DB code: D00500

RLCP classification 1.30.4920.981 : Hydrolysis
CATH domain 1.50.10.10 : Glycosyltransferase Catalytic domain
2.60.40.10 : Immunoglobulin-like
E.C. 3.2.1.3
CSA 1agm
M-CSA 1agm
MACiE

CATH domain Related DB codes (homologues)
2.60.40.10 : Immunoglobulin-like M00131 T00257 T00005 M00113 M00127 M00132 M00323 M00325 M00327 M00329 M00330 M00331 M00332 T00307 D00166 M00112 M00193 T00063 T00065 T00067 T00245
1.50.10.10 : Glycosyltransferase S00531 S00048 S00845 D00167 M00192 T00245 T00246

Uniprot Enzyme Name
UniprotKB Protein name Synonyms Pfam CAZy
P69327 Glucoamylase
EC 3.2.1.3
1,4-alpha-D-glucan glucohydrolase
Glucan 1,4-alpha-glucosidase
PF00686 (CBM_20)
PF00723 (Glyco_hydro_15)
[Graphical View]
P69328 Glucoamylase
EC 3.2.1.3
1,4-alpha-D-glucan glucohydrolase
Glucan 1,4-alpha-glucosidase
PF00686 (CBM_20)
PF00723 (Glyco_hydro_15)
[Graphical View]
CBM20 (Carbohydrate-Binding Module Family 20)
GH15 (Glycoside Hydrolase Family 15)

KEGG enzyme name
glucan 1,4-alpha-glucosidase
glucoamylase
amyloglucosidase
gamma-amylase
lysosomal alpha-glucosidase
acid maltase
exo-1,4-alpha-glucosidase
glucose amylase
gamma-1,4-glucan glucohydrolase
acid maltase
1,4-alpha-D-glucan glucohydrolase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P69328 AMYG_ASPNG Hydrolysis of terminal (1->4)-linked alpha-D- glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose.
P69327 AMYG_ASPAW Hydrolysis of terminal (1->4)-linked alpha-D- glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose.

KEGG Pathways
Map code Pathways E.C.
MAP00500 Starch and sucrose metabolism

Compound table
Substrates Products Intermediates
KEGG-id C00718 C00001 C00718 C00221
E.C.
Compound Amylose H2O Amylose beta-D-Glucose
Type polysaccharide H2O polysaccharide carbohydrate
ChEBI 15377
15903
PubChem 22247451
962
64689
1agmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:ACR Unbound Unbound Unbound
1dogA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Analogue:NOJ_481 Unbound Transition-state-analogue:NOJ_480
1gahA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:ACR Unbound Unbound Unbound
1gaiA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:GAC Unbound Unbound Unbound
1glmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound
1glyA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound
3glyA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound
1ac0A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound
1aczA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound
1kulA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound
1kumA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
PDB;1agm & literature [9], [14]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1agmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
1dogA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
1gahA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
1gaiA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
1glmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
1glyA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
3glyA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain TYR 48;ARG 54;GLU 179;GLU 400
1ac0A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain
1aczA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain
1kulA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain
1kumA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[2]
SCHEME 2, p.35-37
[3]
p.194-197
[9]
p.1623-1625
[14]
Fig.3, p.13812-13814
[16]
p.15637-15638
[21]
p.10166-10168
[26]
Fig.1, p.853
[27]
[30]
p.1869-1870
[43]
Fig.1
[45]
Fig.1
[58]
Fig.5, p.279-280

References
[1]
Resource
Comments MUTAGENESIS OF TRP-144.
Medline ID
PubMed ID 2510150
Journal Protein Eng
Year 1989
Volume 2
Pages 621-5
Authors Sierks MR, Ford C, Reilly PJ, Svensson B
Title Site-directed mutagenesis at the active site Trp120 of Aspergillus awamori glucoamylase.
Related PDB
Related UniProtKB P69327
[2]
Resource
Comments
Medline ID
PubMed ID 2108020
Journal Eur J Biochem
Year 1990
Volume 188
Pages 29-38
Authors Svensson B, Clarke AJ, Svendsen I, Moller H
Title Identification of carboxylic acid residues in glucoamylase G2 from Aspergillus niger that participate in catalysis and substrate binding.
Related PDB
Related UniProtKB
[3]
Resource
Comments ACTIVE SITES, AND MUTAGENESIS.
