EndB, a multidomain family 44 cellulase from Ruminococcus flavefaciens 17, binds to cellulose via a novel cellulose-binding module and to another R-flavefaciens protein via a dockerin domain

M T Rincon, S I McCrae, J Kirby, K P Scott, H J Flint

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

The mechanisms by which cellulolytic enzymes and enzyme complexes in Ruminococcus spp. bind to cellulose are not fully understood. The product of the newly isolated cellulase gene endB from Runtinococcus flavefaciens 17 was purified as a His-tagged product after expression in Escherichia coli and found to be able to bind directly to crystalline cellulose. The ability to bind cellulose is shown to be associated with a novel cellulose-binding module (CBM) located within a region of 200 amino acids that is unrelated to known protein sequences. EndB (808 amino acids) also contains a catalytic domain belonging to glycoside hydrolase family 44 and a C-terminal dockerin-like domain. Purified EndB is also shown to bind specifically via its dockerin domain to a polypeptide of ca. 130 kDa present among supernatant proteins from Avicel-grown R. flavefaciens that attach to cellulose. The protein to which EndB attaches is a strong candidate for the scaffolding component of a cellulosome-like multienzyme complex recently identified in this species (S.-Y. Ding et al., J. Bacteriol. 183:1945-1953, 2001). It is concluded that binding of EndB to cellulose may occur both through its own CBM and potentially also through its involvement in a cellulosome complex.

Original languageEnglish
Pages (from-to)4426-4431
Number of pages6
JournalApplied and Environmental Microbiology
Volume67
Issue number10
Publication statusPublished - Oct 2001

Keywords

  • CELL-SURFACE-STRUCTURES
  • BETA-GLUCANASE GENE
  • ENDO-1,4-BETA-GLUCANASE GENE
  • CLOSTRIDIUM-CELLULOLYTICUM
  • NUCLEOTIDE-SEQUENCE
  • ALBUS
  • ENDOGLUCANASE
  • XYLANASE
  • RUMINOCOCCUS-FLAVEFACIENS-17
  • IDENTIFICATION

Cite this

@article{3205f03d1712457f95db91a670635d70,
title = "EndB, a multidomain family 44 cellulase from Ruminococcus flavefaciens 17, binds to cellulose via a novel cellulose-binding module and to another R-flavefaciens protein via a dockerin domain",
abstract = "The mechanisms by which cellulolytic enzymes and enzyme complexes in Ruminococcus spp. bind to cellulose are not fully understood. The product of the newly isolated cellulase gene endB from Runtinococcus flavefaciens 17 was purified as a His-tagged product after expression in Escherichia coli and found to be able to bind directly to crystalline cellulose. The ability to bind cellulose is shown to be associated with a novel cellulose-binding module (CBM) located within a region of 200 amino acids that is unrelated to known protein sequences. EndB (808 amino acids) also contains a catalytic domain belonging to glycoside hydrolase family 44 and a C-terminal dockerin-like domain. Purified EndB is also shown to bind specifically via its dockerin domain to a polypeptide of ca. 130 kDa present among supernatant proteins from Avicel-grown R. flavefaciens that attach to cellulose. The protein to which EndB attaches is a strong candidate for the scaffolding component of a cellulosome-like multienzyme complex recently identified in this species (S.-Y. Ding et al., J. Bacteriol. 183:1945-1953, 2001). It is concluded that binding of EndB to cellulose may occur both through its own CBM and potentially also through its involvement in a cellulosome complex.",
keywords = "CELL-SURFACE-STRUCTURES, BETA-GLUCANASE GENE, ENDO-1,4-BETA-GLUCANASE GENE, CLOSTRIDIUM-CELLULOLYTICUM, NUCLEOTIDE-SEQUENCE, ALBUS, ENDOGLUCANASE, XYLANASE, RUMINOCOCCUS-FLAVEFACIENS-17, IDENTIFICATION",
author = "Rincon, {M T} and McCrae, {S I} and J Kirby and Scott, {K P} and Flint, {H J}",
year = "2001",
month = "10",
language = "English",
volume = "67",
pages = "4426--4431",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "10",

}

TY - JOUR

T1 - EndB, a multidomain family 44 cellulase from Ruminococcus flavefaciens 17, binds to cellulose via a novel cellulose-binding module and to another R-flavefaciens protein via a dockerin domain

AU - Rincon, M T

AU - McCrae, S I

AU - Kirby, J

AU - Scott, K P

AU - Flint, H J

PY - 2001/10

Y1 - 2001/10

N2 - The mechanisms by which cellulolytic enzymes and enzyme complexes in Ruminococcus spp. bind to cellulose are not fully understood. The product of the newly isolated cellulase gene endB from Runtinococcus flavefaciens 17 was purified as a His-tagged product after expression in Escherichia coli and found to be able to bind directly to crystalline cellulose. The ability to bind cellulose is shown to be associated with a novel cellulose-binding module (CBM) located within a region of 200 amino acids that is unrelated to known protein sequences. EndB (808 amino acids) also contains a catalytic domain belonging to glycoside hydrolase family 44 and a C-terminal dockerin-like domain. Purified EndB is also shown to bind specifically via its dockerin domain to a polypeptide of ca. 130 kDa present among supernatant proteins from Avicel-grown R. flavefaciens that attach to cellulose. The protein to which EndB attaches is a strong candidate for the scaffolding component of a cellulosome-like multienzyme complex recently identified in this species (S.-Y. Ding et al., J. Bacteriol. 183:1945-1953, 2001). It is concluded that binding of EndB to cellulose may occur both through its own CBM and potentially also through its involvement in a cellulosome complex.

AB - The mechanisms by which cellulolytic enzymes and enzyme complexes in Ruminococcus spp. bind to cellulose are not fully understood. The product of the newly isolated cellulase gene endB from Runtinococcus flavefaciens 17 was purified as a His-tagged product after expression in Escherichia coli and found to be able to bind directly to crystalline cellulose. The ability to bind cellulose is shown to be associated with a novel cellulose-binding module (CBM) located within a region of 200 amino acids that is unrelated to known protein sequences. EndB (808 amino acids) also contains a catalytic domain belonging to glycoside hydrolase family 44 and a C-terminal dockerin-like domain. Purified EndB is also shown to bind specifically via its dockerin domain to a polypeptide of ca. 130 kDa present among supernatant proteins from Avicel-grown R. flavefaciens that attach to cellulose. The protein to which EndB attaches is a strong candidate for the scaffolding component of a cellulosome-like multienzyme complex recently identified in this species (S.-Y. Ding et al., J. Bacteriol. 183:1945-1953, 2001). It is concluded that binding of EndB to cellulose may occur both through its own CBM and potentially also through its involvement in a cellulosome complex.

KW - CELL-SURFACE-STRUCTURES

KW - BETA-GLUCANASE GENE

KW - ENDO-1,4-BETA-GLUCANASE GENE

KW - CLOSTRIDIUM-CELLULOLYTICUM

KW - NUCLEOTIDE-SEQUENCE

KW - ALBUS

KW - ENDOGLUCANASE

KW - XYLANASE

KW - RUMINOCOCCUS-FLAVEFACIENS-17

KW - IDENTIFICATION

M3 - Article

VL - 67

SP - 4426

EP - 4431

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 10

ER -