EzCatDB: D00500

DB codeD00500
RLCP classification1.30.4920.981
CATH domainDomain 11.50.10.10Catalytic domain
Domain 22.60.40.10
E.C.3.2.1.3
CSA1agm

CATH domainRelated DB codes (homologues)
1.50.10.10S00531,S00048,S00845,D00167,M00192,T00245,T00246
2.60.40.10M00131,T00257,T00005,M00113,M00127,M00132,M00323,M00325,M00327,M00329,M00330,M00331,M00332,T00307,D00166,M00112,M00193,T00063,T00065,T00067,T00245

Enzyme Name
Swiss-protKEGG

P69327P69328
Protein nameGlucoamylaseGlucoamylaseglucan 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
SynonymsEC 3.2.1.3
Glucan 1,4-alpha-glucosidase
1,4-alpha-D-glucan glucohydrolase
EC 3.2.1.3
Glucan 1,4-alpha-glucosidase
1,4-alpha-D-glucan glucohydrolase

KEGG pathways
MAP codePathways
MAP00500Starch and sucrose metabolism

Swiss-prot:Accession NumberP69327P69328
Entry nameAMYG_ASPAWAMYG_ASPNG
ActivityHydrolysis of terminal (1->4)-linked alpha-D- glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose.Hydrolysis of terminal (1->4)-linked alpha-D- glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose.
Subunit

Subcellular location

Cofactor



SubstratesProductsintermediates
KEGG-idC00718C00001C00718C00221
CompoundAmyloseH2OAmylosebeta-D-Glucose
TypepolysaccharideH2Opolysaccharidecarbohydrate
1agmAAnalogue:ACR
UnboundUnboundUnbound
1dogAUnbound
Analogue:NOJ 481UnboundTransition-state-analogue:NOJ 480
1gahAAnalogue:ACR
UnboundUnboundUnbound
1gaiAAnalogue:GAC
UnboundUnboundUnbound
1glmAUnbound
UnboundUnboundUnbound
1glyAUnbound
UnboundUnboundUnbound
3glyAUnbound
UnboundUnboundUnbound
1ac0AUnbound
UnboundUnboundUnbound
1aczAUnbound
UnboundUnboundUnbound
1kulAUnbound
UnboundUnboundUnbound
1kumAUnbound
UnboundUnboundUnbound

Active-site residues
resource
PDB;1agm & literature [9], [14]
pdbCatalytic residues
1agmATYR 48;ARG 54;GLU 179;GLU 400
1dogATYR 48;ARG 54;GLU 179;GLU 400
1gahATYR 48;ARG 54;GLU 179;GLU 400
1gaiATYR 48;ARG 54;GLU 179;GLU 400
1glmATYR 48;ARG 54;GLU 179;GLU 400
1glyATYR 48;ARG 54;GLU 179;GLU 400
3glyATYR 48;ARG 54;GLU 179;GLU 400
1ac0A
1aczA
1kulA
1kumA

