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 Endo-beta-1,3-1,4 glucanase 1,3-1,4-beta-D-glucan 4-glucanohydrolase Lichenase	GH16 (Glycoside Hydrolase Family 16)	PF00722 (Glyco_hydro_16) [Graphical View]
P23904	Beta-glucanase	EC 3.2.1.73 Endo-beta-1,3-1,4 glucanase 1,3-1,4-beta-D-glucan 4-glucanohydrolase 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
P27051	GUB_BACLI	Hydrolysis of (1->4)-beta-D-glucosidic linkages in beta-D-glucans containing (1->3)- and (1->4)-bonds.
P23904	GUB_PAEMA	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 15377
PubChem	46173706 46173706	439241 439241	22247451 962 22247451 962	46173706 46173706

1gbgA	Unbound	Unbound		Unbound
1ajkA	Unbound	Unbound		Unbound
1ajkB	Unbound	Unbound		Unbound
1ajoA	Unbound	Unbound		Unbound
1ajoB	Unbound	Unbound		Unbound
1byhA	Unbound	Unbound		Analogue:NBU-GLC-BGC
1cpmA	Unbound	Unbound		Unbound
1cpnA	Unbound	Unbound		Unbound
1glhA	Unbound	Unbound		Unbound
1macA	Unbound	Unbound		Unbound
1macB	Unbound	Unbound		Unbound
2ayhA	Unbound	Unbound		Unbound
1axkA01	Unbound	Unbound		Unbound
1axkB01	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	GLU 105;GLU 109
1ajkA	GLU 22;GLU 26
1ajkB	GLU 22;GLU 26
1ajoA	GLU 193;GLU 197
1ajoB	GLU 193;GLU 197
1byhA	GLU 105;GLU 109
1cpmA	GLU 47;GLU 51
1cpnA	GLU 47;GLU 51
1glhA	GLU 105;GLU 109
1macA	GLU 103;GLU 107
1macB	GLU 103;GLU 107
2ayhA	GLU 105;GLU 109
1axkA01	GLU 47;GLU 51
1axkB01	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).

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