DB code: S00511

RLCP classification 1.30.36300.2 : Hydrolysis
CATH domain 2.60.120.200 : Jelly Rolls Catalytic domain
E.C. 3.2.1.73
CSA 2ayh
M-CSA 2ayh
MACiE

CATH domain Related DB codes (homologues)
2.60.120.200 : Jelly Rolls S00148 D00535 D00666 M00185 T00064 T00208

Uniprot Enzyme Name
UniprotKB Protein name Synonyms CAZy Pfam
P27051 Beta-glucanase
EC 3.2.1.73
1,3-1,4-beta-D-glucan 4-glucanohydrolase
Endo-beta-1,3-1,4 glucanase
Lichenase
GH16 (Glycoside Hydrolase Family 16)
PF00722 (Glyco_hydro_16)
[Graphical View]
P23904 Beta-glucanase
EC 3.2.1.73
1,3-1,4-beta-D-glucan 4-glucanohydrolase
Endo-beta-1,3-1,4 glucanase
Lichenase
GH16 (Glycoside Hydrolase Family 16)
PF00722 (Glyco_hydro_16)
[Graphical View]

KEGG enzyme name
licheninase
lichenase
beta-(1->4)-D-glucan 4-glucanohydrolase
1,3
1,4-beta-glucan endohydrolase
1,3
1,4-beta-glucan 4-glucanohydrolase
1,3-1,4-beta-D-glucan 4-glucanohydrolase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P23904 GUB_PAEMA Hydrolysis of (1->4)-beta-D-glucosidic linkages in beta-D-glucans containing (1->3)- and (1->4)-bonds.
P27051 GUB_BACLI Hydrolysis of (1->4)-beta-D-glucosidic linkages in beta-D-glucans containing (1->3)- and (1->4)-bonds.

KEGG Pathways
Map code Pathways E.C.

Compound table
Substrates Products Intermediates
KEGG-id C00551 C00478 C00001 C00551
E.C.
Compound beta-D-Glucan Lichenin H2O beta-D-Glucan
Type polysaccharide carbohydrate H2O polysaccharide
ChEBI 15377
PubChem 46173706
439241
22247451
962
46173706
1gbgA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1ajkA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1ajkB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1ajoA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1ajoB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1byhA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Analogue:NBU-GLC-BGC
1cpmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1cpnA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1glhA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1macA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1macB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
2ayhA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1axkA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound
1axkB01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
Swiss-prot;P23904, P45797

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1gbgA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 105;GLU 109
1ajkA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 22;GLU 26
1ajkB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 22;GLU 26
1ajoA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 193;GLU 197
1ajoB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 193;GLU 197
1byhA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 105;GLU 109
1cpmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 47;GLU 51
1cpnA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 47;GLU 51
1glhA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 105;GLU 109
1macA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 103;GLU 107
1macB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 103;GLU 107
2ayhA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 105;GLU 109
1axkA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 47;GLU 51
1axkB01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 47;GLU 51

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[6]
[8]
p.14534
[11]
Fig.8, p.853-854 2
[15]
Scheme 1, p.13846 4
[16]
Scheme 2, p.11340 4
[18]
Scheme 1a 2
[22]
Fig.5, Fig.10, p.369 4
[24]
Scheme 1, p.79 4
[26]
p.2394

