ID   SACC_BACSU              Reviewed;         677 AA.
AC   P05656;
DT   01-NOV-1988, integrated into UniProtKB/Swiss-Prot.
DT   01-NOV-1988, sequence version 1.
DT   27-MAR-2024, entry version 161.
DE   RecName: Full=Levanase;
DE            EC=3.2.1.80;
DE   AltName: Full=Beta-D-fructofuranosidase;
DE   AltName: Full=Exo-beta-D-fructosidase;
DE   AltName: Full=Exo-levanase;
DE   Flags: Precursor;
GN   Name=sacC; OrderedLocusNames=BSU27030;
OS   Bacillus subtilis (strain 168).
OC   Bacteria; Bacillota; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX   NCBI_TaxID=224308;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX   PubMed=3112519; DOI=10.1007/bf00330439;
RA   Martin I., Debarbouille M., Ferrari E., Klier A., Rapoport G.;
RT   "Characterization of the levanase gene of Bacillus subtilis which shows
RT   homology to yeast invertase.";
RL   Mol. Gen. Genet. 208:177-184(1987).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=3120151; DOI=10.1093/nar/15.22.9606;
RA   Schoergendorfer K., Schwab H., Lafferty R.M.;
RT   "Nucleotide sequence of a cloned 2.5 kb PstI-EcoRI Bacillus subtilis DNA
RT   fragment coding for levanase.";
RL   Nucleic Acids Res. 15:9606-9606(1987).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9141695; DOI=10.1099/00221287-143-4-1321;
RA   Parro V., San Roman M., Galindo I., Purnelle B., Bolotin A., Sorokin A.,
RA   Mellado R.P.;
RT   "A 23911 bp region of the Bacillus subtilis genome comprising genes located
RT   upstream and downstream of the lev operon.";
RL   Microbiology 143:1321-1326(1997).
RN   [4]
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   [5]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-10.
RX   PubMed=2117666; DOI=10.1016/0022-2836(90)90284-s;
RA   Martin-Verstraete I., Debarbouille M., Klier A., Rapoport G.;
RT   "Levanase operon of Bacillus subtilis includes a fructose-specific
RT   phosphotransferase system regulating the expression of the operon.";
RL   J. Mol. Biol. 214:657-671(1990).
RN   [6]
RP   INDUCTION, AND TRANSCRIPTIONAL REGULATION.
RX   PubMed=2495266; DOI=10.1128/jb.171.4.1885-1892.1989;
RA   Martin I., Debarbouille M., Klier A., Rapoport G.;
RT   "Induction and metabolite regulation of levanase synthesis in Bacillus
RT   subtilis.";
RL   J. Bacteriol. 171:1885-1892(1989).
RN   [7]
RP   FUNCTION, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION, AND
RP   BIOPHYSICOCHEMICAL PROPERTIES.
RX   PubMed=7646030; DOI=10.1128/aem.61.5.1953-1958.1995;
RA   Wanker E., Huber A., Schwab H.;
RT   "Purification and characterization of the Bacillus subtilis levanase
RT   produced in Escherichia coli.";
RL   Appl. Environ. Microbiol. 61:1953-1958(1995).
RN   [8]
RP   SUBCELLULAR LOCATION, AND SECRETION PROCESS.
RX   PubMed=10217495; DOI=10.1099/13500872-145-3-613;
RA   Leloup L., Le Saux J., Petit-Glatron M.-F., Chambert R.;
RT   "Kinetics of the secretion of Bacillus subtilis levanase overproduced
RT   during the exponential phase of growth.";
RL   Microbiology 145:613-619(1999).
CC   -!- FUNCTION: Exo-fructosidase that can hydrolyze both levan and inulin,
CC       leading to the production of free fructose. Is also able to hydrolyze
CC       sucrose and to a small extent raffinose, but not melezitose,
CC       stachylose, cellobiose, maltose, and lactose.
CC       {ECO:0000269|PubMed:7646030}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Hydrolysis of terminal, non-reducing (2->1)- and (2->6)-linked
CC         beta-D-fructofuranose residues in fructans.; EC=3.2.1.80;
CC   -!- ACTIVITY REGULATION: Is completely inhibited by Ag(+) and Hg(2+) ions.