References for Catalytic Mechanism
ReferencesSectionsNo. 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]
CommentsMUTAGENESIS OF TRP-144.
PubMed ID2510150
JournalProtein Eng
Year1989
Volume2
Pages621-5
AuthorsSierks MR, Ford C, Reilly PJ, Svensson B
TitleSite-directed mutagenesis at the active site Trp120 of Aspergillus awamori glucoamylase.
Related Swiss-protP69327
[2]
PubMed ID2108020
JournalEur J Biochem
Year1990
Volume188
Pages29-38
AuthorsSvensson B, Clarke AJ, Svendsen I, Moller H
TitleIdentification of carboxylic acid residues in glucoamylase G2 from Aspergillus niger that participate in catalysis and substrate binding.
[3]
CommentsACTIVE SITES, AND MUTAGENESIS.
PubMed ID1970434
JournalProtein Eng
Year1990
Volume3
Pages193-8
AuthorsSierks MR, Ford C, Reilly PJ, Svensson B
TitleCatalytic mechanism of fungal glucoamylase as defined by mutagenesis of Asp176, Glu179 and Glu180 in the enzyme from Aspergillus awamori.
Related Swiss-protP69327
[4]
CommentsCONFORMATION OF O-GLYCOSYLATED REGION.
PubMed ID1546955
JournalBiochem J
Year1992
Volume282
Pages423-8
AuthorsWilliamson G, Belshaw NJ, Williamson MP
TitleO-glycosylation in Aspergillus glucoamylase. Conformation and role in binding.
Related Swiss-protP69328
[5]
PubMed ID1390684
JournalBiochemistry
Year1992
Volume31
Pages8972-7
AuthorsSierks MR, Bock K, Refn S, Svensson B
TitleActive site similarities of glucose dehydrogenase, glucose oxidase, and glucoamylase probed by deoxygenated substrates.
[6]
PubMed ID1633817
JournalEur J Biochem
Year1992
Volume207
Pages661-70
AuthorsWilliamson G, Belshaw NJ, Noel TR, Ring SG, Williamson MP
TitleO-glycosylation and stability. Unfolding of glucoamylase induced by heat and guanidine hydrochloride.
[7]
CommentsX-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, N-GLYCOSYLATION, O-GLYCOSYLATION, AND DISULFIDE BONDS.
Medline ID92406872
PubMed ID1527049
JournalJ Biol Chem
Year1992
Volume267
Pages19291-8
AuthorsAleshin A, Golubev A, Firsov LM, Honzatko RB
TitleCrystal structure of glucoamylase from Aspergillus awamori var. X100 to 2.2-A resolution.
Related PDB1gly
Related Swiss-protP69327
[8]
CommentsCHARACTERIZATION OF CATALYTIC DOMAIN.
Medline ID93277459
PubMed ID8503847
JournalBiochem J
Year1993
Volume292
Pages197-202
AuthorsStoffer B, Frandsen TP, Busk PK, Schneider P, Svendsen I, Svensson B
TitleProduction, purification and characterization of the catalytic domain of glucoamylase from Aspergillus niger.
Related Swiss-protP69328
[9]
CommentsX-ray crystallography
PubMed ID8431441
JournalBiochemistry
Year1993
Volume32
Pages1618-26
AuthorsHarris EM, Aleshin AE, Firsov LM, Honzatko RB
TitleRefined structure for the complex of 1-deoxynojirimycin with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution.
Related PDB1dog
[10]
PubMed ID8373772
JournalBiochemistry
Year1993
Volume32
Pages9686-93
AuthorsOlsen K, Christensen U, Sierks MR, Svensson B
TitleReaction mechanisms of Trp120-->Phe and wild-type glucoamylases from Aspergillus niger. Interactions with maltooligodextrins and acarbose.
[11]
PubMed ID8424940
JournalBiochemistry
Year1993
Volume32
Pages1113-7
AuthorsSierks MR, Svensson B
TitleFunctional roles of the invariant aspartic acid 55, tyrosine 306, and aspartic acid 309 in glucoamylase from Aspergillus awamori studied by mutagenesis.
[12]
PubMed ID8309943
JournalProtein Eng
Year1993
Volume6
Pages939-46
AuthorsBakir U, Coutinho PM, Sullivan PA, Ford C, Reilly PJ
TitleCassette mutagenesis of Aspergillus awamori glucoamylase near its general acid residue to probe its catalytic and pH properties.