Medline ID
PubMed ID 1970434
Journal Protein Eng
Year 1990
Volume 3
Pages 193-8
Authors Sierks MR, Ford C, Reilly PJ, Svensson B
Title Catalytic mechanism of fungal glucoamylase as defined by mutagenesis of Asp176, Glu179 and Glu180 in the enzyme from Aspergillus awamori.
Related PDB
Related UniProtKB P69327
[4]
Resource
Comments CONFORMATION OF O-GLYCOSYLATED REGION.
Medline ID
PubMed ID 1546955
Journal Biochem J
Year 1992
Volume 282
Pages 423-8
Authors Williamson G, Belshaw NJ, Williamson MP
Title O-glycosylation in Aspergillus glucoamylase. Conformation and role in binding.
Related PDB
Related UniProtKB P69328
[5]
Resource
Comments
Medline ID
PubMed ID 1390684
Journal Biochemistry
Year 1992
Volume 31
Pages 8972-7
Authors Sierks MR, Bock K, Refn S, Svensson B
Title Active site similarities of glucose dehydrogenase, glucose oxidase, and glucoamylase probed by deoxygenated substrates.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 1633817
Journal Eur J Biochem
Year 1992
Volume 207
Pages 661-70
Authors Williamson G, Belshaw NJ, Noel TR, Ring SG, Williamson MP
Title O-glycosylation and stability. Unfolding of glucoamylase induced by heat and guanidine hydrochloride.
Related PDB
Related UniProtKB
[7]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, N-GLYCOSYLATION, O-GLYCOSYLATION, AND DISULFIDE BONDS.
Medline ID 92406872
PubMed ID 1527049
Journal J Biol Chem
Year 1992
Volume 267
Pages 19291-8
Authors Aleshin A, Golubev A, Firsov LM, Honzatko RB
Title Crystal structure of glucoamylase from Aspergillus awamori var. X100 to 2.2-A resolution.
Related PDB 1gly
Related UniProtKB P69327
[8]
Resource
Comments CHARACTERIZATION OF CATALYTIC DOMAIN.
Medline ID 93277459
PubMed ID 8503847
Journal Biochem J
Year 1993
Volume 292
Pages 197-202
Authors Stoffer B, Frandsen TP, Busk PK, Schneider P, Svendsen I, Svensson B
Title Production, purification and characterization of the catalytic domain of glucoamylase from Aspergillus niger.
Related PDB
Related UniProtKB P69328
[9]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 8431441
Journal Biochemistry
Year 1993
Volume 32
Pages 1618-26
Authors Harris EM, Aleshin AE, Firsov LM, Honzatko RB
Title Refined structure for the complex of 1-deoxynojirimycin with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution.
Related PDB 1dog
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 8373772
Journal Biochemistry
Year 1993
Volume 32
Pages 9686-93
Authors Olsen K, Christensen U, Sierks MR, Svensson B
Title Reaction mechanisms of Trp120-->Phe and wild-type glucoamylases from Aspergillus niger. Interactions with maltooligodextrins and acarbose.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 8424940
Journal Biochemistry
Year 1993
Volume 32
Pages 1113-7
Authors Sierks MR, Svensson B
Title Functional roles of the invariant aspartic acid 55, tyrosine 306, and aspartic acid 309 in glucoamylase from Aspergillus awamori studied by mutagenesis.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 8309943
Journal Protein Eng
Year 1993
Volume 6
Pages 939-46
Authors Bakir U, Coutinho PM, Sullivan PA, Ford C, Reilly PJ
Title Cassette mutagenesis of Aspergillus awamori glucoamylase near its general acid residue to probe its catalytic and pH properties.
Related PDB
Related UniProtKB
[13]
Resource
Comments MUTAGENESIS.
Medline ID
PubMed ID 8433972
Journal Protein Eng
Year 1993
Volume 6
Pages 75-9
Authors Sierks MR, Ford C, Reilly PJ, Svensson B
Title Functional roles and subsite locations of Leu177, Trp178 and Asn182 of Aspergillus awamori glucoamylase determined by site-directed mutagenesis.
