase; EC 3.2.1.78) are endohydrolases that participate in the degradation of hemicellulose, which is closely associated with cellulose in plant cell walls. The @b-mannanase from Trichoderma reesei (Man5A) is composed of an N-terminal catalytic module and a C-terminal carbohydrate-binding module (CBM). In order to study the properties of the CBM, a construct encoding a mutant of Man5A lacking the part encoding the CBM (Man5A@DCBM), was expressed in T. reesei under the regulation of the Aspergillus nidulans gpdA promoter. The wild-type enzyme was expressed in the same way and both proteins were purified to electrophoretic homogeneity using ion-exchange chromatography. Both enzymes hydrolysed mannopentaose,... (More)
Endo-@b-1,4-d-mannanases (@b-mannanase; EC 3.2.1.78) are endohydrolases that participate in the degradation of hemicellulose, which is closely associated with cellulose in plant cell walls. The @b-mannanase from Trichoderma reesei (Man5A) is composed of an N-terminal catalytic module and a C-terminal carbohydrate-binding module (CBM). In order to study the properties of the CBM, a construct encoding a mutant of Man5A lacking the part encoding the CBM (Man5A@DCBM), was expressed in T. reesei under the regulation of the Aspergillus nidulans gpdA promoter. The wild-type enzyme was expressed in the same way and both proteins were purified to electrophoretic homogeneity using ion-exchange chromatography. Both enzymes hydrolysed mannopentaose, soluble locust bean gum galactomannan and insoluble ivory nut mannan with similar rates. With a mannan/cellulose complex, however, the deletion mutant lacking the CBM showed a significant decrease in hydrolysis. Binding experiments using activity detection of Man5A and Man5A@DCBM suggests that the CBM binds to cellulose but not to mannan. Moreover, the binding of Man5A to cellulose was compared with that of an endoglucanase (Cel7B) from T. reesei. (Less)
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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Carbohydrate-binding module, Hemicellulase, Cellulose, Hemicellulose, Endoglucanase
in
Journal of Biotechnology
volume
101
issue
1
pages
37 - 48
publisher
Elsevier
external identifiers
  • wos:000180677700005
  • pmid:12523968
  • scopus:0346035834
ISSN
1873-4863
DOI
10.1016/S0168-1656(02)00290-0
language
English
LU publication?
yes
id
c1ed5c90-2c13-4ffa-a3a9-d1c09b326e68 (old id 124627)
date added to LUP
2016-04-01 11:39:00
date last changed
2022-02-25 19:18:41
@article{c1ed5c90-2c13-4ffa-a3a9-d1c09b326e68,
  abstract     = {{Endo-@b-1,4-d-mannanases (@b-mannanase; EC 3.2.1.78) are endohydrolases that participate in the degradation of hemicellulose, which is closely associated with cellulose in plant cell walls. The @b-mannanase from Trichoderma reesei (Man5A) is composed of an N-terminal catalytic module and a C-terminal carbohydrate-binding module (CBM). In order to study the properties of the CBM, a construct encoding a mutant of Man5A lacking the part encoding the CBM (Man5A@DCBM), was expressed in T. reesei under the regulation of the Aspergillus nidulans gpdA promoter. The wild-type enzyme was expressed in the same way and both proteins were purified to electrophoretic homogeneity using ion-exchange chromatography. Both enzymes hydrolysed mannopentaose, soluble locust bean gum galactomannan and insoluble ivory nut mannan with similar rates. With a mannan/cellulose complex, however, the deletion mutant lacking the CBM showed a significant decrease in hydrolysis. Binding experiments using activity detection of Man5A and Man5A@DCBM suggests that the CBM binds to cellulose but not to mannan. Moreover, the binding of Man5A to cellulose was compared with that of an endoglucanase (Cel7B) from T. reesei.}},
  author       = {{Hägglund, Per and Eriksson, Torny and Collén, Anna and Nerinckx, Wim and Claeyssens, Marc and Stålbrand, Henrik}},
  issn         = {{1873-4863}},
  keywords     = {{Carbohydrate-binding module; Hemicellulase; Cellulose; Hemicellulose; Endoglucanase}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{37--48}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Biotechnology}},
  title        = {{A cellulose-binding module of the Trichoderma reesei @b-mannanase Man5A increases the mannan-hydrolysis of complex substrates}},
  url          = {{http://dx.doi.org/10.1016/S0168-1656(02)00290-0}},
  doi          = {{10.1016/S0168-1656(02)00290-0}},
  volume       = {{101}},
  year         = {{2003}},
}