CC       {ECO:0000269|PubMed:7646030}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=1.2 uM for levan (at pH 5.5 and 55 degrees Celsius)
CC         {ECO:0000269|PubMed:7646030};
CC         KM=6.7 mM for inulin (at pH 5.5 and 55 degrees Celsius)
CC         {ECO:0000269|PubMed:7646030};
CC         KM=64 mM for sucrose (at pH 5.5 and 55 degrees Celsius)
CC         {ECO:0000269|PubMed:7646030};
CC       pH dependence:
CC         Optimum pH is 5.5 for inulin hydrolysis.
CC         {ECO:0000269|PubMed:7646030};
CC       Temperature dependence:
CC         Optimum temperature is 55 degrees Celsius for inulin hydrolysis. Is
CC         rapidly inactivated at 60 degrees Celsius. High stable at 50 and 55
CC         degrees Celsius. {ECO:0000269|PubMed:7646030};
CC   -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:10217495}.
CC   -!- INDUCTION: Induced by low concentrations of fructose, but not by
CC       sucrose. Repressed by glucose or high concentrations of fructose.
CC       {ECO:0000269|PubMed:2495266}.
CC   -!- MISCELLANEOUS: Levanase cannot be detected in the wild-type B.subtilis
CC       but is mostly secreted into the culture medium by SacL mutants,
CC       especially at the end of the exponential growth phase.
CC   -!- SIMILARITY: Belongs to the glycosyl hydrolase 32 family. {ECO:0000305}.
CC   -!- SEQUENCE CAUTION:
CC       Sequence=CAA29137.1; Type=Erroneous initiation; Evidence={ECO:0000305};
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DR   EMBL; X05649; CAA29137.1; ALT_INIT; Genomic_DNA.
DR   EMBL; Y00485; CAA68542.1; -; Genomic_DNA.
DR   EMBL; X92868; CAA63465.1; -; Genomic_DNA.
DR   EMBL; AL009126; CAB14645.1; -; Genomic_DNA.
DR   EMBL; X56098; CAA39581.1; -; Genomic_DNA.
DR   PIR; A27286; A27286.
DR   RefSeq; NP_390581.1; NC_000964.3.
DR   RefSeq; WP_004398804.1; NC_000964.3.
DR   PDB; 4AZZ; X-ray; 1.70 A; A/B=515-677.
DR   PDB; 4B1L; X-ray; 1.65 A; A=515-677.
DR   PDB; 4B1M; X-ray; 1.10 A; A/B/C=515-677.
DR   PDBsum; 4AZZ; -.
DR   PDBsum; 4B1L; -.
DR   PDBsum; 4B1M; -.
DR   AlphaFoldDB; P05656; -.
DR   SMR; P05656; -.
DR   STRING; 224308.BSU27030; -.
DR   CAZy; CBM66; Carbohydrate-Binding Module Family 66.
DR   CAZy; GH32; Glycoside Hydrolase Family 32.
DR   PaxDb; 224308-BSU27030; -.
DR   DNASU; 938092; -.
DR   EnsemblBacteria; CAB14645; CAB14645; BSU_27030.
DR   GeneID; 938092; -.
DR   KEGG; bsu:BSU27030; -.
DR   PATRIC; fig|224308.179.peg.2936; -.
DR   eggNOG; COG1621; Bacteria.
DR   InParanoid; P05656; -.
DR   OrthoDB; 9759709at2; -.
DR   PhylomeDB; P05656; -.
DR   BioCyc; BSUB:BSU27030-MONOMER; -.
DR   BRENDA; 3.2.1.80; 658.
DR   Proteomes; UP000001570; Chromosome.
DR   GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR   GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR   GO; GO:0051669; F:fructan beta-fructosidase activity; IEA:UniProtKB-EC.
DR   GO; GO:0004575; F:sucrose alpha-glucosidase activity; IBA:GO_Central.
DR   GO; GO:0005987; P:sucrose catabolic process; IBA:GO_Central.
DR   CDD; cd18622; GH32_Inu-like; 1.