Related PDB
Related UniProtKB P69327
[14]
Resource
Comments
Medline ID
PubMed ID 7947792
Journal Biochemistry
Year 1994
Volume 33
Pages 13808-16
Authors Frandsen TP, Dupont C, Lehmbeck J, Stoffer B, Sierks MR, Honzatko RB, Svensson B
Title Site-directed mutagenesis of the catalytic base glutamic acid 400 in glucoamylase from Aspergillus niger and of tyrosine 48 and glutamine 401, both hydrogen-bonded to the gamma-carboxylate group of glutamic acid 400.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 8033904
Journal Eur J Biochem
Year 1994
Volume 223
Pages 293-302
Authors Firsov LM, Neustroev KN, Aleshin AE, Metzler CM, Metzler DE, Scott RD, Stoffer B, Christensen T, Svensson B
Title NMR spectroscopy of exchangeable protons of glucoamylase and of complexes with inhibitors in the 9-15-ppm range.
Related PDB
Related UniProtKB
[16]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 25-495.
Medline ID 94253149
PubMed ID 8195212
Journal J Biol Chem
Year 1994
Volume 269
Pages 15631-9
Authors Aleshin AE, Firsov LM, Honzatko RB
Title Refined structure for the complex of acarbose with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution.
Related PDB 1agm
Related UniProtKB P69327
[17]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, N-GLYCOSYLATION, AND O-GLYCOSYLATION.
Medline ID 94231577
PubMed ID 8176747
Journal J Mol Biol
Year 1994
Volume 238
Pages 575-91
Authors Aleshin AE, Hoffman C, Firsov LM, Honzatko RB
Title Refined crystal structures of glucoamylase from Aspergillus awamori var. X100.
Related PDB 1glm 3gly
Related UniProtKB P69327
[18]
Resource
Comments
Medline ID
PubMed ID 7937705
Journal Protein Eng
Year 1994
Volume 7
Pages 749-60
Authors Coutinho PM, Reilly PJ
Title Structural similarities in glucoamylase by hydrophobic cluster analysis.
Related PDB
Related UniProtKB
[19]
Resource
Comments
Medline ID
PubMed ID 8177888
Journal Protein Eng
Year 1994
Volume 7
Pages 393-400
Authors Coutinho PM, Reilly PJ
Title Structure-function relationships in the catalytic and starch binding domains of glucoamylase.
Related PDB
Related UniProtKB
[20]
Resource
Comments
Medline ID
PubMed ID 7809026
Journal Protein Eng
Year 1994
Volume 7
Pages 1005-12
Authors Flory N, Gorman M, Coutinho PM, Ford C, Reilly PJ
Title Thermosensitive mutants of Aspergillus awamori glucoamylase by random mutagenesis: inactivation kinetics and structural interpretation.
Related PDB
Related UniProtKB
[21]
Resource
Comments
Medline ID
PubMed ID 7640270
Journal Biochemistry
Year 1995
Volume 34
Pages 10162-9
Authors Frandsen TP, Christensen T, Stoffer B, Lehmbeck J, Dupont C, Honzatko RB, Svensson B
Title Mutational analysis of the roles in catalysis and substrate recognition of arginines 54 and 305, aspartic acid 309, and tryptophan 317 located at subsites 1 and 2 in glucoamylase from Aspergillus niger.
Related PDB
Related UniProtKB
[22]
Resource
Comments
Medline ID
PubMed ID 7765970
Journal Biosci Biotechnol Biochem
Year 1995
Volume 59
Pages 16-20
Authors Goto M, Kuwano E, Kanlayakrit W, Hayashida S
Title Role of the carbohydrate moiety of a glucoamylase from Aspergillus awamori var. kawachi in the digestion of raw starch.
Related PDB
Related UniProtKB
[23]
Resource
Comments
Medline ID
PubMed ID 7588803
Journal Eur J Biochem
Year 1995
Volume 233
Pages 568-78
Authors Jacks AJ, Sorimachi K, Le Gal-Coeffet MF, Williamson G, Archer DB, Williamson MP
Title 1H and 15N assignments and secondary structure of the starch-binding domain of glucoamylase from Aspergillus niger.
Related PDB
Related UniProtKB
[24]
Resource
Comments
Medline ID
PubMed ID 7821430
Journal FEBS Lett
Year 1995
Volume 358
Pages 57-61
Authors Stoffer B, Aleshin AE, Firsov LM, Svensson B, Honzatko RB
Title Refined structure for the complex of D-gluco-dihydroacarbose with glucoamylase from Aspergillus awamori var. X100 to 2.2 A resolution: dual conformations for extended inhibitors bound to the active site of glucoamylase.