References
[1]
Resource
Comments
Medline ID
PubMed ID 1904865
Journal J Biol Chem
Year 1991
Volume 266
Pages 11628-31
Authors Hoj PB, Rodriguez EB, Iser JR, Stick RV, Stone BA
Title Active site-directed inhibition by optically pure epoxyalkyl cellobiosides reveals differences in active site geometry of two 1,3-1,4-beta-D-glucan 4-glucanohydrolases. The importance of epoxide stereochemistry for enzyme inactivation.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 2023245
Journal J Mol Biol
Year 1991
Volume 218
Pages 703-4
Authors Keitel T, Granzin J, Simon O, Borriss R, Thomsen KK, Wessner H, Hohne W, Heinemann U
Title Crystallization of the hybrid Bacillus (1-3,1-4)-beta-glucanase H(A16-M).
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 1354172
Journal FEBS Lett
Year 1992
Volume 308
Pages 141-5
Authors Planas A, Juncosa M, Lloberas J, Querol E
Title Essential catalytic role of Glu134 in endo-beta-1,3-1,4-D-glucan 4-glucanohydrolase from B. licheniformis as determined by site-directed mutagenesis.
Related PDB
Related UniProtKB
[4]
Resource
Comments ACTIVE SITE.
Medline ID 93094208
PubMed ID 1360982
Journal J Biol Chem
Year 1992
Volume 267
Pages 25059-66
Authors Hoj PB, Condron R, Traeger JC, McAuliffe JC, Stone BA
Title Identification of glutamic acid 105 at the active site of Bacillus amyloliquefaciens 1,3-1,4-beta-D-glucan 4-glucanohydrolase using epoxide-based inhibitors.
Related PDB
Related UniProtKB P23904
[5]
Resource
Comments
Medline ID
PubMed ID 8280073
Journal Biochem J
Year 1993
Volume 296
Pages 753-8
Authors Malet C, Jimenez-Barbero J, Bernabe M, Brosa C, Planas A
Title Stereochemical course and structure of the products of the enzymic action of endo-1,3-1,4-beta-D-glucan 4-glucanohydrolase from Bacillus licheniformis.
Related PDB
Related UniProtKB
[6]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID 93281743
PubMed ID 8099449
Journal Proc Natl Acad Sci U S A
Year 1993
Volume 90
Pages 5287-91
Authors Keitel T, Simon O, Borriss R, Heinemann U
Title Molecular and active-site structure of a Bacillus 1,3-1,4-beta-glucanase.
Related PDB 1byh
Related UniProtKB P23904
[7]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 8200344
Journal Eur J Biochem
Year 1994
Volume 222
Pages 203-14
Authors Keitel T, Meldgaard M, Heinemann U
Title Cation binding to a Bacillus (1,3-1,4)-beta-glucanase. Geometry, affinity and effect on protein stability.
Related PDB 1glh
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 8182059
Journal J Biol Chem
Year 1994
Volume 269
Pages 14530-5
Authors Juncosa M, Pons J, Dot T, Querol E, Planas A
Title Identification of active site carboxylic residues in Bacillus licheniformis 1,3-1,4-beta-D-glucan 4-glucanohydrolase by site-directed mutagenesis.
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 8162185
Journal Microbiology
Year 1994
Volume 140
Pages 159-66
Authors Meldgaard M, Svendsen I
Title Different effects of N-glycosylation on the thermostability of highly homologous bacterial (1,3-1,4)-beta-glucanases secreted from yeast.
Related PDB
Related UniProtKB
[10]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 7937966
Journal Proc Natl Acad Sci U S A
Year 1994
Volume 91
Pages 10417-21
Authors Hahn M, Piotukh K, Borriss R, Heinemann U
Title Native-like in vivo folding of a circularly permuted jellyroll protein shown by crystal structure analysis.
Related PDB 1cpm 1cpn
Related UniProtKB
[11]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS).
Medline ID 96028129
PubMed ID 7588726
Journal Eur J Biochem
Year 1995
Volume 232
Pages 849-58
Authors Hahn M, Keitel T, Heinemann U
Title Crystal and molecular structure at 0.16-nm resolution of the hybrid Bacillus endo-1,3-1,4-beta-D-glucan 4-glucanohydrolase H(A16-M).
Related PDB 2ayh
Related UniProtKB P23904
[12]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 7589539
Journal FEBS Lett
Year 1995
Volume 374
Pages 221-4
Authors Hahn M, Pons J, Planas A, Querol E, Heinemann U
Title Crystal structure of Bacillus licheniformis 1,3-1,4-beta-D-glucan 4-glucanohydrolase at 1.8 A resolution.
Related PDB 1gbg
Related UniProtKB
[13]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 7852389
Journal J Biol Chem
Year 1995
Volume 270
Pages 3081-8
Authors Hahn M, Olsen O, Politz O, Borriss R, Heinemann U
Title Crystal structure and site-directed mutagenesis of Bacillus macerans endo-1,3-1,4-beta-glucanase.
Related PDB 1mac
Related UniProtKB
[14]
Resource
Comments
Medline ID
PubMed ID 8746732
Journal Protein Eng
Year 1995
Volume 8
Pages 939-45
Authors Pons J, Planas A, Querol E
Title Contribution of a disulfide bridge to the stability of 1,3-1,4-beta-D-glucan 4-glucanohydrolase from Bacillus licheniformis.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 9374861
Journal Biochemistry
Year 1997
Volume 36
Pages 13838-48
Authors Malet C, Planas A
Title Mechanism of Bacillus 1,3-1,4-beta-D-glucan 4-glucanohydrolases: kinetics and pH studies with 4-methylumbelliferyl beta-D-glucan oligosaccharides.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 9698381
Journal Biochemistry
Year 1998
Volume 37
Pages 11332-42
Authors Viladot JL, de Ramon E, Durany O, Planas A
Title Probing the mechanism of Bacillus 1,3-1,4-beta-D-glucan 4-glucanohydrolases by chemical rescue of inactive mutants at catalytically essential residues.
