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Protein

Mannan endo-1,4-beta-mannosidase

Gene

manA

Organism
Cellvibrio japonicus (strain Ueda107) (Pseudomonas fluorescens subsp. cellulosa)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the endo hydrolysis of beta-1,4-linked mannan and galactomannan, but displays little activity towards other polysaccharides located in the plant cell wall (PubMed:11382747). Preferentially hydrolyzes the larger oligosaccharides and has greater activity against non-substituted polysaccharides (PubMed:7848261) (PubMed:8973192). It displays tight specificity for mannose at both the -2 and the -1 subsites (PubMed:19441796). Appears to act in synergy with alpha-galactosidase (AgaA) to elicit hydrolysis of galactomannan (PubMed:11064195).5 Publications

Catalytic activityi

Random hydrolysis of (1->4)-beta-D-mannosidic linkages in mannans, galactomannans and glucomannans.4 Publications

Kineticsi

Kcat is 14.7 sec(-1) for mannanase activity with 2,4-dinitrophenyl-beta-mannobioside (2,4-DNPM) as substrate (PubMed:8973192). Kcat is 20 sec(-1) for mannanase activity with 2,4-dinitrophenyl-beta-mannobioside (2,4-DNPM) as substrate (PubMed:11382747). Kcat is 1759 sec(-1) for mannanase activity with carob galactomannan as substrate (PubMed:8973192). Kcat is 2904 sec(-1) for mannanase activity with carob galactomannan as substrate (PubMed:11382747). Kcat is 3861 sec(-1) for mannanase activity with mannotetraose as substrate (PubMed:8973192). Kcat is 280000 min(-1) for mannanase activity with galactomannan as substrate (PubMed:19441796). Kcat is 400000 min(-1) for mannanase activity with glucomannan as substrate (PubMed:19441796).3 Publications

  1. KM=0.9 mg/ml for carob galactomannan1 Publication
  2. KM=2.8 mg/ml for glucomannan1 Publication
  3. KM=3.2 mg/ml for galactomannan1 Publication
  4. KM=9.3 µM for 2,4-dinitrophenyl-beta-mannobioside (2,4-DNPM)1 Publication
  5. KM=12.3 µM for 2,4-dinitrophenyl-beta-mannobioside (2,4-DNPM)1 Publication

    pH dependencei

    Optimum pH is about 7.0.1 Publication

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei121 – 1211Substrate2 Publications
    Binding sitei143 – 1431Substrate3 Publications
    Binding sitei162 – 1621Substrate1 Publication1 Publication
    Sitei211 – 2111Plays an important role in maintaining the position of the catalytic nucleophile1 Publication1 Publication
    Active sitei212 – 2121Proton donor4 Publications
    Binding sitei217 – 2171Substrate1 Publication
    Binding sitei285 – 2851Substrate2 Publications
    Active sitei320 – 3201Nucleophile2 Publications
    Binding sitei377 – 3771Substrate3 Publications

    GO - Molecular functioni

    • mannan endo-1,4-beta-mannosidase activity Source: UniProtKB

    GO - Biological processi

    • galactomannan metabolic process Source: UniProtKB
    • glucomannan metabolic process Source: UniProtKB
    • polysaccharide catabolic process Source: UniProtKB-KW
    Complete GO annotation...

    Keywords - Molecular functioni

    Glycosidase, Hydrolase

    Keywords - Biological processi

    Carbohydrate metabolism, Polysaccharide degradation

    Enzyme and pathway databases

    BioCyciCJAP498211:GHIT-2761-MONOMER.

    Protein family/group databases

    CAZyiGH26. Glycoside Hydrolase Family 26.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Mannan endo-1,4-beta-mannosidase1 Publication (EC:3.2.1.784 Publications)
    Alternative name(s):
    Mannanase 26A1 Publication
    Short name:
    Man26A1 Publication
    Mannanase A1 Publication
    Short name:
    ManA1 Publication
    Gene namesi
    Name:manA1 Publication
    Synonyms:man26A
    Ordered Locus Names:CJA_2770
    OrganismiCellvibrio japonicus (strain Ueda107) (Pseudomonas fluorescens subsp. cellulosa)
    Taxonomic identifieri498211 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaCellvibrionalesCellvibrionaceaeCellvibrio
    Proteomesi
    • UP000001036 Componenti: Chromosome