Related PDB
Related UniProtKB
[25]
Resource
Comments
Medline ID
PubMed ID 8586614
Journal J Biochem (Tokyo)
Year 1995
Volume 117
Pages 1024-8
Authors Tanaka A, Fukada H, Takahashi K
Title Differential scanning calorimetric studies on the domain structure of Aspergillus glucoamylase.
Related PDB
Related UniProtKB
[26]
Resource
Comments
Medline ID
PubMed ID 8535779
Journal Structure
Year 1995
Volume 3
Pages 853-9
Authors Davies G, Henrissat B
Title Structures and mechanisms of glycosyl hydrolases.
Related PDB
Related UniProtKB
[27]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 8679589
Journal Biochemistry
Year 1996
Volume 35
Pages 8319-28
Authors Aleshin AE, Stoffer B, Firsov LM, Svensson B, Honzatko RB
Title Crystallographic complexes of glucoamylase with maltooligosaccharide analogs: relationship of stereochemical distortions at the nonreducing end to the catalytic mechanism.
Related PDB 1gah 1gai
Related UniProtKB
[28]
Resource
Comments
Medline ID
PubMed ID 8679632
Journal Biochemistry
Year 1996
Volume 35
Pages 8696-704
Authors Fierobe HP, Stoffer BB, Frandsen TP, Svensson B
Title Mutational modulation of substrate bond-type specificity and thermostability of glucoamylase from Aspergillus awamori by replacement with short homologue active site sequences and thiol/disulfide engineering.
Related PDB
Related UniProtKB
[29]
Resource
Comments
Medline ID
PubMed ID 8608145
Journal Biochemistry
Year 1996
Volume 35
Pages 3050-8
Authors Natarajan S, Sierks MR
Title Functional and structural roles of the highly conserved Trp120 loop region of glucoamylase from Aspergillus awamori.
Related PDB
Related UniProtKB
[30]
Resource
Comments
Medline ID
PubMed ID 8639668
Journal Biochemistry
Year 1996
Volume 35
Pages 1865-71
Authors Sierks MR, Svensson B
Title Catalytic mechanism of glucoamylase probed by mutagenesis in conjunction with hydrolysis of alpha-D-glucopyranosyl fluoride and maltooligosaccharides.
Related PDB
Related UniProtKB
[31]
Resource
Comments
Medline ID
PubMed ID 8672288
Journal J Biotechnol
Year 1996
Volume 46
Pages 85-93
Authors MacKenzie DA, Spencer JA, Le Gal-Coeffet MF, Archer DB
Title Efficient production from Aspergillus niger of a heterologous protein and an individual protein domain, heavy isotope-labelled, for structure-function analysis.
Related PDB
Related UniProtKB
[32]
Resource
Comments
Medline ID
PubMed ID 8890914
Journal J Mol Biol
Year 1996
Volume 263
Pages 79-89
Authors Frandsen TP, Stoffer BB, Palcic MM, Hof S, Svensson B
Title Structure and energetics of the glucoamylase-isomaltose transition-state complex probed by using modeling and deoxygenated substrates coupled with site-directed mutagenesis.
Related PDB
Related UniProtKB
[33]
Resource
Comments STRUCTURE BY NMR OF 533-640.
Medline ID 96266494
PubMed ID 8683599
Journal J Mol Biol
Year 1996
Volume 259
Pages 970-87
Authors Sorimachi K, Jacks AJ, Le Gal-Coeffet MF, Williamson G, Archer DB, Williamson MP
Title Solution structure of the granular starch binding domain of glucoamylase from Aspergillus niger by nuclear magnetic resonance spectroscopy.
Related PDB 1kul 1kum
Related UniProtKB P69328
[34]
Resource
Comments
Medline ID
PubMed ID 8862550
Journal Protein Eng
Year 1996
Volume 9
Pages 499-505
Authors Chen HM, Li Y, Panda T, Buehler FU, Ford C, Reilly PJ
Title Effect of replacing helical glycine residues with alanines on reversible and irreversible stability and production of Aspergillus awamori glucoamylase.
Related PDB
Related UniProtKB
[35]
Resource
Comments
Medline ID
PubMed ID 9200704
Journal Biochemistry
Year 1997
Volume 36
Pages 7535-9
Authors Williamson MP, Le Gal-Coeffet MF, Sorimachi K, Furniss CS, Archer DB, Williamson G
Title Function of conserved tryptophans in the Aspergillus niger glucoamylase 1 starch binding domain.
