EABN2_BACSU
ID EABN2_BACSU Reviewed; 469 AA.
AC P42293; B3FRL6;
DT 01-NOV-1995, integrated into UniProtKB/Swiss-Prot.
DT 28-JUL-2009, sequence version 2.
DT 03-AUG-2022, entry version 129.
DE RecName: Full=Extracellular endo-alpha-(1->5)-L-arabinanase 2;
DE Short=ABN;
DE EC=3.2.1.99;
DE AltName: Full=Endo-1,5-alpha-L-arabinanase;
DE Flags: Precursor;
GN Name=abn2; Synonyms=J3A, yxiA; OrderedLocusNames=BSU39330;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168 / BGSC1A1;
RX PubMed=7704263; DOI=10.1099/13500872-141-2-337;
RA Yoshida K., Sano H., Seki S., Oda M., Fujimura M., Fujita Y.;
RT "Cloning and sequencing of a 29 kb region of the Bacillus subtilis genome
RT containing the hut and wapA loci.";
RL Microbiology 141:337-343(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 27-31, FUNCTION,
RP DISRUPTION PHENOTYPE, BIOPHYSICOCHEMICAL PROPERTIES, INDUCTION, SUBCELLULAR
RP LOCATION, SUBSTRATE SPECIFICITY, AND NOMENCLATURE.
RC STRAIN=168;
RX PubMed=18408032; DOI=10.1128/jb.00162-08;
RA Inacio J.M., de Sa-Nogueira I.;
RT "Characterization of abn2 (yxiA), encoding a Bacillus subtilis GH43
RT arabinanase, Abn2, and its role in arabino-polysaccharide degradation.";
RL J. Bacteriol. 190:4272-4280(2008).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [4]
RP SEQUENCE REVISION TO 175.
RX PubMed=19383706; DOI=10.1099/mic.0.027839-0;
RA Barbe V., Cruveiller S., Kunst F., Lenoble P., Meurice G., Sekowska A.,
RA Vallenet D., Wang T., Moszer I., Medigue C., Danchin A.;
RT "From a consortium sequence to a unified sequence: the Bacillus subtilis
RT 168 reference genome a decade later.";
RL Microbiology 155:1758-1775(2009).
RN [5]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-171.
RX PubMed=2454913; DOI=10.1128/jb.170.7.3199-3205.1988;
RA Oda M., Sugishita A., Furukawa K.;
RT "Cloning and nucleotide sequences of histidase and regulatory genes in the
RT Bacillus subtilis hut operon and positive regulation of the operon.";
RL J. Bacteriol. 170:3199-3205(1988).
RN [6]
RP SUBCELLULAR LOCATION.
RX PubMed=18607095; DOI=10.1107/s1744309108016321;
RA de Sanctis D., Bento I., Inacio J.M., Custodio S., de Sa-Nogueira I.,
RA Carrondo M.A.;
RT "Overproduction, crystallization and preliminary X-ray characterization of
RT Abn2, an endo-1,5-alpha-arabinanase from Bacillus subtilis.";
RL Acta Crystallogr. F 64:636-638(2008).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF MUTANTS ALA-171 AND HIS-318 IN
RP COMPLEX WITH ALPHA-L-ARABINOFURANOSE AND CALCIUM IONS, FUNCTION, CATALYTIC
RP ACTIVITY, ACTIVE SITE, MUTAGENESIS OF ASP-38; ASP-171; GLU-224 AND HIS-318,
RP SUBCELLULAR LOCATION, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND SUBUNIT.
RX PubMed=20883454; DOI=10.1111/j.1742-4658.2010.07870.x;
RA de Sanctis D., Inacio J.M., Lindley P.F., de Sa-Nogueira I., Bento I.;
RT "New evidence for the role of calcium in the glycosidase reaction of GH43
RT arabinanases.";
RL FEBS J. 277:4562-4574(2010).
