INUS_LACJO
ID INUS_LACJO Reviewed; 797 AA.
AC Q74K42;
DT 26-NOV-2014, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 113.
DE RecName: Full=Inulosucrase {ECO:0000303|PubMed:18408060};
DE Short=IS {ECO:0000250|UniProtKB:D3WYV9};
DE EC=2.4.1.9 {ECO:0000269|PubMed:18408060, ECO:0000269|PubMed:21801732};
DE Flags: Precursor;
GN Name=inuJ {ECO:0000303|PubMed:18408060};
GN Synonyms=ftf {ECO:0000303|PubMed:18408060}; OrderedLocusNames=LJ_0913;
OS Lactobacillus johnsonii (strain CNCM I-12250 / La1 / NCC 533).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae;
OC Lactobacillus.
OX NCBI_TaxID=257314;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=CNCM I-1225 / La1 / NCC 533;
RX PubMed=14983040; DOI=10.1073/pnas.0307327101;
RA Pridmore R.D., Berger B., Desiere F., Vilanova D., Barretto C.,
RA Pittet A.-C., Zwahlen M.-C., Rouvet M., Altermann E., Barrangou R.,
RA Mollet B., Mercenier A., Klaenhammer T., Arigoni F., Schell M.A.;
RT "The genome sequence of the probiotic intestinal bacterium Lactobacillus
RT johnsonii NCC 533.";
RL Proc. Natl. Acad. Sci. U.S.A. 101:2512-2517(2004).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=CNCM I-1225 / La1 / NCC 533;
RX PubMed=18408060; DOI=10.1128/aem.00377-08;
RA Anwar M.A., Kralj S., van der Maarel M.J., Dijkhuizen L.;
RT "The probiotic Lactobacillus johnsonii NCC 533 produces high-molecular-mass
RT inulin from sucrose by using an inulosucrase enzyme.";
RL Appl. Environ. Microbiol. 74:3426-3433(2008).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) OF 145-708 OF APOENZYME AND COMPLEX
RP WITH THE TRANSFRUCTOSYLATION PRODUCT 1-KESTOSE AND CALCIUM AND MUTANT
RP ASN-272 IN COMPLEX WITH SUCROSE, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE,
RP AND MUTAGENESIS OF ASP-272; ASN-301 AND ASN-305.
RC STRAIN=CNCM I-1225 / La1 / NCC 533;
RX PubMed=21801732; DOI=10.1016/j.jmb.2011.07.031;
RA Pijning T., Anwar M.A., Boger M., Dobruchowska J.M., Leemhuis H., Kralj S.,
RA Dijkhuizen L., Dijkstra B.W.;
RT "Crystal structure of inulosucrase from Lactobacillus: insights into the
RT substrate specificity and product specificity of GH68 fructansucrases.";
RL J. Mol. Biol. 412:80-93(2011).
CC -!- FUNCTION: Fructosyltransferase that catalyzes the polymerization of the
CC fructose moiety of sucrose to produce inulin polymer and inulin
CC oligosaccharides such as 1-kestose and nystose.
CC {ECO:0000269|PubMed:18408060, ECO:0000269|PubMed:21801732}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[(2->1)-beta-D-fructosyl](n) + sucrose = [(2->1)-beta-D-
CC fructosyl](n+1) + D-glucose; Xref=Rhea:RHEA:15745, Rhea:RHEA-
CC COMP:10036, Rhea:RHEA-COMP:14296, ChEBI:CHEBI:4167,
CC ChEBI:CHEBI:17992, ChEBI:CHEBI:29084; EC=2.4.1.9;
CC Evidence={ECO:0000269|PubMed:18408060, ECO:0000269|PubMed:21801732};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:18408060};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 7. More than 85% of the activity is retained in the pH
CC range 4.5-6.0. Activity decreases sharply at pH 7.5.
CC {ECO:0000269|PubMed:18408060};
CC Temperature dependence:
CC Optimum temperature is 55 degrees Celsius. Activity drastically
CC decreases at 60 degrees Celsius. {ECO:0000269|PubMed:18408060};
CC -!- SUBCELLULAR LOCATION: Secreted, cell wall {ECO:0000255|PROSITE-
CC ProRule:PRU00477}; Peptidoglycan-anchor {ECO:0000255|PROSITE-
CC ProRule:PRU00477}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 68 family. {ECO:0000305}.
