EALGL_SACD2
ID EALGL_SACD2 Reviewed; 736 AA.
AC Q21FJ0;
DT 11-MAY-2016, integrated into UniProtKB/Swiss-Prot.
DT 18-APR-2006, sequence version 1.
DT 03-AUG-2022, entry version 83.
DE RecName: Full=Exo-oligoalginate lyase {ECO:0000303|PubMed:22281843};
DE EC=4.2.2.26 {ECO:0000269|PubMed:22281843, ECO:0000269|PubMed:24478312};
DE AltName: Full=Exo-type alginate lyase {ECO:0000303|PubMed:22281843};
DE AltName: Full=Exolytic alginate lyase {ECO:0000305};
DE Flags: Precursor;
GN Name=alg17C {ECO:0000303|PubMed:22281843};
GN Synonyms=alg17A {ECO:0000312|EMBL:ABD82539.1};
GN OrderedLocusNames=Sde_3284 {ECO:0000312|EMBL:ABD82539.1};
OS Saccharophagus degradans (strain 2-40 / ATCC 43961 / DSM 17024).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Cellvibrionales;
OC Cellvibrionaceae; Saccharophagus.
OX NCBI_TaxID=203122;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=2-40 / ATCC 43961 / DSM 17024;
RX PubMed=18516288; DOI=10.1371/journal.pgen.1000087;
RA Weiner R.M., Taylor L.E. II, Henrissat B., Hauser L., Land M.,
RA Coutinho P.M., Rancurel C., Saunders E.H., Longmire A.G., Zhang H.,
RA Bayer E.A., Gilbert H.J., Larimer F., Zhulin I.B., Ekborg N.A., Lamed R.,
RA Richardson P.M., Borovok I., Hutcheson S.;
RT "Complete genome sequence of the complex carbohydrate-degrading marine
RT bacterium, Saccharophagus degradans strain 2-40 T.";
RL PLoS Genet. 4:E1000087-E1000087(2008).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP BIOTECHNOLOGY.
RC STRAIN=2-40 / ATCC 43961 / DSM 17024;
RX PubMed=22281843; DOI=10.1007/s00253-012-3882-x;
RA Kim H.T., Chung J.H., Wang D., Lee J., Woo H.C., Choi I.G., Kim K.H.;
RT "Depolymerization of alginate into a monomeric sugar acid using Alg17C, an
RT exo-oligoalginate lyase cloned from Saccharophagus degradans 2-40.";
RL Appl. Microbiol. Biotechnol. 93:2233-2239(2012).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF 24-736 OF WILD-TYPE AND MUTANTS
RP LEU-202 AND ALA-258 IN COMPLEX WITH ZINC AND A TRISACCHARIDE SUBSTRATE,
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP SUBUNIT, MUTAGENESIS OF ASN-149; ASN-201; HIS-202; TYR-258; ARG-260;
RP HIS-415; ARG-438 AND TYR-450, REACTION MECHANISM, AND ACTIVE SITE.
RC STRAIN=2-40 / ATCC 43961 / DSM 17024;
RX PubMed=24478312; DOI=10.1074/jbc.m113.531111;
RA Park D., Jagtap S., Nair S.K.;
RT "Structure of a PL17 family alginate lyase demonstrates functional
RT similarities among exotype depolymerases.";
RL J. Biol. Chem. 289:8645-8655(2014).
CC -!- FUNCTION: Catalyzes the depolymerization of alginate through an
CC exolytic mode of action, via a beta-elimination mechanism.
CC Preferentially acts on oligoalginates with degrees of polymerization
CC higher than 2 to produce the alginate monomer, 4-deoxy-L-erythro-5-
CC hexoseulose uronic acid. {ECO:0000269|PubMed:22281843,
CC ECO:0000269|PubMed:24478312}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Cleavage of 4-deoxy-alpha-L-erythro-hex-4-enopyranuronoside
CC oligosaccharides into 4-deoxy-alpha-L-erythro-hex-4-enopyranuronate
CC monosaccharides.; EC=4.2.2.26; Evidence={ECO:0000269|PubMed:22281843,
CC ECO:0000269|PubMed:24478312};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:24478312};
CC Note=The zinc ion seen in the crystal structure is located far away
CC from the active site and likely plays a structural role. However, it
CC seems important for enzyme activity since the mutation of one of its
CC coordination residues causes a high decrease in activity.
