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Information on EC 3.1.1.86 - rhamnogalacturonan acetylesterase and Organism(s) Aspergillus aculeatus and UniProt Accession Q00017

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EC Tree
     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.1 Carboxylic-ester hydrolases
                3.1.1.86 rhamnogalacturonan acetylesterase
IUBMB Comments
The degradation of rhamnogalacturonan by rhamnogalacturonases depends on the removal of the acetyl esters from the substrate .
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This record set is specific for:
Aspergillus aculeatus
UNIPROT: Q00017
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Word Map
The taxonomic range for the selected organisms is: Aspergillus aculeatus
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
Hydrolytic cleavage of 2-O-acetyl- or 3-O-acetyl groups of alpha-D-galacturonic acid in rhamnogalacturonan I.
Synonyms
rhamnogalacturonan acetylesterase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
rhamnogalacturonan 2/3-O-acetyl-alpha-D-galacturonate O-acetylhydrolase
The degradation of rhamnogalacturonan by rhamnogalacturonases depends on the removal of the acetyl esters from the substrate [1].
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetylated rhamnogalacturonan I + H2O
acetate + deacetylated rhamnogalacturonan I
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetylated rhamnogalacturonan I + H2O
acetate + deacetylated rhamnogalacturonan I
show the reaction diagram
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 6
upon prolonged incubation the recombinant enzyme hydrolyses acetyl groups from modified hairy regions from apples to the same extent at pH 3.5 as at pH 6.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
RHA1_ASPAC
250
0
26351
Swiss-Prot
Secretory Pathway (Reliability: 1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26350
x * 26350, calculated from sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 26350, calculated from sequence
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure refined to a resolution of 1.12 A using synchrotron data collected at 263 K
crystallized in two crystal forms, an orthorhombic and a trigonal crystal form. In the orthorhombic crystal form, the covalently bound carbohydrate at one of the two N-glycosylation sites is involved in crystal contacts. The orthorhombic crystal form is obtained at pH 5.0 and the trigonal crystal form at pH 4.5
crystals used for structure determination are grown in 1.4 M Li2SO4, 0.1 M Na acetate pH 5.0 with a protein concentration of 40 OD280. The crystals belong to the space group P212121 with a = 52.14 A, b = 56.87 A and c = 71.89 A and one molecule in the asymmetric unit.The structure of RGAE is determined at 1.55 A resolution. RGAE folds into an alpha/beta/alpha structure. The active site of RGAE is an open cleft containing a serine-histidine-aspartic acid catalytic triad. The position of the three residues relative to the central parallel beta sheet and the lack of the nucleophilic elbow motif found in structures possessing the alpha/beta hydrolase fold shows that RGAE does not belong to the alpha/beta hydrolase family
hanging-drop vapour-diffusion method
vapour-diffusion method, the crystal structure of rhamnogalacturonan acetylesterase D192N is determined to 1.33 A resolution and refined to an R value of 11.6% for all data. The structure is virtually identical to the high-resolution (1.12 A) structure of the wild-type enzyme except for the interactions involving the mutation and a disordered loop
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
overexpressed in Aspergillus oryzae
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the expression of rhamnogalacturonan degrading enzymes by Aspergillus aculeatus is regulated at the level of transcription and is subjected to carbon catabolite repression by glucose
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Molgaard, A.; Petersen, J.F.; Kauppinen, S.; Dalboge, H.; Johnsen, A.H.; Navarro Poulsen, J.C.
Larsen, S.: Crystallization and preliminary X-ray diffraction studies of the heterogeneously glycosylated enzyme rhamnogalacturonan acetylesterase from Aspergillus aculeatus
Acta Crystallogr. Sect. D
54
1026-1029
1998
Aspergillus aculeatus (Q00017), Aspergillus aculeatus
Manually annotated by BRENDA team
Molgaard, A.; Larsen, S.
A branched N-linked glycan at atomic resolution in the 1.12 A structure of rhamnogalacturonan acetylesterase
Acta Crystallogr. Sect. D
58
111-119
2001
Aspergillus aculeatus (Q00017), Aspergillus aculeatus
Manually annotated by BRENDA team
Molgaard, A.; Larsen, S.
Crystal packing in two pH-dependent crystal forms of rhamnogalacturonan acetylesterase
Acta Crystallogr. Sect. D
60
472-478
2004
Aspergillus aculeatus (Q00017), Aspergillus aculeatus
Manually annotated by BRENDA team
Langkilde, A.; Kristensen, S.M.; Lo Leggio, L.; Molgaard, A.; Jensen, J.H.; Houk, A.R.; Navarro Poulsen, J.C.; Kauppinen, S.; Larsen, S.
Short strong hydrogen bonds in proteins: a case study of rhamnogalacturonan acetylesterase
Acta Crystallogr. Sect. D
64
851-863
2008
Aspergillus aculeatus (Q00017)
Manually annotated by BRENDA team
Kauppinen, S.; Christgau, S.; Kofod, L.V.; Halkier, T.; Drreich, K.; Dalboge, H.
Molecular cloning and characterization of a rhamnogalacturonan acetylesterase from Aspergillus aculeatus. Synergism between rhamnogalacturonan degrading enzymes
J. Biol. Chem.
270
27172-17178
1995
Aspergillus aculeatus (Q00017), Aspergillus aculeatus
Manually annotated by BRENDA team
Molgaard, A.; Kauppinen, S.; Larsen, S.
Rhamnogalacturonan acetylesterase elucidates the structure and function of a new family of hydrolases
Structure
8
373-383
2000
Aspergillus aculeatus (Q00017), Aspergillus aculeatus
Manually annotated by BRENDA team