Related PDB
Related UniProtKB
[36]
Resource
Comments
Medline ID
PubMed ID 9461285
Journal Eur J Biochem
Year 1997
Volume 250
Pages 638-45
Authors Christensen T, Stoffer BB, Svensson B, Christensen U
Title Some details of the reaction mechanism of glucoamylase from Aspergillus niger--kinetic and structural studies on Trp52-->Phe and Trp317-->Phe mutants.
Related PDB
Related UniProtKB
[37]
Resource
Comments
Medline ID
PubMed ID 9488144
Journal Protein Eng
Year 1997
Volume 10
Pages 1199-204
Authors Li Y, Reilly PJ, Ford C
Title Effect of introducing proline residues on the stability of Aspergillus awamori.
Related PDB
Related UniProtKB
[38]
Resource
Comments
Medline ID
PubMed ID 9051738
Journal Protein Eng
Year 1997
Volume 10
Pages 81-7
Authors Stoffer BB, Dupont C, Frandsen TP, Lehmbeck J, Svensson B
Title Glucoamylase mutants in the conserved active-site segment Trp170-Tyr175 located at a distance from the site of catalysis.
Related PDB
Related UniProtKB
[39]
Resource
Comments
Medline ID
PubMed ID 9061788
Journal Proteins
Year 1997
Volume 27
Pages 235-48
Authors Coutinho PM, Dowd MK, Reilly PJ
Title Automated docking of monosaccharide substrates and analogues and methyl alpha-acarviosinide in the glucoamylase active site.
Related PDB
Related UniProtKB
[40]
Resource
Comments
Medline ID
PubMed ID 9188734
Journal Proteins
Year 1997
Volume 28
Pages 162-73
Authors Coutinho PM, Dowd MK, Reilly PJ
Title Automated docking of glucosyl disaccharides in the glucoamylase active site.
Related PDB
Related UniProtKB
[41]
Resource
Comments
Medline ID
PubMed ID 9365988
Journal Proteins
Year 1997
Volume 29
Pages 334-47
Authors Coutinho PM, Reilly PJ
Title Glucoamylase structural, functional, and evolutionary relationships.
Related PDB
Related UniProtKB
[42]
Resource
Comments STRUCTURE BY NMR OF 533-640.
Medline ID 97341228
PubMed ID 9195884
Journal Structure
Year 1997
Volume 5
Pages 647-61
Authors Sorimachi K, Le Gal-Coeffet MF, Williamson G, Archer DB, Williamson MP
Title Solution structure of the granular starch binding domain of Aspergillus niger glucoamylase bound to beta-cyclodextrin.
Related PDB 1acz 1ac0
Related UniProtKB P69328
[43]
Resource
Comments
Medline ID
PubMed ID 9649747
Journal Biochem Soc Trans
Year 1998
Volume 26
Pages 198-204
Authors Gottschalk TE, Fierobe HP, Mirgorodskaya E, Clarke AJ, Tull D, Sigurskjold BW, Christensen T, Payre N, Frandsen TP, Juge N, McGuire KA, Cottaz S, Roepstorff P, Driguez H, Williamson G, Svensson B
Title Structure, function and protein engineering of starch-degrading enzymes.
Related PDB
Related UniProtKB
[44]
Resource
Comments
Medline ID
PubMed ID 9521694
Journal Biochemistry
Year 1998
Volume 37
Pages 3753-9
Authors Fierobe HP, Clarke AJ, Tull D, Svensson B
Title Enzymatic properties of the cysteinesulfinic acid derivative of the catalytic-base mutant Glu400-->Cys of glucoamylase from Aspergillus awamori.
Related PDB
Related UniProtKB
[45]
Resource
Comments
Medline ID
PubMed ID 9521693
Journal Biochemistry
Year 1998
Volume 37
Pages 3743-52
Authors Fierobe HP, Mirgorodskaya E, McGuire KA, Roepstorff P, Svensson B, Clarke AJ
Title Restoration of catalytic activity beyond wild-type level in glucoamylase from Aspergillus awamori by oxidation of the Glu400-->Cys catalytic-base mutant to cysteinesulfinic acid.
Related PDB
Related UniProtKB
[46]
Resource
Comments
Medline ID
PubMed ID 9671514
Journal Biochemistry
Year 1998
Volume 37
Pages 10446-52
Authors Sigurskjold BW, Christensen T, Payre N, Cottaz S, Driguez H, Svensson B
Title Thermodynamics of binding of heterobidentate ligands consisting of spacer-connected acarbose and beta-cyclodextrin to the catalytic and starch-binding domains of glucoamylase from Aspergillus niger shows that the catalytic and starch-binding sites are in close proximity in space.