CC {ECO:0000269|PubMed:24478312};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=35.2 mg/ml for alginate (at pH 6 and 40 degrees Celsius)
CC {ECO:0000269|PubMed:22281843};
CC KM=22.3 uM for low viscosity alginate (at pH 7.5 and 30 degrees
CC Celsius) {ECO:0000269|PubMed:24478312};
CC KM=7.7 uM for alginate trisaccharide (at pH 7.5 and 30 degrees
CC Celsius) {ECO:0000269|PubMed:24478312};
CC Vmax=41.7 umol/min/mg enzyme with alginate as substrate (at pH 6 and
CC 40 degrees Celsius) {ECO:0000269|PubMed:22281843};
CC Note=kcat is 56.9 sec(-1) with low viscosity alginate as substrate.
CC kcat is 62.4 sec(-1) with alginate trisaccharide as substrate (at pH
CC 7.5 and 30 degrees Celsius). {ECO:0000269|PubMed:24478312};
CC pH dependence:
CC Optimum pH is 6. {ECO:0000269|PubMed:22281843};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:22281843};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:24478312}.
CC -!- SUBCELLULAR LOCATION: Periplasm {ECO:0000305}.
CC -!- BIOTECHNOLOGY: Could be used as the key enzyme to produce alginate
CC monomers in the process of utilizing alginate for biofuels and
CC chemicals production. {ECO:0000305|PubMed:22281843}.
CC -!- SIMILARITY: Belongs to the polysaccharide lyase 17 family.
CC {ECO:0000305}.
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DR EMBL; CP000282; ABD82539.1; -; Genomic_DNA.
DR RefSeq; WP_011469755.1; NC_007912.1.
DR PDB; 4NEI; X-ray; 1.85 A; A/B=1-736.
DR PDB; 4OJZ; X-ray; 1.90 A; A/B=1-736.
DR PDB; 4OK2; X-ray; 2.45 A; A/B=1-736.
DR PDB; 4OK4; X-ray; 1.70 A; A/B=1-736.
DR PDBsum; 4NEI; -.
DR PDBsum; 4OJZ; -.
DR PDBsum; 4OK2; -.
DR PDBsum; 4OK4; -.
DR AlphaFoldDB; Q21FJ0; -.
DR SMR; Q21FJ0; -.
DR STRING; 203122.Sde_3284; -.
DR CAZy; PL17; Polysaccharide Lyase Family 17.
DR PRIDE; Q21FJ0; -.
DR EnsemblBacteria; ABD82539; ABD82539; Sde_3284.
DR KEGG; sde:Sde_3284; -.
DR eggNOG; ENOG502Z7XC; Bacteria.
DR HOGENOM; CLU_022650_0_0_6; -.
DR OMA; EGPYYQR; -.
DR OrthoDB; 442785at2; -.
DR BRENDA; 4.2.2.11; 7555.
DR BRENDA; 4.2.2.3; 7555.
DR Proteomes; UP000001947; Chromosome.
DR GO; GO:0042597; C:periplasmic space; IEA:UniProtKB-SubCell.
DR GO; GO:0052764; F:exo-oligoalginate lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.50.10.100; -; 1.
DR InterPro; IPR008397; Alginate_lyase_dom.
DR InterPro; IPR008929; Chondroitin_lyas.
DR InterPro; IPR012480; Hepar_II_III.
DR Pfam; PF05426; Alginate_lyase; 1.
DR Pfam; PF07940; Hepar_II_III; 1.
DR SUPFAM; SSF48230; SSF48230; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Lyase; Metal-binding; Periplasm;
KW Polysaccharide degradation; Reference proteome; Signal; Zinc.