[13]
CommentsMUTAGENESIS.
PubMed ID8433972
JournalProtein Eng
Year1993
Volume6
Pages75-9
AuthorsSierks MR, Ford C, Reilly PJ, Svensson B
TitleFunctional roles and subsite locations of Leu177, Trp178 and Asn182 of Aspergillus awamori glucoamylase determined by site-directed mutagenesis.
Related Swiss-protP69327
[14]
PubMed ID7947792
JournalBiochemistry
Year1994
Volume33
Pages13808-16
AuthorsFrandsen TP, Dupont C, Lehmbeck J, Stoffer B, Sierks MR, Honzatko RB, Svensson B
TitleSite-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.
[15]
PubMed ID8033904
JournalEur J Biochem
Year1994
Volume223
Pages293-302
AuthorsFirsov LM, Neustroev KN, Aleshin AE, Metzler CM, Metzler DE, Scott RD, Stoffer B, Christensen T, Svensson B
TitleNMR spectroscopy of exchangeable protons of glucoamylase and of complexes with inhibitors in the 9-15-ppm range.
[16]
CommentsX-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 25-495.
Medline ID94253149
PubMed ID8195212
JournalJ Biol Chem
Year1994
Volume269
Pages15631-9
AuthorsAleshin AE, Firsov LM, Honzatko RB
TitleRefined structure for the complex of acarbose with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution.
Related PDB1agm
Related Swiss-protP69327
[17]
CommentsX-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, N-GLYCOSYLATION, AND O-GLYCOSYLATION.
Medline ID94231577
PubMed ID8176747
JournalJ Mol Biol
Year1994
Volume238
Pages575-91
AuthorsAleshin AE, Hoffman C, Firsov LM, Honzatko RB
TitleRefined crystal structures of glucoamylase from Aspergillus awamori var. X100.
Related PDB1glm,3gly
Related Swiss-protP69327
[18]
PubMed ID7937705
JournalProtein Eng
Year1994
Volume7
Pages749-60
AuthorsCoutinho PM, Reilly PJ
TitleStructural similarities in glucoamylase by hydrophobic cluster analysis.
[19]
PubMed ID8177888
JournalProtein Eng
Year1994
Volume7
Pages393-400
AuthorsCoutinho PM, Reilly PJ
TitleStructure-function relationships in the catalytic and starch binding domains of glucoamylase.
[20]
PubMed ID7809026
JournalProtein Eng
Year1994
Volume7
Pages1005-12
AuthorsFlory N, Gorman M, Coutinho PM, Ford C, Reilly PJ
TitleThermosensitive mutants of Aspergillus awamori glucoamylase by random mutagenesis: inactivation kinetics and structural interpretation.
[21]
PubMed ID7640270
JournalBiochemistry
Year1995
Volume34
Pages10162-9
AuthorsFrandsen TP, Christensen T, Stoffer B, Lehmbeck J, Dupont C, Honzatko RB, Svensson B
TitleMutational 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.
[22]
PubMed ID7765970
JournalBiosci Biotechnol Biochem
Year1995
Volume59
Pages16-20
AuthorsGoto M, Kuwano E, Kanlayakrit W, Hayashida S
TitleRole of the carbohydrate moiety of a glucoamylase from Aspergillus awamori var. kawachi in the digestion of raw starch.
[23]
PubMed ID7588803
JournalEur J Biochem
Year1995
Volume233
Pages568-78
AuthorsJacks AJ, Sorimachi K, Le Gal-Coeffet MF, Williamson G, Archer DB, Williamson MP
Title1H and 15N assignments and secondary structure of the starch-binding domain of glucoamylase from Aspergillus niger.
[24]
PubMed ID7821430
JournalFEBS Lett
Year1995
Volume358
Pages57-61
AuthorsStoffer B, Aleshin AE, Firsov LM, Svensson B, Honzatko RB
TitleRefined 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.
[25]
PubMed ID8586614
JournalJ Biochem (Tokyo)
Year1995
Volume117
Pages1024-8
AuthorsTanaka A, Fukada H, Takahashi K
TitleDifferential scanning calorimetric studies on the domain structure of Aspergillus glucoamylase.
[26]
PubMed ID8535779
JournalStructure
Year1995
Volume3
Pages853-9
AuthorsDavies G, Henrissat B
