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Protein

Amylosucrase

Gene

ams

Organism
Neisseria polysaccharea
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the synthesis of alpha-glucan from sucrose. Catalyzes, in addition, sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis, these two sucrose isomers being obtained by glucosyl transfer onto fructose.1 Publication

Catalytic activityi

Sucrose + ((1->4)-alpha-D-glucosyl)(n) = D-fructose + ((1->4)-alpha-D-glucosyl)(n+1).2 Publications

Enzyme regulationi

Amylosucrase favors hydrolysis at low sucrose concentrations, and polymerization at high sucrose concentrations. Competitively inhibited by fructose.

Kineticsi

This enzyme does not present a classic Michaelis-Menten behavior for sucrose consumption, that could be related to the presence of a second sucrose binding site. Nevertheless, it is possible to model sucrose consumption rate versus sucrose concentration by two different Michaelis-Menten equations whose apparent kinetic constants are given just above.

  1. KM=1.9 mM for sucrose (in the sucrose consumption assay, when initial sucrose < 20 mM)1 Publication
  2. KM=1.7 mM for sucrose (in the sucrose hydrolysis reaction, when initial sucrose < 20 mM)1 Publication
  3. KM=1.9 mM for sucrose (in the polymerization reaction, when initial sucrose < 20 mM)1 Publication
  4. KM=50.2 mM for sucrose (in the sucrose consumption assay, when initial sucrose > 20 mM)1 Publication
  5. KM=38.7 mM for sucrose (in the sucrose hydrolysis reaction, when initial sucrose > 20 mM)1 Publication
  1. Vmax=470 µmol/min/g enzyme for sucrose consumption (when initial sucrose < 20 mM)1 Publication
  2. Vmax=288 µmol/min/g enzyme for sucrose hydrolysis (when initial sucrose < 20 mM)1 Publication
  3. Vmax=147 µmol/min/g enzyme for polymerization reaction (when initial sucrose < 20 mM)1 Publication
  4. Vmax=1100 µmol/min/g enzyme for sucrose consumption (when initial sucrose > 20 mM)1 Publication
  5. Vmax=472 µmol/min/g enzyme for sucrose hydrolysis (when initial sucrose > 20 mM)1 Publication
  6. Vmax=1620 µmol/min/g enzyme for polymerization reaction (when initial sucrose > 20 mM)1 Publication

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei152 – 1521Substrate3 Publications
Binding sitei195 – 1951Substrate3 Publications
Binding sitei262 – 2621Substrate3 Publications
Binding sitei292 – 2921Substrate3 Publications
Active sitei294 – 2941Nucleophile1 Publication1 Publication
Active sitei336 – 3361Proton donor1 Publication
Binding sitei400 – 4001Substrate3 Publications
Binding sitei401 – 4011Substrate3 Publications
Sitei452 – 4521Transition state stabilizerBy similarity
Binding sitei517 – 5171Substrate3 Publications

GO - Molecular functioni

GO - Biological processi

Complete GO annotation...

Keywords - Molecular functioni

Glycosyltransferase, Transferase

Enzyme and pathway databases

BRENDAi2.4.1.4. 3596.
SABIO-RKQ9ZEU2.

Protein family/group databases

CAZyiGH13. Glycoside Hydrolase Family 13.

Names & Taxonomyi

Protein namesi
Recommended name:
Amylosucrase (EC:2.4.1.42 Publications)
Gene namesi
Name:ams
OrganismiNeisseria polysaccharea
Taxonomic identifieri489 [NCBI]
Taxonomic lineageiBacteriaProteobacteriaBetaproteobacteriaNeisserialesNeisseriaceaeNeisseria

Subcellular locationi

  • Secreted 1 Publication

GO - Cellular componenti

Complete GO annotation...

