Advanced

The structure and characterization of a modular endo-beta-1,4-mannanase from Cellulomonas fimi

Le Nours, K; Anderson, Lars LU ; Stoll, D; Stålbrand, Henrik LU and Lo Leggio, L (2005) In Biochemistry 44(38). p.12700-12708
Abstract
The endo-beta-1,4-mannanase from the soil bacterium Cellulomonas fimi is a modular plant cell wall degrading enzyme involved in the hydrolysis of the backbone of mannan, one of the most abundant polysaccharides of the hemicellulosic network in the plant cell wall. The crystal structure of a recombinant truncated endo-beta-1,4-mannanase from C. fimi (CMan26A-50K) was determined by X-ray crystallography to 2.25 angstrom resolution using the molecular replacement technique. The overall structure of the enzyme consists of a core (beta/alpha)(8)-barrel catalytic module characteristic of clan GH-A, connected via a linker to an immunoglobulin-like module of unknown function. A complex with the oligosaccharide mannotriose to 2.9 angstrom... (More)
The endo-beta-1,4-mannanase from the soil bacterium Cellulomonas fimi is a modular plant cell wall degrading enzyme involved in the hydrolysis of the backbone of mannan, one of the most abundant polysaccharides of the hemicellulosic network in the plant cell wall. The crystal structure of a recombinant truncated endo-beta-1,4-mannanase from C. fimi (CMan26A-50K) was determined by X-ray crystallography to 2.25 angstrom resolution using the molecular replacement technique. The overall structure of the enzyme consists of a core (beta/alpha)(8)-barrel catalytic module characteristic of clan GH-A, connected via a linker to an immunoglobulin-like module of unknown function. A complex with the oligosaccharide mannotriose to 2.9 angstrom resolution has also been obtained. Both the native structure and the complex show a cacodylate ion bound at the -1 subsite, while subsites -2, -3, and -4 are occupied by mannotriose in the complex. Enzyme kinetic analysis and the analysis of hydrolysis products from manno-oligosaccharides and mannopentitol suggest five important active-site cleft subsites. CfMan26A-50K has a high affinity -3 subsite with Phe325 as an aromatic platform, which explains the mannose releasing property of the enzyme. Structural differences with the homologous Cellvibrio japonicus beta-1,4-mannanase (CjMan26A) at the -2 and -3 subsites may explain the poor performance of CfMan26A mutants as "glycosynthases". (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemistry
volume
44
issue
38
pages
12700 - 12708
publisher
The American Chemical Society
external identifiers
  • wos:000232125500008
  • scopus:25444475768
ISSN
0006-2960
DOI
10.1021/bi050779v
language
English
LU publication?
yes
id
498eb105-df24-4e78-990f-6b031c2f5c8c (old id 151427)
date added to LUP
2007-07-06 09:29:23
date last changed
2017-10-22 03:33:56
@article{498eb105-df24-4e78-990f-6b031c2f5c8c,
  abstract     = {The endo-beta-1,4-mannanase from the soil bacterium Cellulomonas fimi is a modular plant cell wall degrading enzyme involved in the hydrolysis of the backbone of mannan, one of the most abundant polysaccharides of the hemicellulosic network in the plant cell wall. The crystal structure of a recombinant truncated endo-beta-1,4-mannanase from C. fimi (CMan26A-50K) was determined by X-ray crystallography to 2.25 angstrom resolution using the molecular replacement technique. The overall structure of the enzyme consists of a core (beta/alpha)(8)-barrel catalytic module characteristic of clan GH-A, connected via a linker to an immunoglobulin-like module of unknown function. A complex with the oligosaccharide mannotriose to 2.9 angstrom resolution has also been obtained. Both the native structure and the complex show a cacodylate ion bound at the -1 subsite, while subsites -2, -3, and -4 are occupied by mannotriose in the complex. Enzyme kinetic analysis and the analysis of hydrolysis products from manno-oligosaccharides and mannopentitol suggest five important active-site cleft subsites. CfMan26A-50K has a high affinity -3 subsite with Phe325 as an aromatic platform, which explains the mannose releasing property of the enzyme. Structural differences with the homologous Cellvibrio japonicus beta-1,4-mannanase (CjMan26A) at the -2 and -3 subsites may explain the poor performance of CfMan26A mutants as "glycosynthases".},
  author       = {Le Nours, K and Anderson, Lars and Stoll, D and Stålbrand, Henrik and Lo Leggio, L},
  issn         = {0006-2960},
  language     = {eng},
  number       = {38},
  pages        = {12700--12708},
  publisher    = {The American Chemical Society},
  series       = {Biochemistry},
  title        = {The structure and characterization of a modular endo-beta-1,4-mannanase from Cellulomonas fimi},
  url          = {http://dx.doi.org/10.1021/bi050779v},
  volume       = {44},
  year         = {2005},
}