TitleStructures and mechanisms of glycosyl hydrolases.
[27]
CommentsX-ray crystallography
PubMed ID8679589
JournalBiochemistry
Year1996
Volume35
Pages8319-28
AuthorsAleshin AE, Stoffer B, Firsov LM, Svensson B, Honzatko RB
TitleCrystallographic complexes of glucoamylase with maltooligosaccharide analogs: relationship of stereochemical distortions at the nonreducing end to the catalytic mechanism.
Related PDB1gah,1gai
[28]
PubMed ID8679632
JournalBiochemistry
Year1996
Volume35
Pages8696-704
AuthorsFierobe HP, Stoffer BB, Frandsen TP, Svensson B
TitleMutational 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.
[29]
PubMed ID8608145
JournalBiochemistry
Year1996
Volume35
Pages3050-8
AuthorsNatarajan S, Sierks MR
TitleFunctional and structural roles of the highly conserved Trp120 loop region of glucoamylase from Aspergillus awamori.
[30]
PubMed ID8639668
JournalBiochemistry
Year1996
Volume35
Pages1865-71
AuthorsSierks MR, Svensson B
TitleCatalytic mechanism of glucoamylase probed by mutagenesis in conjunction with hydrolysis of alpha-D-glucopyranosyl fluoride and maltooligosaccharides.
[31]
PubMed ID8672288
JournalJ Biotechnol
Year1996
Volume46
Pages85-93
AuthorsMacKenzie DA, Spencer JA, Le Gal-Coeffet MF, Archer DB
TitleEfficient production from Aspergillus niger of a heterologous protein and an individual protein domain, heavy isotope-labelled, for structure-function analysis.
[32]
PubMed ID8890914
JournalJ Mol Biol
Year1996
Volume263
Pages79-89
AuthorsFrandsen TP, Stoffer BB, Palcic MM, Hof S, Svensson B
TitleStructure and energetics of the glucoamylase-isomaltose transition-state complex probed by using modeling and deoxygenated substrates coupled with site-directed mutagenesis.
[33]
CommentsSTRUCTURE BY NMR OF 533-640.
Medline ID96266494
PubMed ID8683599
JournalJ Mol Biol
Year1996
Volume259
Pages970-87
AuthorsSorimachi K, Jacks AJ, Le Gal-Coeffet MF, Williamson G, Archer DB, Williamson MP
TitleSolution structure of the granular starch binding domain of glucoamylase from Aspergillus niger by nuclear magnetic resonance spectroscopy.
Related PDB1kul,1kum
Related Swiss-protP69328
[34]
PubMed ID8862550
JournalProtein Eng
Year1996
Volume9
Pages499-505
AuthorsChen HM, Li Y, Panda T, Buehler FU, Ford C, Reilly PJ
TitleEffect of replacing helical glycine residues with alanines on reversible and irreversible stability and production of Aspergillus awamori glucoamylase.
[35]
PubMed ID9200704
JournalBiochemistry
Year1997
Volume36
Pages7535-9
AuthorsWilliamson MP, Le Gal-Coeffet MF, Sorimachi K, Furniss CS, Archer DB, Williamson G
TitleFunction of conserved tryptophans in the Aspergillus niger glucoamylase 1 starch binding domain.
[36]
PubMed ID9461285
JournalEur J Biochem
Year1997
Volume250
Pages638-45
AuthorsChristensen T, Stoffer BB, Svensson B, Christensen U
TitleSome details of the reaction mechanism of glucoamylase from Aspergillus niger--kinetic and structural studies on Trp52-->Phe and Trp317-->Phe mutants.
[37]
PubMed ID9488144
JournalProtein Eng
Year1997
Volume10
Pages1199-204
AuthorsLi Y, Reilly PJ, Ford C
TitleEffect of introducing proline residues on the stability of Aspergillus awamori.
[38]
PubMed ID9051738
JournalProtein Eng
Year1997
Volume10
Pages81-7
AuthorsStoffer BB, Dupont C, Frandsen TP, Lehmbeck J, Svensson B
TitleGlucoamylase mutants in the conserved active-site segment Trp170-Tyr175 located at a distance from the site of catalysis.
[39]
PubMed ID9061788
JournalProteins
Year1997
Volume27
Pages235-48
AuthorsCoutinho PM, Dowd MK, Reilly PJ