Keywords - Cellular componenti

Secreted

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi195 – 1951H → Q: 98% reduction of activity. 1 Publication
Mutagenesisi294 – 2941D → E or N: Complete loss of activity. 1 Publication
Mutagenesisi308 – 3081E → Q: No effect. 1 Publication
Mutagenesisi336 – 3361E → Q: Complete loss of activity. 1 Publication
Mutagenesisi352 – 3521E → Q: 30% reduction of activity. 1 Publication
Mutagenesisi400 – 4001H → N: 97% reduction of activity. 1 Publication
Mutagenesisi401 – 4011D → E: Almost complete loss of activity. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 636636AmylosucrasePRO_0000045154Add
BLAST

Interactioni

Subunit structurei

Monomer.4 Publications

Protein-protein interaction databases

STRINGi546267.NEIPOLOT_01155.

Structurei

Secondary structure

1
636
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Helixi13 – 197Combined sources
Helixi20 – 223Combined sources
Helixi25 – 339Combined sources
Helixi35 – 5925Combined sources
Helixi65 – 8218Combined sources
Helixi85 – 9511Combined sources
Helixi98 – 1025Combined sources
Beta strandi108 – 1114Combined sources
Helixi113 – 1175Combined sources
Helixi120 – 1256Combined sources
Helixi127 – 1337Combined sources
Beta strandi136 – 1405Combined sources
Turni152 – 1554Combined sources
Beta strandi160 – 1634Combined sources
Turni165 – 1673Combined sources
Helixi170 – 18213Combined sources
Beta strandi186 – 1916Combined sources
Beta strandi194 – 1974Combined sources
Helixi201 – 2077Combined sources
Helixi211 – 2133Combined sources
Beta strandi219 – 2235Combined sources
Helixi224 – 2296Combined sources
Turni230 – 2323Combined sources
Turni238 – 2403Combined sources
Beta strandi245 – 2473Combined sources
Beta strandi253 – 2553Combined sources
Beta strandi257 – 2593Combined sources
Beta strandi262 – 2654Combined sources
Helixi270 – 28415Combined sources
Turni285 – 2873Combined sources
Beta strandi289 – 2935Combined sources
Helixi296 – 2983Combined sources
Beta strandi307 – 3093Combined sources
Helixi311 – 32717Combined sources
Beta strandi332 – 3354Combined sources
Helixi341 – 3444Combined sources
Helixi345 – 3473Combined sources
Beta strandi352 – 3576Combined sources
Helixi359 – 37113Combined sources
Helixi375 – 3839Combined sources
Beta strandi392 – 3976Combined sources
Helixi409 – 4146Combined sources
Helixi419 – 43012Combined sources
Beta strandi442 – 4443Combined sources
Turni448 – 4503Combined sources
Beta strandi454 – 4563Combined sources
Helixi459 – 4635Combined sources
Helixi465 – 4673Combined sources
Helixi472 – 48514Combined sources
Beta strandi486 – 4938Combined sources
Helixi496 – 4983Combined sources
Helixi506 – 5083Combined sources
Turni510 – 5145Combined sources
Helixi516 – 5205Combined sources
Helixi526 – 5294Combined sources
Turni530 – 5334Combined sources
Helixi538 – 55518Combined sources
Helixi557 – 5593Combined sources
Beta strandi564 – 5663Combined sources
Beta strandi574 – 5796Combined sources
Turni580 – 5823Combined sources
Beta strandi583 – 5886Combined sources
Beta strandi590 – 5923Combined sources
Beta strandi594 – 5963Combined sources
Turni598 – 6036Combined sources
Beta strandi606 – 6105Combined sources
Turni611 – 6133Combined sources
Beta strandi616 – 6183Combined sources
Beta strandi623 – 6253Combined sources
Beta strandi630 – 6345Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1G5AX-ray1.40A13-636[»]
1JG9X-ray1.66A13-636[»]
1JGIX-ray2.00A13-636[»]
1MVYX-ray2.00A13-636[»]
1MW0X-ray2.01A13-636[»]
1MW1X-ray2.10A13-636[»]
1MW2X-ray2.10A13-636[»]
1MW3X-ray2.00A13-636[»]
1S46X-ray2.20A13-636[»]
1ZS2X-ray2.16A13-636[»]
3UEQX-ray1.85A13-636[»]
4FLOX-ray2.20A13-636[»]
4FLQX-ray2.50A13-636[»]
4FLRX-ray2.40A13-636[»]
4FLSX-ray2.30A13-636[»]
ProteinModelPortaliQ9ZEU2.
SMRiQ9ZEU2. Positions 13-636.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiQ9ZEU2.