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DR EMBL; AE017198; AAS08734.1; -; Genomic_DNA.
DR RefSeq; WP_011161805.1; NC_005362.1.
DR PDB; 2YFR; X-ray; 1.75 A; A=145-708.
DR PDB; 2YFS; X-ray; 2.60 A; A=145-708.
DR PDB; 2YFT; X-ray; 1.85 A; A=145-708.
DR PDBsum; 2YFR; -.
DR PDBsum; 2YFS; -.
DR PDBsum; 2YFT; -.
DR AlphaFoldDB; Q74K42; -.
DR SMR; Q74K42; -.
DR STRING; 257314.LJ_0913; -.
DR CAZy; GH68; Glycoside Hydrolase Family 68.
DR PRIDE; Q74K42; -.
DR EnsemblBacteria; AAS08734; AAS08734; LJ_0913.
DR KEGG; ljo:LJ_0913; -.
DR eggNOG; COG1621; Bacteria.
DR HOGENOM; CLU_013854_0_0_9; -.
DR OMA; IWDSWPV; -.
DR BRENDA; 2.4.1.9; 2873.
DR SABIO-RK; Q74K42; -.
DR Proteomes; UP000000581; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-KW.
DR GO; GO:0047725; F:inulosucrase activity; IEA:UniProtKB-EC.
DR GO; GO:0050053; F:levansucrase activity; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0009758; P:carbohydrate utilization; IEA:InterPro.
DR CDD; cd08997; GH68; 1.
DR Gene3D; 2.115.10.20; -; 1.
DR InterPro; IPR003469; Glyco_hydro_68.
DR InterPro; IPR023296; Glyco_hydro_beta-prop_sf.
DR InterPro; IPR019931; LPXTG_anchor.
DR Pfam; PF02435; Glyco_hydro_68; 1.
DR Pfam; PF00746; Gram_pos_anchor; 1.
DR SUPFAM; SSF75005; SSF75005; 1.
DR PROSITE; PS50847; GRAM_POS_ANCHORING; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Cell wall;
KW Glycosyltransferase; Metal-binding; Peptidoglycan-anchor;
KW Reference proteome; Secreted; Signal; Transferase.
FT SIGNAL 1..36
FT /evidence="ECO:0000255"
FT CHAIN 37..764
FT /note="Inulosucrase"
FT /id="PRO_0000431248"
FT PROPEP 765..797
FT /note="Removed by sortase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00477"
FT /id="PRO_0000431249"
FT REGION 54..176
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 708..766
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT MOTIF 761..765
FT /note="LPXTG sorting signal"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00477"
FT COMPBIAS 65..92
FT /note="Basic and acidic residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 93..136
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 137..171
FT /note="Basic and acidic residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 710..748
FT /note="Pro residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 750..764
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 272
FT /note="Nucleophile"
FT /evidence="ECO:0000303|PubMed:21801732"
FT ACT_SITE 524
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000303|PubMed:21801732"
FT BINDING 271
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21801732"
FT BINDING 317
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFS"
FT BINDING 340
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21801732"
FT BINDING 419
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFR, ECO:0007744|PDB:2YFS,
FT ECO:0007744|PDB:2YFT"
FT BINDING 424..425
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21801732"
FT BINDING 450
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFR, ECO:0007744|PDB:2YFS,
FT ECO:0007744|PDB:2YFT"
FT BINDING 487
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFR, ECO:0007744|PDB:2YFS,
FT ECO:0007744|PDB:2YFT"
FT BINDING 489
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFR, ECO:0007744|PDB:2YFS,
FT ECO:0007744|PDB:2YFT"
FT BINDING 521
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFR, ECO:0007744|PDB:2YFS,
FT ECO:0007744|PDB:2YFT"
FT BINDING 522..524
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21801732"
FT BINDING 542
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21801732"
FT BINDING 660
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFS"
FT BINDING 662
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFS"
FT BINDING 667
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:21801732,
FT ECO:0007744|PDB:2YFS"
FT SITE 425
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000303|PubMed:21801732"
FT MOD_RES 764
FT /note="Pentaglycyl murein peptidoglycan amidated alanine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00477"
FT MUTAGEN 272
FT /note="D->N: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:21801732"
FT MUTAGEN 301..303
FT /note="Missing: 1.5% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:21801732"
FT MUTAGEN 301
FT /note="N->A: 25% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:21801732"
FT MUTAGEN 301
FT /note="N->S: 40% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:21801732"
FT MUTAGEN 305
FT /note="N->A: 29% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:21801732"