TitleAutomated docking of monosaccharide substrates and analogues and methyl alpha-acarviosinide in the glucoamylase active site.
[40]
PubMed ID9188734
JournalProteins
Year1997
Volume28
Pages162-73
AuthorsCoutinho PM, Dowd MK, Reilly PJ
TitleAutomated docking of glucosyl disaccharides in the glucoamylase active site.
[41]
PubMed ID9365988
JournalProteins
Year1997
Volume29
Pages334-47
AuthorsCoutinho PM, Reilly PJ
TitleGlucoamylase structural, functional, and evolutionary relationships.
[42]
CommentsSTRUCTURE BY NMR OF 533-640.
Medline ID97341228
PubMed ID9195884
JournalStructure
Year1997
Volume5
Pages647-61
AuthorsSorimachi K, Le Gal-Coeffet MF, Williamson G, Archer DB, Williamson MP
TitleSolution structure of the granular starch binding domain of Aspergillus niger glucoamylase bound to beta-cyclodextrin.
Related PDB1acz,1ac0
Related Swiss-protP69328
[43]
PubMed ID9649747
JournalBiochem Soc Trans
Year1998
Volume26
Pages198-204
AuthorsGottschalk 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
TitleStructure, function and protein engineering of starch-degrading enzymes.
[44]
PubMed ID9521694
JournalBiochemistry
Year1998
Volume37
Pages3753-9
AuthorsFierobe HP, Clarke AJ, Tull D, Svensson B
TitleEnzymatic properties of the cysteinesulfinic acid derivative of the catalytic-base mutant Glu400-->Cys of glucoamylase from Aspergillus awamori.
[45]
PubMed ID9521693
JournalBiochemistry
Year1998
Volume37
Pages3743-52
AuthorsFierobe HP, Mirgorodskaya E, McGuire KA, Roepstorff P, Svensson B, Clarke AJ
TitleRestoration of catalytic activity beyond wild-type level in glucoamylase from Aspergillus awamori by oxidation of the Glu400-->Cys catalytic-base mutant to cysteinesulfinic acid.
[46]
PubMed ID9671514
JournalBiochemistry
Year1998
Volume37
Pages10446-52
AuthorsSigurskjold BW, Christensen T, Payre N, Cottaz S, Driguez H, Svensson B
TitleThermodynamics 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.
[47]
PubMed ID9972233
JournalBiosci Biotechnol Biochem
Year1998
Volume62
Pages2127-32
AuthorsTanaka A, Karita S, Kosuge Y, Senoo K, Obata H, Kitamoto N
TitleThermal unfolding of the starch binding domain of Aspergillus niger glucoamylase.
[48]
PubMed ID9605546
JournalProtein Eng
Year1998
Volume11
Pages119-26
AuthorsFang TY, Coutinho PM, Reilly PJ, Ford C
TitleMutations to alter Aspergillus awamori glucoamylase selectivity. I. Tyr48Phe49-->Trp, Tyr116-->Trp, Tyr175-->Phe, Arg241-->Lys, Ser411-->Ala and Ser411-->Gly.
[49]
PubMed ID9681871
JournalProtein Eng
Year1998
Volume11
Pages383-8
AuthorsFang TY, Ford C
TitleProtein engineering of Aspergillus awamori glucoamylase to increase its pH optimum.
[50]
PubMed ID9605547
JournalProtein Eng
Year1998
Volume11
Pages127-33
AuthorsFang TY, Honzatko RB, Reilly PJ, Ford C
TitleMutations to alter Aspergillus awamori glucoamylase selectivity. II. Mutation of residues 119 and 121.
[51]
PubMed ID9749918
JournalProtein Eng
Year1998
Volume11
Pages661-7
AuthorsLi Y, Coutinho PM, Ford C
TitleEffect on thermostability and catalytic activity of introducing disulfide bonds into Aspergillus awamori glucoamylase.
[52]
PubMed ID10320360
JournalBiochemistry
Year1999
Volume38
Pages6300-10
AuthorsChristensen T, Svensson B, Sigurskjold BW
TitleThermodynamics of reversible and irreversible unfolding and domain interactions of glucoamylase from Aspergillus niger studied by differential scanning and isothermal titration calorimetry.
[53]
PubMed ID10540741
JournalBiosci Biotechnol Biochem
Year1999
Volume63
Pages1548-52
AuthorsTanaka A, Ohya M, Yamamoto T, Nakagawa C, Tsuji T, Senoo K, Obata H