Family & Domainsi

Sequence similaritiesi

Belongs to the glycosyl hydrolase 13 family.Curated

Phylogenomic databases

eggNOGiENOG4107QW3. Bacteria.
COG0366. LUCA.

Family and domain databases

Gene3Di2.60.40.1180. 1 hit.
3.20.20.80. 2 hits.
InterProiIPR015902. Glyco_hydro_13.
IPR006047. Glyco_hydro_13_cat_dom.
IPR013780. Glyco_hydro_b.
IPR013781. Glyco_hydro_catalytic_dom.
IPR017853. Glycoside_hydrolase_SF.
[Graphical view]
PANTHERiPTHR10357. PTHR10357. 2 hits.
PfamiPF00128. Alpha-amylase. 1 hit.
[Graphical view]
SMARTiSM00642. Aamy. 1 hit.
[Graphical view]
SUPFAMiSSF51445. SSF51445. 1 hit.

Sequencei

Sequence statusi: Complete.

Q9ZEU2-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MLTPTQQVGL ILQYLKTRIL DIYTPEQRAG IEKSEDWRQF SRRMDTHFPK
60 70 80 90 100
LMNELDSVYG NNEALLPMLE MLLAQAWQSY SQRNSSLKDI DIARENNPDW
110 120 130 140 150
ILSNKQVGGV CYVDLFAGDL KGLKDKIPYF QELGLTYLHL MPLFKCPEGK
160 170 180 190 200
SDGGYAVSSY RDVNPALGTI GDLREVIAAL HEAGISAVVD FIFNHTSNEH
210 220 230 240 250
EWAQRCAAGD PLFDNFYYIF PDRRMPDQYD RTLREIFPDQ HPGGFSQLED
260 270 280 290 300
GRWVWTTFNS FQWDLNYSNP WVFRAMAGEM LFLANLGVDI LRMDAVAFIW
310 320 330 340 350
KQMGTSCENL PQAHALIRAF NAVMRIAAPA VFFKSEAIVH PDQVVQYIGQ
360 370 380 390 400
DECQIGYNPL QMALLWNTLA TREVNLLHQA LTYRHNLPEH TAWVNYVRSH
410 420 430 440 450
DDIGWTFADE DAAYLGISGY DHRQFLNRFF VNRFDGSFAR GVPFQYNPST
460 470 480 490 500
GDCRVSGTAA ALVGLAQDDP HAVDRIKLLY SIALSTGGLP LIYLGDEVGT
510 520 530 540 550
LNDDDWSQDS NKSDDSRWAH RPRYNEALYA QRNDPSTAAG QIYQGLRHMI
560 570 580 590 600
AVRQSNPRFD GGRLVTFNTN NKHIIGYIRN NALLAFGNFS EYPQTVTAHT
610 620 630
LQAMPFKAHD LIGGKTVSLN QDLTLQPYQV MWLEIA
Length:636
Mass (Da):72,344
Last modified:May 1, 1999 - v1
Checksum:iB7656C19BF1A2065
GO

Sequence cautioni

The sequence described in PubMed:9150231 differs from that shown. Reason: Frameshift at positions 183, 203, 274, 313, 507, 538 and 579. Curated