FT MUTAGEN 305
FT /note="N->S: 50% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:21801732"
FT HELIX 181..189
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 194..196
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 199..205
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 215..219
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 222..234
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 237..239
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 246..248
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 253..255
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 256..258
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 260..262
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 263..267
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 269..277
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 279..281
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 290..298
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 306..313
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 319..321
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 323..327
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 336..345
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 351..359
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 361..364
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 367..380
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 381..383
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 385..398
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 402..405
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 408..414
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 424..430
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 436..444
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 452..456
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 458..460
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 465..477
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 479..487
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 490..497
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 501..503
FT /evidence="ECO:0007829|PDB:2YFT"
FT STRAND 506..509
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 513..515
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 517..519
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 524..531
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 534..542
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 543..545
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 549..559
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 563..575
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 580..583
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 584..588
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 599..607
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 610..621
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 624..627
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 637..644
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 645..647
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 648..658
FT /evidence="ECO:0007829|PDB:2YFR"
FT HELIX 665..667
FT /evidence="ECO:0007829|PDB:2YFS"
FT HELIX 670..672
FT /evidence="ECO:0007829|PDB:2YFR"
FT STRAND 678..680
FT /evidence="ECO:0007829|PDB:2YFS"
FT STRAND 683..685
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 687..691
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 695..697
FT /evidence="ECO:0007829|PDB:2YFR"
FT TURN 700..703
FT /evidence="ECO:0007829|PDB:2YFR"
SQ SEQUENCE 797 AA; 87186 MW; 3E1F6460C93342C8 CRC64;
MLENKNHKKI SLSGKSLLMG TLSTAAIVLS ASTANAATIN ADNVNENQTV EVTASSVNNE
NNKQVTEKDS ADKSTSDVAE DANTKKSNEN TETTEKNTQT VVTNAPVSDV KNTNTVTAET
PVDKVVNNSD QKTTNAATTD TKKDDVKQVE KKDSVDKTNA EENKDSSVKP AENATKAELK
GQVKDIVEES GVDTSKLTND QINELNKINF SKEAKSGTQL TYNDFKKIAK TLIEQDARYA
IPFFNASKIK NMPAAKTLDA QSGKVEDLEI WDSWPVQDAK TGYVSNWNGY QLVIGMMGVP
NVNDNHIYLL YNKYGDNDFN HWKNAGPIFG LGTPVIQQWS GSATLNKDGS IQLYYTKVDT
SDNNTNHQKL ASATVYLNLE KDQDKISIAH VDNDHIVFEG DGYHYQTYDQ WKETNKGADN
IAMRDAHVID DDNGNRYLVF EASTGTENYQ GDDQIYQWLN YGGTNKDNLG DFFQILSNSD
IKDRAKWSNA AIGIIKLNDD VKNPSVAKVY SPLISAPMVS DEIERPDVVK LGNKYYLFAA
TRLNRGSNDD AWMATNKAVG DNVAMIGYVS DNLTHGYVPL NESGVVLTAS VPANWRTATY
SYYAVPVEGR DDQLLITSYI TNRGEVAGKG MHATWAPSFL LQINPDNTTT VLAKMTNQGD
WIWDDSSENP DMMGVLEKDA PNSAALPGEW GKPVDWDLIG GYNLKPHQPV TPIPNVPTTP
ETPTTPDKPE VPTTPEVPTT PETPTPEAPK NPVKKTSQSK LPKAGDKNSF AAVVLGAVSS
ILGAVGLTGV SKRKRNN