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi121 – 1211E → A: Strong decrease of mannanase activity against both oligosaccharides and polysaccharides. 1 Publication
    Mutagenesisi143 – 1431H → A: Very low mannanase activity against both mannans and mannooligosaccharides. 1 Publication
    Mutagenesisi156 – 1561W → A: Similar to the wild-type. 1 Publication
    Mutagenesisi162 – 1621W → A: The mannanase activity is 95, 70, and 30-fold less active than the wild-type against mannotriose, mannotetraose and mannohexaose, respectively. 1 Publication
    Mutagenesisi211 – 2111H → A: Causes a 100- and 700-fold decrease in mannanase activity against carob galactomannans and mannotetraose, respectively, but only a 6-fold reduction in mannanase activity against 2,4-DNPM. 1 Publication
    Mutagenesisi212 – 2121E → A: Causes a dramatic decrease in the catalytic efficiency against carob galactomannan, azo-carob galactomannan, mannotetraose and 2,4-DNPM. 2 Publications
    Mutagenesisi212 – 2121E → D: 10-fold decrease of the catalytic efficiency against 2,4-DNPM. 1 Publication
    Mutagenesisi217 – 2171W → A: Lack of mannanase activity against both mannotriose and mannotetraose. 1 Publication
    Mutagenesisi278 – 2781D → A: Retains significant activity against mannotetraose and azo-carob galactomannan. 1 Publication
    Mutagenesisi278 – 2781D → E: Retains significant activity against mannotetraose and azo-carob galactomannan. 1 Publication
    Mutagenesisi283 – 2831D → A: Retains significant activity against mannotetraose and azo-carob galactomannan. Lack of activity against all substrates; when associated with A-320. 2 Publications
    Mutagenesisi283 – 2831D → E: Retains significant activity against mannotetraose and azo-carob galactomannan. 1 Publication
    Mutagenesisi285 – 2851Y → A: Reduction of the catalytic efficiency with carob galactomannan and 2,4-DNPM. 1 Publication
    Mutagenesisi320 – 3201E → A: Does not alter the affinity against carob galactomannan, azo-carob galactomannan, mannotetraose and 2,4-DNPM. Lack of activity against all substrates; when associated with A-283. 2 Publications
    Mutagenesisi320 – 3201E → D: Causes a dramatic decrease in the catalytic efficiency against mannotetraose. 1 Publication
    Mutagenesisi320 – 3201E → G: Lack of activity against all substrates. 2 Publications
    Mutagenesisi360 – 3601W → A: Lack of activity against all substrates. 1 Publication
    Mutagenesisi361 – 3611R → A: Strong decrease of mannanase activity against oligosaccharides, while the reduction in the rate of polysaccharide hydrolysis is more modest. Strong decrease of mannanase activity and not able to distinguish between mannose and glucose units; when associated with A-377. 1 Publication
    Mutagenesisi377 – 3771H → A: 100-fold decrease in the activity of the mannanase against mannotetraose and 4-nitrophenyl-beta-D-Man2. It hydrolyzes polysaccharides only 10-20-fold less efficiently than the wild-type. Strong decrease of mannanase activity and not able to distinguish between mannose and glucose units; when associated with A-361. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Signal peptidei1 – 2727PROSITE-ProRule annotation1 PublicationAdd
    BLAST
    Chaini28 – 423396Mannan endo-1,4-beta-mannosidasePRO_0000012173Add
    BLAST

    Interactioni

    Subunit structurei

    Homodimer.1 Publication

    Protein-protein interaction databases

    STRINGi498211.CJA_2770.

    Structurei

    Secondary structure

    1
    423
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi45 – 473Combined sources
    Helixi56 – 6813Combined sources
    Turni69 – 713Combined sources
    Beta strandi73 – 786Combined sources
    Turni79 – 824Combined sources
    Beta strandi91 – 933Combined sources
    Helixi96 – 1016Combined sources
    Beta strandi106 – 1116Combined sources
    Helixi112 – 1143Combined sources
    Helixi125 – 1339Combined sources
    Beta strandi137 – 1415Combined sources
    Turni147 – 1493Combined sources
    Helixi150 – 1523Combined sources
    Turni156 – 1594Combined sources
    Turni169 – 1713Combined sources
    Helixi178 – 19316Combined sources
    Beta strandi205 – 2084Combined sources
    Beta strandi215 – 2184Combined sources
    Turni222 – 2243Combined sources
    Helixi227 – 24216Combined sources
    Beta strandi249 – 2546Combined sources
    Helixi264 – 2685Combined sources
    Turni274 – 2763Combined sources
    Beta strandi278 – 2803Combined sources
    Beta strandi283 – 2864Combined sources
    Beta strandi288 – 2903Combined sources
    Helixi292 – 31221Combined sources
    Beta strandi320 – 3234Combined sources
    Helixi325 – 3295Combined sources
    Helixi337 – 34711Combined sources
    Helixi351 – 3533Combined sources
    Beta strandi355 – 3595Combined sources
    Helixi385 – 3895Combined sources
    Helixi392 – 40110Combined sources
    Helixi408 – 4103Combined sources
    Beta strandi419 – 4213Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1GVYX-ray1.70A39-363[»]
    1GW1X-ray1.65A43-421[»]
    1J9YX-ray1.85A39-423[»]
    1ODZX-ray1.40A/B39-423[»]
    1R7OX-ray1.85A29-423[»]
    2WHMX-ray1.50A39-423[»]
    ProteinModelPortaliP49424.
    SMRiP49424. Positions 42-422.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP49424.