Related PDB
Related UniProtKB
[17]
Resource
Comments
Medline ID
PubMed ID 9799108
Journal Eur J Biochem
Year 1998
Volume 257
Pages 101-111
Authors Krah M, Misselwitz R, Politz O, Thomsen KK, Welfle H, Borriss R
Title The laminarinase from thermophilic eubacterium Rhodothermus marinus--conformation, stability, and identification of active site carboxylic residues by site-directed mutagenesis.
Related PDB
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 9862456
Journal FEBS Lett
Year 1998
Volume 440
Pages 208-12
Authors Malet C, Planas A
Title From beta-glucanase to beta-glucansynthase: glycosyl transfer to alpha-glycosyl fluorides catalyzed by a mutant endoglucanase lacking its catalytic nucleophile.
Related PDB
Related UniProtKB
[19]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 9618460
Journal Proc Natl Acad Sci U S A
Year 1998
Volume 95
Pages 6613-8
Authors Ay J, Gotz F, Borriss R, Heinemann U
Title Structure and function of the Bacillus hybrid enzyme GluXyn-1: native-like jellyroll fold preserved after insertion of autonomous globular domain.
Related PDB 1axk
Related UniProtKB
[20]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 9489923
Journal Proteins
Year 1998
Volume 30
Pages 155-67
Authors Ay J, Hahn M, Decanniere K, Piotukh K, Borriss R, Heinemann U
Title Crystal structures and properties of de novo circularly permuted 1,3-1,4-beta-glucanases.
Related PDB 1ajk 1ajo
Related UniProtKB
[21]
Resource
Comments
Medline ID
PubMed ID 10587432
Journal Biochemistry
Year 1999
Volume 38
Pages 16092-104
Authors Piotukh K, Serra V, Borriss R, Planas A
Title Protein-carbohydrate interactions defining substrate specificity in Bacillus 1,3-1,4-beta-D-glucan 4-glucanohydrolases as dissected by mutational analysis.
Related PDB
Related UniProtKB
[22]
Resource
Comments
Medline ID
PubMed ID 11150614
Journal Biochim Biophys Acta
Year 2000
Volume 1543
Pages 361-382
Authors Planas A
Title Bacterial 1,3-1,4-beta-glucanases: structure, function and protein engineering.
Related PDB
Related UniProtKB
[23]
Resource
Comments
Medline ID
PubMed ID 11526326
Journal Acta Crystallogr D Biol Crystallogr
Year 2001
Volume 57
Pages 1303-1306
Authors Tsai LC, Shyur LF, Lin SS, Yuan HS
Title Crystallization and preliminary X-ray diffraction analysis of the 1,3-1,4-beta-D-glucanase from Fibrobacter succinogenes.
Related PDB
Related UniProtKB
[24]
Resource
Comments
Medline ID
PubMed ID 11256951
Journal Biochem J
Year 2001
Volume 355
Pages 79-86
Authors Viladot JL, Canals F, Batllori X, Planas A
Title Long-lived glycosyl-enzyme intermediate mimic produced by formate re-activation of a mutant endoglucanase lacking its catalytic nucleophile.
Related PDB
Related UniProtKB
[25]
Resource
Comments
Medline ID
PubMed ID 11279139
Journal J Biol Chem
Year 2001
Volume 276
Pages 17895-17901
Authors Chen JL, Tsai LC, Wen TN, Tang JB, Yuan HS, Shyur LF
Title Directed mutagenesis of apecific active site residues on Fibrobacter succinogenes 1,3-1,4-beta -D-glucanase significantly affects catalysis and enzyme structural stability.
Related PDB
Related UniProtKB
[26]
Resource
Comments
Medline ID
PubMed ID 11841232
Journal Biochemistry
Year 2002
Volume 41
Pages 2388-2395
Authors Ashida H, Maskos K, Li SC, Li YT
Title Characterization of a novel endo-beta-galactosidase specific for releasing the disaccharide GlcNAc alpha 1-->4Gal from glycoconjugates.
Related PDB
Related UniProtKB
[27]
Resource
Comments
Medline ID
PubMed ID 12093295
Journal Biochemistry
Year 2002
Volume 41
Pages 8759-8766
Authors Cheng HL, Tsai LC, Lin SS, Yuan HS, Yang NS, Lee SH, Shyur LF
Title Mutagenesis of Trp(54) and Trp(203) residues on Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase significantly affects catalytic activities of the enzyme.
Related PDB
Related UniProtKB
[28]
Resource
Comments
Medline ID
PubMed ID 12842475
Journal J Mol Biol
Year 2003
Volume 330
Pages 607-20
Authors Tsai LC, Shyur LF, Lee SH, Lin SS, Yuan HS
Title Crystal structure of a natural circularly permuted jellyroll protein: 1,3-1,4-beta-D-glucanase from Fibrobacter succinogenes.
Related PDB
Related UniProtKB

Comments
This enzyme belongs to the glycosidase family-16. This is a retaining glycosidase enzyme, whose configuration of the sugar substrate will be retained after the catalytic reaction.
PDB structures, 1ajk, 1ajo, 1cpm and 2apn, are for the circularly permutated enzyme proteins, whilst that of 1axk is for the artificially fused enzyme with 1,4-beta-xylanase (EC 3.2.1.8; Swiss-prot, P18429).
According to the literature [15], [16] & [22], the catalytic reaction proceeds as follows:
(1) Glycosylation step:Glu105 (of 1byh) makes a nucleophilic attack on the C-1 atom of the polysaccharide substrate, forming a covalent glycosyl-enzyme intermediate through an oxocarbonium-like transition-state. At the same time, Glu109 assists this step as a general acid, to protonate the scissile glycosidic oxygen.
(2) Deglycosylation step:Glu102 acts as a general acid, to activate a water molecule, which then makes a nucleophilic attack on the C-1 atom of the covalent intermediate, resulting in the product release through the same transition state as in the glycosylation step (an oxocarbonium-like transition-state).

Created Updated
2004-07-12 2009-09-30