Related PDB
Related UniProtKB
[47]
Resource
Comments
Medline ID
PubMed ID 9972233
Journal Biosci Biotechnol Biochem
Year 1998
Volume 62
Pages 2127-32
Authors Tanaka A, Karita S, Kosuge Y, Senoo K, Obata H, Kitamoto N
Title Thermal unfolding of the starch binding domain of Aspergillus niger glucoamylase.
Related PDB
Related UniProtKB
[48]
Resource
Comments
Medline ID
PubMed ID 9605546
Journal Protein Eng
Year 1998
Volume 11
Pages 119-26
Authors Fang TY, Coutinho PM, Reilly PJ, Ford C
Title Mutations to alter Aspergillus awamori glucoamylase selectivity. I. Tyr48Phe49-->Trp, Tyr116-->Trp, Tyr175-->Phe, Arg241-->Lys, Ser411-->Ala and Ser411-->Gly.
Related PDB
Related UniProtKB
[49]
Resource
Comments
Medline ID
PubMed ID 9681871
Journal Protein Eng
Year 1998
Volume 11
Pages 383-8
Authors Fang TY, Ford C
Title Protein engineering of Aspergillus awamori glucoamylase to increase its pH optimum.
Related PDB
Related UniProtKB
[50]
Resource
Comments
Medline ID
PubMed ID 9605547
Journal Protein Eng
Year 1998
Volume 11
Pages 127-33
Authors Fang TY, Honzatko RB, Reilly PJ, Ford C
Title Mutations to alter Aspergillus awamori glucoamylase selectivity. II. Mutation of residues 119 and 121.
Related PDB
Related UniProtKB
[51]
Resource
Comments
Medline ID
PubMed ID 9749918
Journal Protein Eng
Year 1998
Volume 11
Pages 661-7
Authors Li Y, Coutinho PM, Ford C
Title Effect on thermostability and catalytic activity of introducing disulfide bonds into Aspergillus awamori glucoamylase.
Related PDB
Related UniProtKB
[52]
Resource
Comments
Medline ID
PubMed ID 10320360
Journal Biochemistry
Year 1999
Volume 38
Pages 6300-10
Authors Christensen T, Svensson B, Sigurskjold BW
Title Thermodynamics of reversible and irreversible unfolding and domain interactions of glucoamylase from Aspergillus niger studied by differential scanning and isothermal titration calorimetry.
Related PDB
Related UniProtKB
[53]
Resource
Comments
Medline ID
PubMed ID 10540741
Journal Biosci Biotechnol Biochem
Year 1999
Volume 63
Pages 1548-52
Authors Tanaka A, Ohya M, Yamamoto T, Nakagawa C, Tsuji T, Senoo K, Obata H
Title Steady-state inhibitory kinetic studies on the ligand binding modes of Aspergillus niger glucoamylase.
Related PDB
Related UniProtKB
[54]
Resource
Comments
Medline ID
PubMed ID 10449316
Journal Curr Opin Biotechnol
Year 1999
Volume 10
Pages 353-7
Authors Ford C
Title Improving operating performance of glucoamylase by mutagenesis.
Related PDB
Related UniProtKB
[55]
Resource
Comments
Medline ID
PubMed ID 10452542
Journal FEBS Lett
Year 1999
Volume 456
Pages 119-25
Authors Janecek S, Sevcik J
Title The evolution of starch-binding domain.
Related PDB
Related UniProtKB
[56]
Resource
Comments
Medline ID
PubMed ID 10913265
Journal Biochemistry
Year 2000
Volume 39
Pages 8585-92
Authors Sierks MR, Svensson B
Title Energetic and mechanistic studies of glucoamylase using molecular recognition of maltose OH groups coupled with site-directed mutagenesis.
Related PDB
Related UniProtKB
[57]
Resource
Comments
Medline ID
PubMed ID 10630989
Journal Biochemistry
Year 2000
Volume 39
Pages 300-6
Authors Weimar T, Stoffer B, Svensson B, Pinto BM
Title Complexes of glucoamylase with maltoside heteroanalogues: bound ligand conformations by use of transferred NOE experiments and molecular modeling.