CC -!- FUNCTION: Involved in the degradation of arabinan and is a key enzyme
CC in the complete degradation of the plant cell wall. Catalyzes the
CC internal cleavage of alpha-(1->5)-L-arabinofuranosyl residues of the
CC alpha-1,5-L-arabinan to produce arabino-oligosaccharides and L-
CC arabinose. It is also active toward linear branched sugar beet
CC arabinan, and pectin from apple. {ECO:0000269|PubMed:18408032,
CC ECO:0000269|PubMed:20883454}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endohydrolysis of (1->5)-alpha-arabinofuranosidic linkages in
CC (1->5)-arabinans.; EC=3.2.1.99;
CC Evidence={ECO:0000269|PubMed:20883454};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108; Evidence={ECO:0000305};
CC Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000305};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.11 mM for linear-alpha-1,5-L-arabinan
CC {ECO:0000269|PubMed:18408032, ECO:0000269|PubMed:20883454};
CC KM=2 mM for linear-alpha-1,5-L-arabinan (at pH 7.0 and 50 degrees
CC Celsius) {ECO:0000269|PubMed:18408032, ECO:0000269|PubMed:20883454};
CC Vmax=0.25 mmol/min/mg enzyme for linear-alpha-1,5-L-arabinan (at pH
CC 7.0 and 50 degrees Celsius) {ECO:0000269|PubMed:18408032,
CC ECO:0000269|PubMed:20883454};
CC pH dependence:
CC Optimum pH is 7. {ECO:0000269|PubMed:18408032,
CC ECO:0000269|PubMed:20883454};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius. It remains fully active
CC after 30 minutes of preincubation at 50 degrees Celsius, however,
CC after preincubation at 60 degrees Celsius, the residual activity is
CC only 15%. {ECO:0000269|PubMed:18408032, ECO:0000269|PubMed:20883454};
CC -!- PATHWAY: Glycan metabolism; L-arabinan degradation.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:20883454}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:18408032,
CC ECO:0000269|PubMed:18607095, ECO:0000269|PubMed:20883454}.
CC -!- INDUCTION: Repressed by glucose, and induced by pectin and arabinan.
CC {ECO:0000269|PubMed:18408032}.
CC -!- DISRUPTION PHENOTYPE: No visible phenotype. Double mutant abn2/abnA
CC induce an almost complete loss of this activity.
CC {ECO:0000269|PubMed:18408032}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 43 family. {ECO:0000305}.
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DR EMBL; D31856; BAA06646.1; -; Genomic_DNA.
DR EMBL; EU373814; ACB06752.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB15969.2; -; Genomic_DNA.
DR EMBL; M20659; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR PIR; E70076; E70076.
DR RefSeq; NP_391812.2; NC_000964.3.
DR PDB; 2X8F; X-ray; 1.90 A; A/B=1-469.
DR PDB; 2X8S; X-ray; 1.50 A; A/B=1-469.
DR PDB; 2X8T; X-ray; 1.79 A; A/B=1-469.
DR PDB; 4COT; X-ray; 1.90 A; A=1-469.
DR PDBsum; 2X8F; -.
DR PDBsum; 2X8S; -.
DR PDBsum; 2X8T; -.
DR PDBsum; 4COT; -.
DR AlphaFoldDB; P42293; -.
DR SMR; P42293; -.
DR STRING; 224308.BSU39330; -.
DR CAZy; GH43; Glycoside Hydrolase Family 43.
DR PaxDb; P42293; -.
DR PRIDE; P42293; -.
DR EnsemblBacteria; CAB15969; CAB15969; BSU_39330.
DR GeneID; 937533; -.
DR KEGG; bsu:BSU39330; -.
DR PATRIC; fig|224308.179.peg.4257; -.
DR eggNOG; COG3507; Bacteria.
DR InParanoid; P42293; -.
DR OMA; MNPKTGF; -.
DR PhylomeDB; P42293; -.
DR BioCyc; BSUB:BSU39330-MON; -.
DR BRENDA; 3.2.1.99; 658.
DR UniPathway; UPA00667; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0046558; F:arabinan endo-1,5-alpha-L-arabinosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0031222; P:arabinan catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 2.115.10.20; -; 1.
DR InterPro; IPR032291; Abn2_C.
DR InterPro; IPR006710; Glyco_hydro_43.
DR InterPro; IPR023296; Glyco_hydro_beta-prop_sf.
DR Pfam; PF16369; GH43_C; 1.
DR Pfam; PF04616; Glyco_hydro_43; 1.
DR SUPFAM; SSF75005; SSF75005; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Direct protein sequencing;
KW Glycosidase; Hydrolase; Metal-binding; Reference proteome; Secreted;
KW Signal.
FT SIGNAL 1..26
FT /evidence="ECO:0000255"
FT CHAIN 27..469
FT /note="Extracellular endo-alpha-(1->5)-L-arabinanase 2"
FT /id="PRO_0000012205"
FT ACT_SITE 38
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:20883454"
FT ACT_SITE 224
FT /note="Proton donor"
FT /evidence="ECO:0000269|PubMed:20883454"
FT BINDING 38
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 122
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 168..171
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 188..190
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:20883454"
FT BINDING 220..224
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:20883454"
FT BINDING 318
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000255"
FT SITE 171
FT /note="Important for catalytic activity, responsible for
FT pKa modulation of the active site Glu and correct
FT orientation of both the proton donor and substrate"
FT /evidence="ECO:0000305|PubMed:20883454"
FT SITE 318
FT /note="Important for substrate recognition"
FT MUTAGEN 38
FT /note="D->A: Loss of arabinanase activity."