TitleSteady-state inhibitory kinetic studies on the ligand binding modes of Aspergillus niger glucoamylase.
[54]
PubMed ID10449316
JournalCurr Opin Biotechnol
Year1999
Volume10
Pages353-7
AuthorsFord C
TitleImproving operating performance of glucoamylase by mutagenesis.
[55]
PubMed ID10452542
JournalFEBS Lett
Year1999
Volume456
Pages119-25
AuthorsJanecek S, Sevcik J
TitleThe evolution of starch-binding domain.
[56]
PubMed ID10913265
JournalBiochemistry
Year2000
Volume39
Pages8585-92
AuthorsSierks MR, Svensson B
TitleEnergetic and mechanistic studies of glucoamylase using molecular recognition of maltose OH groups coupled with site-directed mutagenesis.
[57]
PubMed ID10630989
JournalBiochemistry
Year2000
Volume39
Pages300-6
AuthorsWeimar T, Stoffer B, Svensson B, Pinto BM
TitleComplexes of glucoamylase with maltoside heteroanalogues: bound ligand conformations by use of transferred NOE experiments and molecular modeling.
[58]
PubMed ID11150611
JournalBiochim Biophys Acta
Year2000
Volume1543
Pages275-93
AuthorsSauer J, Sigurskjold BW, Christensen U, Frandsen TP, Mirgorodskaya E, Harrison M, Roepstorff P, Svensson B
TitleGlucoamylase: structure/function relationships, and protein engineering.
[59]
PubMed ID11054460
JournalProtein Eng
Year2000
Volume13
Pages655-9
AuthorsLiu HL, Doleyres Y, Coutinho PM, Ford C, Reilly PJ
TitleReplacement and deletion mutations in the catalytic domain and belt region of Aspergillus awamori glucoamylase to enhance thermostability.
[60]
PubMed ID11478902
JournalBiochemistry
Year2001
Volume40
Pages9336-46
AuthorsSauer J, Christensen T, Frandsen TP, Mirgorodskaya E, McGuire KA, Driguez H, Roepstorff P, Sigurskjold BW, Svensson B
TitleStability and function of interdomain linker variants of glucoamylase 1 from Aspergillus niger.
[61]
PubMed ID11508823
JournalGen Physiol Biophys
Year2001
Volume20
Pages7-32
AuthorsHorvathova V, Janecek S, Sturdik E
TitleAmylolytic enzymes: molecular aspects of their properties.
[62]
PubMed ID11700070
JournalJ Mol Biol
Year2001
Volume313
Pages1149-59
AuthorsGiardina T, Gunning AP, Juge N, Faulds CB, Furniss CS, Svensson B, Morris VJ, Williamson G
TitleBoth binding sites of the starch-binding domain of Aspergillus niger glucoamylase are essential for inducing a conformational change in amylose.
[63]
PubMed ID12445478
JournalBiochim Biophys Acta
Year2002
Volume1601
Pages163-71
AuthorsChristensen T, Frandsen TP, Kaarsholm NC, Svensson B, Sigurskjold BW
TitlePhysicochemical characterisation of the two active site mutants Trp(52)-->Phe and Asp(55)-->Val of glucoamylase from Aspergillus niger.
[64]
PubMed ID12529155
JournalJ Biomol Struct Dyn
Year2003
Volume20
Pages567-74
AuthorsLiu HL, Wang WC, Hsu CM
TitleMolecular dynamics simulations of the unfolding mechanism of the catalytic domain from Aspergillus awamori var. X100 glucoamylase.
[65]
PubMed ID12646689
JournalProtein Eng
Year2003
Volume16
Pages19-25
AuthorsLiu HL, Wang WC
TitleProtein engineering to improve the thermostability of glucoamylase from Aspergillus awamori based on molecular dynamics simulations.
[66]
PubMed ID15963591
JournalJ Biotechnol
Year2005
Volume118
Pages167-76
AuthorsLatorre-Garcia L, Adam AC, Manzanares P, Polaina J
TitleImproving the amylolytic activity of Saccharomyces cerevisiae glucoamylase by the addition of a starch binding domain.

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.
----------

createdupdated
2004-07-092009-02-26


Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2010)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)

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