DR   InterPro; IPR010496; 3-keto-disaccharide_hydrolase.
DR   InterPro; IPR013320; ConA-like_dom_sf.
DR   InterPro; IPR001362; Glyco_hydro_32.
DR   InterPro; IPR018053; Glyco_hydro_32_AS.
DR   InterPro; IPR013189; Glyco_hydro_32_C.
DR   InterPro; IPR013148; Glyco_hydro_32_N.
DR   InterPro; IPR023296; Glyco_hydro_beta-prop_sf.
DR   PANTHER; PTHR42800; EXOINULINASE INUD (AFU_ORTHOLOGUE AFUA_5G00480); 1.
DR   PANTHER; PTHR42800:SF1; EXOINULINASE INUD (AFU_ORTHOLOGUE AFUA_5G00480); 1.
DR   Pfam; PF06439; 3keto-disac_hyd; 1.
DR   Pfam; PF08244; Glyco_hydro_32C; 1.
DR   Pfam; PF00251; Glyco_hydro_32N; 1.
DR   SMART; SM00640; Glyco_32; 1.
DR   SUPFAM; SSF75005; Arabinanase/levansucrase/invertase; 1.
DR   SUPFAM; SSF49899; Concanavalin A-like lectins/glucanases; 1.
DR   PROSITE; PS00609; GLYCOSYL_HYDROL_F32; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Carbohydrate metabolism; Glycosidase; Hydrolase;
KW   Reference proteome; Secreted; Signal.
FT   SIGNAL          1..24
FT                   /evidence="ECO:0000255"
FT   CHAIN           25..677
FT                   /note="Levanase"
FT                   /id="PRO_0000033405"
FT   ACT_SITE        49
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU10067"
FT   BINDING         46..49
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         65
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         73
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         105..106
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         171..172
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         223
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         313
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   CONFLICT        658
FT                   /note="Q -> L (in Ref. 2; CAA68542)"
FT                   /evidence="ECO:0000305"
FT   STRAND          529..535
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          538..581
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          585..594
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   TURN            595..598
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          599..606
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          609..617
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          626..633
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          636..641
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          644..650
FT                   /evidence="ECO:0007829|PDB:4B1M"
FT   STRAND          657..676
FT                   /evidence="ECO:0007829|PDB:4B1M"
SQ   SEQUENCE   677 AA;  75951 MW;  80FD6B0A5EE7F525 CRC64;
     MKKRLIQVMI MFTLLLTMAF SADAADSSYY DEDYRPQYHF TPEANWMNDP NGMVYYAGEY
     HLFYQYHPYG LQWGPMHWGH AVSKDLVTWE HLPVALYPDE KGTIFSGSAV VDKNNTSGFQ
     TGKEKPLVAI YTQDREGHQV QSIAYSNDKG RTWTKYAGNP VIPNPGKKDF RDPKVFWYEK
     EKKWVMVLAA GDRILIYTSK NLKQWTYASE FGQDQGSHGG VWECPDLFEL PVDGNPNQKK
     WVMQVSVGNG AVSGGSGMQY FVGDFDGTHF KNENPPNKVL WTDYGRDFYA AVSWSDIPST
     DSRRLWLGWM SNWQYANDVP TSPWRSATSI PRELKLKAFT EGVRVVQTPV KELETIRGTS
     KKWKNLTISP ASHNVLAGQS GDAYEINAEF KVSPGSAAEF GFKVRTGENQ FTKVGYDRRN
     AKLFVDRSES GNDTFNPAFN TGKETAPLKP VNGKVKLRIF VDRSSVEVFG NDGKQVITDI
     ILPDRSSKGL ELYAANGGVK VKSLTIHPLK KVWGTTPFMS NMTGWTTVNG TWADTIEGKQ
     GRSDGDSFIL SSASGSDFTY ESDITIKDGN GRGAGALMFR SDKDAKNGYL ANVDAKHDLV
     KFFKFENGAA SVIAEYKTPI DVNKKYHLKT EAEGDRFKIY LDDRLVIDAH DSVFSEGQFG
     LNVWDATAVF QNVTKES
//