FT /evidence="ECO:0000269|PubMed:20883454"
FT MUTAGEN 171
FT /note="D->A: Loss of arabinanase activity."
FT /evidence="ECO:0000269|PubMed:20883454"
FT MUTAGEN 224
FT /note="E->A: Loss of arabinanase activity."
FT /evidence="ECO:0000269|PubMed:20883454"
FT MUTAGEN 318
FT /note="H->A: Drastic decrease in arabinanase activity. Does
FT not unduly disrupt the calcium cluster. Mutation does not
FT disrupt the calcium cluster, but it relaxes the geometry of
FT the cluster, resulting in a more planar arrangement of the
FT calcium ion with five of the water molecules."
FT /evidence="ECO:0000269|PubMed:20883454"
FT MUTAGEN 318
FT /note="H->Q: Loss of arabinanase activity."
FT /evidence="ECO:0000269|PubMed:20883454"
FT CONFLICT 175
FT /note="F -> S (in Ref. 1; BAA06646)"
FT /evidence="ECO:0000305"
FT STRAND 40..44
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 47..51
FT /evidence="ECO:0007829|PDB:2X8S"
FT HELIX 53..55
FT /evidence="ECO:0007829|PDB:4COT"
FT STRAND 57..65
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 67..70
FT /evidence="ECO:0007829|PDB:2X8S"
FT HELIX 83..86
FT /evidence="ECO:0007829|PDB:2X8S"
FT HELIX 88..94
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 103..106
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 112..119
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 121..123
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 126..135
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 141..149
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 152..154
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 158..160
FT /evidence="ECO:0007829|PDB:2X8S"
FT TURN 163..165
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 173..176
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 182..186
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 193..198
FT /evidence="ECO:0007829|PDB:2X8S"
FT TURN 200..202
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 203..205
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 213..216
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 222..231
FT /evidence="ECO:0007829|PDB:2X8S"
FT TURN 232..235
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 236..247
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 253..261
FT /evidence="ECO:0007829|PDB:2X8S"
FT HELIX 274..276
FT /evidence="ECO:0007829|PDB:2X8S"
FT HELIX 287..290
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 293..298
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 300..302
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 306..308
FT /evidence="ECO:0007829|PDB:2X8T"
FT STRAND 312..323
FT /evidence="ECO:0007829|PDB:2X8S"
FT TURN 325..327
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 330..337
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 346..354
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 360..362
FT /evidence="ECO:0007829|PDB:2X8S"
FT HELIX 377..380
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 382..388
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 401..405
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 409..415
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 417..422
FT /evidence="ECO:0007829|PDB:2X8S"
FT TURN 423..425
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 426..431
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 434..445
FT /evidence="ECO:0007829|PDB:2X8S"
FT TURN 446..449
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 450..458
FT /evidence="ECO:0007829|PDB:2X8S"
FT STRAND 464..469
FT /evidence="ECO:0007829|PDB:2X8S"
SQ SEQUENCE 469 AA; 52607 MW; 1AB8F3D8EF2B3E2E CRC64;
MFNRLFRVCF LAALIMAFTL PNSVYAQKPI FKEVSVHDPS IIETNGTFYV FGSHLASAKS
NDLMQWQQLT TSVSNDNPLI PNVYEELKET FEWAQSDTLW AADVTQLADG KYYMYYNACR
GDSPRSAMGV AVADNIEGPY KNKGIFLKSG MEGTSSDGTP YDATKHPNVV DPHTFFDKDG
KLWMVYGSYS GGIFILEMNP KTGFPLPGQG YGKKLLGGNH SRIEGPYVLY NPDTQYYYLY
LSYGGLDATG GYNIRVARSK KPDGPYYDAE GNPMLDVRGK GGTFFDDRSI EPYGVKLMGS
YTFETENEKG TGYVSPGHNS AYYDEKTGRS YLIFHTRFPG RGEEHEVRVH QLFMNKDGWP
VAAPYRYAGE TLKEVKQKDI TGTYKLIQHG KDISADIKQT INIQLNKNHT ISGEMTGTWR
KTGKNTADIT LAGKKYNGVF LRQWDSVREK NVMTFSVLNT SGEAVWGSK