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini56 – 409354GH26PROSITE-ProRule annotationAdd
    BLAST

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni360 – 3612Substrate binding3 Publications

    Sequence similaritiesi

    Belongs to the glycosyl hydrolase 26 family.PROSITE-ProRule annotationCurated
    Contains 1 GH26 (glycosyl hydrolase family 26) domain.PROSITE-ProRule annotation

    Keywords - Domaini

    Signal

    Phylogenomic databases

    eggNOGiENOG4105D3P. Bacteria.
    COG4124. LUCA.
    HOGENOMiHOG000274384.
    KOiK01218.
    OMAiFLYAYSP.
    OrthoDBiEOG6V4G9N.

    Family and domain databases

    Gene3Di3.20.20.80. 1 hit.
    InterProiIPR022790. GH26_dom.
    IPR000805. Glyco_hydro_26.
    IPR013781. Glyco_hydro_catalytic_dom.
    IPR017853. Glycoside_hydrolase_SF.
    [Graphical view]
    PANTHERiPTHR40079. PTHR40079. 1 hit.
    PfamiPF02156. Glyco_hydro_26. 1 hit.
    [Graphical view]
    PRINTSiPR00739. GLHYDRLASE26.
    SUPFAMiSSF51445. SSF51445. 1 hit.
    PROSITEiPS51764. GH26. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    P49424-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MKTITTARLP WAAQSFALGI CLIALLGCNH AANKSSASRA DVKPVTVKLV
    60 70 80 90 100
    DSQATMETRS LFAFMQEQRR HSIMFGHQHE TTQGLTITRT DGTQSDTFNA
    110 120 130 140 150
    VGDFAAVYGW DTLSIVAPKA EGDIVAQVKK AYARGGIITV SSHFDNPKTD
    160 170 180 190 200
    TQKGVWPVGT SWDQTPAVVD SLPGGAYNPV LNGYLDQVAE WANNLKDEQG
    210 220 230 240 250
    RLIPVIFRLY HENTGSWFWW GDKQSTPEQY KQLFRYSVEY LRDVKGVRNF
    260 270 280 290 300
    LYAYSPNNFW DVTEANYLER YPGDEWVDVL GFDTYGPVAD NADWFRNVVA
    310 320 330 340 350
    NAALVARMAE ARGKIPVISE IGIRAPDIEA GLYDNQWYRK LISGLKADPD
    360 370 380 390 400
    AREIAFLLVW RNAPQGVPGP NGTQVPHYWV PANRPENINN GTLEDFQAFY
    410 420
    ADEFTAFNRD IEQVYQRPTL IVK
    Length:423
    Mass (Da):47,487
    Last modified:July 5, 2005 - v2
    Checksum:iEFDAB597F10BA3DD
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    X82179 Genomic DNA. Translation: CAA57670.2.
    CP000934 Genomic DNA. Translation: ACE82849.1.
    PIRiS53374.
    RefSeqiWP_012488364.1. NC_010995.1.

    Genome annotation databases

    EnsemblBacteriaiACE82849; ACE82849; CJA_2770.
    KEGGicja:CJA_2770.
    PATRICi21328934. VBICelJap122165_2735.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    X82179 Genomic DNA. Translation: CAA57670.2.
    CP000934 Genomic DNA. Translation: ACE82849.1.
    PIRiS53374.
    RefSeqiWP_012488364.1. NC_010995.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1GVYX-ray1.70A39-363[»]
    1GW1X-ray1.65A43-421[»]
    1J9YX-ray1.85A39-423[»]
    1ODZX-ray1.40A/B39-423[»]
    1R7OX-ray1.85A29-423[»]
    2WHMX-ray1.50A39-423[»]
    ProteinModelPortaliP49424.
    SMRiP49424. Positions 42-422.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi498211.CJA_2770.

    Protein family/group databases

    CAZyiGH26. Glycoside Hydrolase Family 26.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiACE82849; ACE82849; CJA_2770.
    KEGGicja:CJA_2770.
    PATRICi21328934. VBICelJap122165_2735.

    Phylogenomic databases

    eggNOGiENOG4105D3P. Bacteria.
    COG4124. LUCA.
    HOGENOMiHOG000274384.
    KOiK01218.
    OMAiFLYAYSP.
    OrthoDBiEOG6V4G9N.

    Enzyme and pathway databases

    BioCyciCJAP498211:GHIT-2761-MONOMER.

    Miscellaneous databases

    EvolutionaryTraceiP49424.