FT SIGNAL 1..23
FT /evidence="ECO:0000255"
FT CHAIN 24..736
FT /note="Exo-oligoalginate lyase"
FT /id="PRO_5004200187"
FT ACT_SITE 258
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:24478312"
FT ACT_SITE 413
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:B2FSW8"
FT BINDING 136
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT BINDING 146..149
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT BINDING 198
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT BINDING 202
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT BINDING 257..260
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT BINDING 415
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4NEI, ECO:0007744|PDB:4OJZ,
FT ECO:0007744|PDB:4OK2, ECO:0007744|PDB:4OK4"
FT BINDING 433
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4NEI, ECO:0007744|PDB:4OJZ,
FT ECO:0007744|PDB:4OK2, ECO:0007744|PDB:4OK4"
FT BINDING 438
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT BINDING 464
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4NEI, ECO:0007744|PDB:4OJZ,
FT ECO:0007744|PDB:4OK2, ECO:0007744|PDB:4OK4"
FT BINDING 667
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24478312,
FT ECO:0007744|PDB:4OJZ"
FT SITE 201
FT /note="Neutralizes the sugar carboxylate group at subsite
FT +1"
FT /evidence="ECO:0000305|PubMed:24478312"
FT SITE 202
FT /note="Neutralizes the sugar carboxylate group at subsite
FT +1"
FT /evidence="ECO:0000305|PubMed:24478312"
FT MUTAGEN 149
FT /note="N->A: 1600-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 201
FT /note="N->A: Complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 202
FT /note="H->L: 20-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 258
FT /note="Y->A: Complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 260
FT /note="R->A: 2000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 415
FT /note="H->A: 1000-fold decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 438
FT /note="R->A: 4400-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:24478312"
FT MUTAGEN 450
FT /note="Y->A: Complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:24478312"
FT STRAND 32..36
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 37..46
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 47..49
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 51..68
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 84..106
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 109..125
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 126..128
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 137..140
FT /evidence="ECO:0007829|PDB:4OJZ"
FT STRAND 142..146
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 147..162
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 165..167
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 170..179
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 181..188
FT /evidence="ECO:0007829|PDB:4OK4"
FT TURN 189..191
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 193..196
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 201..217
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 220..228
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 232..237
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 238..244
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 256..276
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 278..280
FT /evidence="ECO:0007829|PDB:4OK4"
FT TURN 282..284
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 285..299
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 321..334
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 339..346
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 353..363
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 375..378
FT /evidence="ECO:0007829|PDB:4OK4"
FT TURN 380..383
FT /evidence="ECO:0007829|PDB:4OJZ"
FT STRAND 385..396
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 400..407
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 411..413
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 420..425
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 428..431
FT /evidence="ECO:0007829|PDB:4OK4"
FT TURN 443..445
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 446..448
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 452..456
FT /evidence="ECO:0007829|PDB:4OK4"
FT TURN 457..459
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 461..463
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 464..466
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 474..477
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 479..483
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 488..494
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 496..505
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 507..509
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 512..521
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 526..528
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 530..551
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 553..560
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 568..572
FT /evidence="ECO:0007829|PDB:4OK4"
FT HELIX 578..580
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 581..588
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 594..600
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 603..612
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 618..625
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 638..659
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 661..663
FT /evidence="ECO:0007829|PDB:4OK4"
FT TURN 664..667
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 668..670
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 675..684
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 687..694
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 699..704
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 713..717
FT /evidence="ECO:0007829|PDB:4OK4"
FT STRAND 719..732
FT /evidence="ECO:0007829|PDB:4OK4"
SQ SEQUENCE 736 AA; 81582 MW; 500D22ED19B8DB80 CRC64;
MLSVNTIKNT LLAAVLVSVP ATAQVSGNGH PNLIVTEQDV ANIAASWESY DAYAEQLNAD
KTNLDAFMAE GVVVPMPKDA GGGYTHEQHK RNYKAIRNAG FLYQVTGDEK YLTFAKDLLL
AYAKMYPSLG EHPNRKEQSP GRLFWQSLNE AVWLVYSIQG YDAIIDGLAA EEKQEIESGV
FLPMAKFLSV ESPETFNKIH NHGTWAVAAV GMTGYVLGND ELVEISLMGL DKTGKAGFMK
QLDKLFSPDG YYTEGPYYQR YALMPFIWFA KAIETNEPER KIFEYRNNIL LKAVYTTIDL
SYAGYFFPIN DALKDKGIDT VELVHALAIV YSITGDNTLL DIAQEQGRIS LTGDGLKVAK
AVGEGLTQPY NYRSILLGDG ADGDQGALSI HRLGEGHNHM ALVAKNTSQG MGHGHFDKLN
WLLYDNGNEI VTDYGAARYL NVEAKYGGHY LAENNTWAKQ TIAHNTLVVN EQSHFYGDVT
TADLHHPEVL SFYSGEDYQL SSAKEANAYD GVEFVRSMLL VNVPSLEHPI VVDVLNVSAD
KASTFDLPLY FNGQIIDFSF KVKDNKNVMK MLGKRNGYQH LWLRNTAPVG DASERATWIL
DDRFYSYAFV TSTPSKKQNV LIAELGANDP NYNLRQQQVL IRRVEKAKQA SFVSVLEPHG
KYDGSLETTS GAYSNVKSVK HVSENGKDVV VVDLKDGSNV VVALSYNANS EQVHKVNAGE
EAIEWKGFSS VVVRRK