Related PDB
Related UniProtKB
[58]
Resource
Comments
Medline ID
PubMed ID 11150611
Journal Biochim Biophys Acta
Year 2000
Volume 1543
Pages 275-93
Authors Sauer J, Sigurskjold BW, Christensen U, Frandsen TP, Mirgorodskaya E, Harrison M, Roepstorff P, Svensson B
Title Glucoamylase: structure/function relationships, and protein engineering.
Related PDB
Related UniProtKB
[59]
Resource
Comments
Medline ID
PubMed ID 11054460
Journal Protein Eng
Year 2000
Volume 13
Pages 655-9
Authors Liu HL, Doleyres Y, Coutinho PM, Ford C, Reilly PJ
Title Replacement and deletion mutations in the catalytic domain and belt region of Aspergillus awamori glucoamylase to enhance thermostability.
Related PDB
Related UniProtKB
[60]
Resource
Comments
Medline ID
PubMed ID 11478902
Journal Biochemistry
Year 2001
Volume 40
Pages 9336-46
Authors Sauer J, Christensen T, Frandsen TP, Mirgorodskaya E, McGuire KA, Driguez H, Roepstorff P, Sigurskjold BW, Svensson B
Title Stability and function of interdomain linker variants of glucoamylase 1 from Aspergillus niger.
Related PDB
Related UniProtKB
[61]
Resource
Comments
Medline ID
PubMed ID 11508823
Journal Gen Physiol Biophys
Year 2001
Volume 20
Pages 7-32
Authors Horvathova V, Janecek S, Sturdik E
Title Amylolytic enzymes: molecular aspects of their properties.
Related PDB
Related UniProtKB
[62]
Resource
Comments
Medline ID
PubMed ID 11700070
Journal J Mol Biol
Year 2001
Volume 313
Pages 1149-59
Authors Giardina T, Gunning AP, Juge N, Faulds CB, Furniss CS, Svensson B, Morris VJ, Williamson G
Title Both binding sites of the starch-binding domain of Aspergillus niger glucoamylase are essential for inducing a conformational change in amylose.
Related PDB
Related UniProtKB
[63]
Resource
Comments
Medline ID
PubMed ID 12445478
Journal Biochim Biophys Acta
Year 2002
Volume 1601
Pages 163-71
Authors Christensen T, Frandsen TP, Kaarsholm NC, Svensson B, Sigurskjold BW
Title Physicochemical characterisation of the two active site mutants Trp(52)-->Phe and Asp(55)-->Val of glucoamylase from Aspergillus niger.
Related PDB
Related UniProtKB
[64]
Resource
Comments
Medline ID
PubMed ID 12529155
Journal J Biomol Struct Dyn
Year 2003
Volume 20
Pages 567-74
Authors Liu HL, Wang WC, Hsu CM
Title Molecular dynamics simulations of the unfolding mechanism of the catalytic domain from Aspergillus awamori var. X100 glucoamylase.
Related PDB
Related UniProtKB
[65]
Resource
Comments
Medline ID
PubMed ID 12646689
Journal Protein Eng
Year 2003
Volume 16
Pages 19-25
Authors Liu HL, Wang WC
Title Protein engineering to improve the thermostability of glucoamylase from Aspergillus awamori based on molecular dynamics simulations.
Related PDB
Related UniProtKB
[66]
Resource
Comments
Medline ID
PubMed ID 15963591
Journal J Biotechnol
Year 2005
Volume 118
Pages 167-76
Authors Latorre-Garcia L, Adam AC, Manzanares P, Polaina J
Title Improving the amylolytic activity of Saccharomyces cerevisiae glucoamylase by the addition of a starch binding domain.
Related PDB
Related UniProtKB

Comments
This enzyme belongs to glycosidase family-15, which has an inverting mechanism.
According to the literature [9], [14], [21], [27] & [30], the catalytic reaction proceeds as follows:
(1) Glu179 acts as a general acid, to protonate the glycosidic oxygen of the scissile bond, leading to a formation of an oxocarbenium ion transition-state.
(2) Glu179, whose sidechain is now deprotonated, Glu400, Tyr48 probably stabilize the transition-state. Arg54 might modulate the activity of Glu179.
(3) Tyr48 might modulate the pKa of Glu400.
(4) Glu400 acts as a general base, to activate a catalytic water.
(5) The activated water makes a nucelophilic attack on the oxocarbenium ion, to complete the reaction.
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Created Updated
2004-07-09 2009-02-26