ID   E3GAL_FLAVE             Reviewed;         249 AA.
AC   F7J1C8;
DT   03-APR-2013, integrated into UniProtKB/Swiss-Prot.
DT   21-SEP-2011, sequence version 1.
DT   03-AUG-2022, entry version 32.
DE   RecName: Full=Galactan endo-beta-1,3-galactanase;
DE            EC=3.2.1.181;
DE   AltName: Full=Endo-beta-1,3-galactanase;
DE            Short=FvEN3GAL;
DE   Flags: Precursor;
GN   Name=EN3GAL;
OS   Flammulina velutipes (Agaricus velutipes).
OC   Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes;
OC   Agaricomycetidae; Agaricales; Physalacriaceae; Flammulina.
OX   NCBI_TaxID=38945;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 22-31, FUNCTION, CATALYTIC
RP   ACTIVITY, ACTIVE SITE, GLYCOSYLATION, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP   MUTAGENESIS OF GLU-138 AND GLU-143.
RX   PubMed=21653698; DOI=10.1074/jbc.m111.251736;
RA   Kotake T., Hirata N., Degi Y., Ishiguro M., Kitazawa K., Takata R.,
RA   Ichinose H., Kaneko S., Igarashi K., Samejima M., Tsumuraya Y.;
RT   "Endo-beta-1,3-galactanase from winter mushroom Flammulina velutipes.";
RL   J. Biol. Chem. 286:27848-27854(2011).
CC   -!- FUNCTION: Specifically hydrolyzes beta-1,3-galactan in an endo-fashion.
CC       Requires at least 3 contiguous beta-1,3-residues.
CC       {ECO:0000269|PubMed:21653698}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=The enzyme specifically hydrolyzes beta-1,3-galactan and beta-
CC         1,3-galactooligosaccharides.; EC=3.2.1.181;
CC         Evidence={ECO:0000269|PubMed:21653698};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=0.137 mM for beta-1,3-galactan {ECO:0000269|PubMed:21653698};
CC         KM=1.25 mM for beta-1,3-galactotriose {ECO:0000269|PubMed:21653698};
CC         KM=0.57 mM for beta-1,3-galactotetraose
CC         {ECO:0000269|PubMed:21653698};
CC         Note=kcat is 7.56 sec(-1) with beta-1,3-galactan. kcat is 5.47 sec(-
CC         1) with beta-1,3-galactotriose. kcat is 6.50 sec(-1) with beta-1,3-
CC         galactotetraose.;
CC       pH dependence:
CC         Optimum pH is 4.0-6.0. {ECO:0000269|PubMed:21653698};
CC       Temperature dependence:
CC         Optimum temperature is 50 degrees Celsius.
CC         {ECO:0000269|PubMed:21653698};
CC   -!- PTM: N-glycosylated. {ECO:0000269|PubMed:21653698}.
CC   -!- SIMILARITY: Belongs to the glycosyl hydrolase 16 family. {ECO:0000305}.
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DR   EMBL; AB610981; BAK48741.1; -; mRNA.
DR   AlphaFoldDB; F7J1C8; -.
DR   SMR; F7J1C8; -.
DR   CAZy; GH16; Glycoside Hydrolase Family 16.
DR   CLAE; EBG16A_FLAVE; -.
DR   KEGG; ag:BAK48741; -.
DR   BioCyc; MetaCyc:MON-17420; -.
DR   BRENDA; 3.2.1.181; 1577.
DR   GO; GO:0004553; F:hydrolase activity, hydrolyzing O-glycosyl compounds; IDA:UniProtKB.
DR   GO; GO:0005975; P:carbohydrate metabolic process; IDA:UniProtKB.
DR   InterPro; IPR013320; ConA-like_dom_sf.
DR   InterPro; IPR000757; GH16.
DR   SUPFAM; SSF49899; SSF49899; 1.
DR   PROSITE; PS51762; GH16_2; 1.
PE   1: Evidence at protein level;
KW   Direct protein sequencing; Glycoprotein; Glycosidase; Hydrolase; Signal.
FT   SIGNAL          1..21
FT                   /evidence="ECO:0000269|PubMed:21653698"
FT   CHAIN           22..249
FT                   /note="Galactan endo-beta-1,3-galactanase"
FT                   /id="PRO_0000422019"
FT   DOMAIN          34..247
FT                   /note="GH16"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01098"
FT   ACT_SITE        138
FT                   /note="Nucleophile"
FT                   /evidence="ECO:0000305|PubMed:21653698"
FT   ACT_SITE        143
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000305|PubMed:21653698"
FT   CARBOHYD        48
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   CARBOHYD        156
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   MUTAGEN         138
FT                   /note="E->D: Loss of galactan endo-beta-1,3-galactanase
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:21653698"
FT   MUTAGEN         143
FT                   /note="E->D: Loss of galactan endo-beta-1,3-galactanase
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:21653698"
SQ   SEQUENCE   249 AA;  26690 MW;  6FB4218B8D52F1B6 CRC64;
     MKLFVVLACL AAPGTFPFVD AATVIPANSF SSFSTYWNNF YPWGTDHNGS GRMASANIIV
     ASNTLSLIAT PTSNPSPPTS TSNPKPAIHY ASGAIHAKEH ITVTAANAYT VSGEFSAPTA
     VGTWPAFWLT AVSGWPPEVD IGEWKGTADN WFNTFNTSSV VKSTLVDWPT DLSFHSVKAV
     LTAQSNNKDV KIDFYMDNKF IVTQYGSGFV GKAMYLIINL QMEGSSGSPG PSGRTVYKAR
     NVQVTRTGN