Experimental Info

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Sequence conflicti14 – 141Y → H in AAM51153 (PubMed:12517860).Curated
Sequence conflicti84 – 852NS → SA in AAM51153 (PubMed:12517860).Curated
Sequence conflicti89 – 891D → N in AAM51153 (PubMed:12517860).Curated
Sequence conflicti93 – 931A → E in AAM51153 (PubMed:12517860).Curated
Sequence conflicti159 – 1591S → T AA sequence (PubMed:9150231).Curated
Sequence conflicti171 – 1711G → D in AAM51153 (PubMed:12517860).Curated
Sequence conflicti208 – 2081Missing AA sequence (PubMed:9150231).Curated
Sequence conflicti392 – 3921A → S in AAM51153 (PubMed:12517860).Curated
Sequence conflicti437 – 4371S → T AA sequence (PubMed:9150231).Curated
Sequence conflicti449 – 4491S → N in AAM51153 (PubMed:12517860).Curated
Sequence conflicti459 – 4591A → S in AAM51153 (PubMed:12517860).Curated
Sequence conflicti463 – 4631V → A in AAM51153 (PubMed:12517860).Curated
Sequence conflicti468 – 4681D → N in AAM51153 (PubMed:12517860).Curated
Sequence conflicti474 – 4741D → S in AAM51153 (PubMed:12517860).Curated
Sequence conflicti501 – 5011L → P in AAM51153 (PubMed:12517860).Curated
Sequence conflicti505 – 5106DWSQDS → GWAQDG in AAM51153 (PubMed:12517860).Curated

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AJ011781 Genomic DNA. Translation: CAA09772.1.
AY099335 Genomic DNA. Translation: AAM51153.1.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AJ011781 Genomic DNA. Translation: CAA09772.1.
AY099335 Genomic DNA. Translation: AAM51153.1.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1G5AX-ray1.40A13-636[»]
1JG9X-ray1.66A13-636[»]
1JGIX-ray2.00A13-636[»]
1MVYX-ray2.00A13-636[»]
1MW0X-ray2.01A13-636[»]
1MW1X-ray2.10A13-636[»]
1MW2X-ray2.10A13-636[»]
1MW3X-ray2.00A13-636[»]
1S46X-ray2.20A13-636[»]
1ZS2X-ray2.16A13-636[»]
3UEQX-ray1.85A13-636[»]
4FLOX-ray2.20A13-636[»]
4FLQX-ray2.50A13-636[»]
4FLRX-ray2.40A13-636[»]
4FLSX-ray2.30A13-636[»]
ProteinModelPortaliQ9ZEU2.
SMRiQ9ZEU2. Positions 13-636.
ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

STRINGi546267.NEIPOLOT_01155.

Protein family/group databases

CAZyiGH13. Glycoside Hydrolase Family 13.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Phylogenomic databases

eggNOGiENOG4107QW3. Bacteria.
COG0366. LUCA.

Enzyme and pathway databases

BRENDAi2.4.1.4. 3596.
SABIO-RKQ9ZEU2.

Miscellaneous databases

EvolutionaryTraceiQ9ZEU2.

Family and domain databases

Gene3Di2.60.40.1180. 1 hit.
3.20.20.80. 2 hits.
InterProiIPR015902. Glyco_hydro_13.
IPR006047. Glyco_hydro_13_cat_dom.
IPR013780. Glyco_hydro_b.
IPR013781. Glyco_hydro_catalytic_dom.
IPR017853. Glycoside_hydrolase_SF.
[Graphical view]
PANTHERiPTHR10357. PTHR10357. 2 hits.
PfamiPF00128. Alpha-amylase. 1 hit.
[Graphical view]
SMARTiSM00642. Aamy. 1 hit.
[Graphical view]
SUPFAMiSSF51445. SSF51445. 1 hit.
ProtoNetiSearch...