    Family and domain databases

    Gene3Di3.20.20.80. 1 hit.
    InterProiIPR022790. GH26_dom.
    IPR000805. Glyco_hydro_26.
    IPR013781. Glyco_hydro_catalytic_dom.
    IPR017853. Glycoside_hydrolase_SF.
    [Graphical view]
    PANTHERiPTHR40079. PTHR40079. 1 hit.
    PfamiPF02156. Glyco_hydro_26. 1 hit.
    [Graphical view]
    PRINTSiPR00739. GLHYDRLASE26.
    SUPFAMiSSF51445. SSF51445. 1 hit.
    PROSITEiPS51764. GH26. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "A non-modular endo-beta-1,4-mannanase from Pseudomonas fluorescens subspecies cellulosa."
      Braithwaite K.L., Black G.W., Hazlewood G.P., Ali B.R.S., Gilbert H.J.
      Biochem. J. 305:1005-1010(1995) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 39-48, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY.
    2. Gilbert H.J.
      Submitted (JUL-2001) to the EMBL/GenBank/DDBJ databases
      Cited for: SEQUENCE REVISION TO 364-423.
    3. "Insights into plant cell wall degradation from the genome sequence of the soil bacterium Cellvibrio japonicus."
      DeBoy R.T., Mongodin E.F., Fouts D.E., Tailford L.E., Khouri H., Emerson J.B., Mohamoud Y., Watkins K., Henrissat B., Gilbert H.J., Nelson K.E.
      J. Bacteriol. 190:5455-5463(2008) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: Ueda107.
    4. "Mannanase A from Pseudomonas fluorescens ssp. cellulosa is a retaining glycosyl hydrolase in which E212 and E320 are the putative catalytic residues."
      Bolam D.N., Hughes N., Virden R., Lakey J.H., Hazlewood G.P., Henrissat B., Braithwaite K.L., Gilbert H.J.
      Biochemistry 35:16195-16204(1996) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF GLU-212; ASP-278; ASP-283 AND GLU-320, BIOPHYSICOCHEMICAL PROPERTIES, REACTION MECHANISM.
    5. "Alpha-galactosidase A from Pseudomonas fluorescens subsp. cellulosa: cloning, high level expression and its role in galactomannan hydrolysis."
      Halstead J.R., Fransen M.P., Eberhart R.Y., Park A.J., Gilbert H.J., Hazlewood G.P.
      FEMS Microbiol. Lett. 192:197-203(2000) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION.
    6. "Crystal structure of mannanase 26A from Pseudomonas cellulosa and analysis of residues involved in substrate binding."
      Hogg D., Woo E.J., Bolam D.N., McKie V.A., Gilbert H.J., Pickersgill R.W.
      J. Biol. Chem. 276:31186-31192(2001) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF HIS-143; TRP-156; TRP-162; HIS-211; TRP-217; ASP-283; TYR-285; GLU-320 AND TRP-360.
    7. "Substrate distortion by a beta-mannanase: snapshots of the Michaelis and covalent-intermediate complexes suggest a B(2,5) conformation for the transition state."
      Ducros V.M., Zechel D.L., Murshudov G.N., Gilbert H.J., Szabo L., Stoll D., Withers S.G., Davies G.J.
      Angew. Chem. Int. Ed. Engl. 41:2824-2827(2002) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF MUTANT ALA-212 OF 39-421 IN COMPLEX WITH SUBSTRATE ANALOGS, MUTAGENESIS OF GLU-212.
    8. "Expansion of the glycosynthase repertoire to produce defined manno-oligosaccharides."
      Jahn M., Stoll D., Warren R.A., Szabo L., Singh P., Gilbert H.J., Ducros V.M., Davies G.J., Withers S.G.
      Chem. Commun. (Camb.) 2003:1327-1329(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) MUTANT GLY-320 OF 39-423 IN COMPLEX WITH SUBSTRATE ANALOGS, MUTAGENESIS OF GLU-320, SUBUNIT.
    9. "Structural investigation of Mannanase 26A from Pseudomonas cellulosa reveals an induced fit mechanism and a non-substrate ligand binding site."
      Oakley A.J., Wilce M.C.J.
      Submitted (OCT-2003) to the PDB data bank
      Cited for: X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) OF OF 29-423.
    10. Cited for: X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF DOUBLE MUTANT ALA-121/GLY-320 OF 39-423 IN COMPLEX WITH SUBSTRATE ANALOGS, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF GLU-121; GLU-320; ARG-361 AND HIS-377, SUBSTRATE SPECIFICITY.

    Entry informationi

    Entry nameiMANA_CELJU
    AccessioniPrimary (citable) accession number: P49424
    Secondary accession number(s): B3PBK3
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: February 1, 1996
    Last sequence update: July 5, 2005
    Last modified: March 16, 2016
    This is version 92 of the entry and version 2 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Complete proteome, Direct protein sequencing, Reference proteome

    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.