Publicationsi

  1. "Cloning and characterization of the gene for amylosucrase from Neisseria polysaccharea: production of a linear alpha-1,4-glucan."
    Buettcher V., Welsh T., Willmitzer L., Kossmann J.
    J. Bacteriol. 179:3324-3330(1997) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF N-TERMINUS, SUBCELLULAR LOCATION.
    Strain: ATCC 43768 / CCUG 18030 / CIP 100113 / NCTC 11858.
  2. "Sequence analysis of the gene encoding amylosucrase from Neisseria polysaccharea and characterization of the recombinant enzyme."
    Potocki de Montalk G., Remaud-Simeon M., Willemot R.-M., Planchot V., Monsan P.
    J. Bacteriol. 181:375-381(1999) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, SUBUNIT.
    Strain: ATCC 43768 / CCUG 18030 / CIP 100113 / NCTC 11858.
  3. "Nonencapsulated Neisseria meningitidis strain produces amylopectin from sucrose: altering the concept for differentiation between N. meningitidis and N. polysaccharea."
    Zhu P., Tsang R.S.W., Tsai C.-M.
    J. Clin. Microbiol. 41:273-278(2003) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    Strain: 85322.
  4. "Amylosucrase from Neisseria polysaccharea: novel catalytic properties."
    Potocki de Montalk G., Remaud-Simeon M., Willemot R.-M., Sarcabal P., Planchot V., Monsan P.
    FEBS Lett. 471:219-223(2000) [PubMed] [Europe PMC] [Abstract]
    Cited for: CHARACTERIZATION, KINETIC PARAMETERS, CATALYTIC ACTIVITY.
    Strain: ATCC 43768 / CCUG 18030 / CIP 100113 / NCTC 11858.
  5. "Identification of key amino acid residues in Neisseria polysaccharea amylosucrase."
    Sarcabal P., Remaud-Simeon M., Willemot R.-M., Potocki de Montalk G., Svensson B., Monsan P.
    FEBS Lett. 474:33-37(2000) [PubMed] [Europe PMC] [Abstract]
    Cited for: CATALYTIC ACTIVITY, MUTAGENESIS OF HIS-195; ASP-294; GLU-308; GLU-336; GLU-352; HIS-400 AND ASP-401, ACTIVE SITE.
    Strain: ATCC 43768 / CCUG 18030 / CIP 100113 / NCTC 11858.
  6. "Crystal structures of amylosucrase from Neisseria polysaccharea in complex with D-glucose and the active site mutant Glu328Gln in complex with the natural substrate sucrose."
    Mirza O., Skov L.K., Remaud-Simeon M., Potocki de Montalk G., Albenne C., Monsan P., Gajhede M.
    Biochemistry 40:9032-9039(2001) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.66 ANGSTROMS) OF 13-636 IN COMPLEX WITH D-GLUCOSE AND SUCROSE.
    Strain: ATCC 43768 / CCUG 18030 / CIP 100113 / NCTC 11858.
  7. Cited for: X-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS) OF 13-636.
    Strain: ATCC 43768 / CCUG 18030 / CIP 100113 / NCTC 11858.
  8. "Oligosaccharide and sucrose complexes of amylosucrase. Structural implications for the polymerase activity."
    Skov L.K., Mirza O., Sprogoe D., Dar I., Remaud-Simeon M., Albenne C., Monsan P., Gajhede M.
    J. Biol. Chem. 277:47741-47747(2002) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 13-636 OF MUTANT GLN-336 IN COMPLEX WITH SUCROSE AND MALTOHEPTAOSE.
  9. "Crystal structure of the covalent intermediate of amylosucrase from Neisseria polysaccharea."
    Jensen M.H., Mirza O., Albenne C., Remaud-Simeon M., Monsan P., Gajhede M., Skov L.K.
    Biochemistry 43:3104-3110(2004) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 13-636 OF MUTANT GLN-336 IN COMPLEX WITH ALPHA-D-GLUCOPYRANOSYL FLUORIDE, ACTIVE SITE.

Entry informationi

Entry nameiAMYS_NEIPO
AccessioniPrimary (citable) accession number: Q9ZEU2
Secondary accession number(s): Q84HD5
Entry historyi
Integrated into UniProtKB/Swiss-Prot: December 20, 2005
Last sequence update: May 1, 1999
Last modified: July 6, 2016
This is version 80 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Miscellaneous

Like the recombinant protein expressed in E.coli, the amylosucrase may be secreted in N.polysaccharea without cleavage of a signal sequence.

Keywords - Technical termi

3D-structure, Direct protein sequencing

Documents

  1. Glycosyl hydrolases
    Classification of glycosyl hydrolase families and list of entries
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  3. SIMILARITY comments
    Index of protein domains and families

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.