EALGL_STRMK
ID EALGL_STRMK Reviewed; 742 AA.
AC B2FSW8;
DT 11-MAY-2016, integrated into UniProtKB/Swiss-Prot.
DT 10-JUN-2008, sequence version 1.
DT 03-AUG-2022, entry version 66.
DE RecName: Full=Alginate lyase {ECO:0000305|PubMed:26913076};
DE EC=4.2.2.26 {ECO:0000305|PubMed:26913076};
DE AltName: Full=Exolytic alginate lyase {ECO:0000305};
DE AltName: Full=Exolytic polysaccharide lyase {ECO:0000303|PubMed:26913076};
DE Flags: Precursor;
GN OrderedLocusNames=Smlt2602 {ECO:0000312|EMBL:CAQ46078.1};
OS Stenotrophomonas maltophilia (strain K279a).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales;
OC Xanthomonadaceae; Stenotrophomonas; Stenotrophomonas maltophilia group.
OX NCBI_TaxID=522373;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K279a;
RX PubMed=18419807; DOI=10.1186/gb-2008-9-4-r74;
RA Crossman L.C., Gould V.C., Dow J.M., Vernikos G.S., Okazaki A.,
RA Sebaihia M., Saunders D., Arrowsmith C., Carver T., Peters N., Adlem E.,
RA Kerhornou A., Lord A., Murphy L., Seeger K., Squares R., Rutter S.,
RA Quail M.A., Rajandream M.A., Harris D., Churcher C., Bentley S.D.,
RA Parkhill J., Thomson N.R., Avison M.B.;
RT "The complete genome, comparative and functional analysis of
RT Stenotrophomonas maltophilia reveals an organism heavily shielded by drug
RT resistance determinants.";
RL Genome Biol. 9:R74.1-R74.13(2008).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, BIOTECHNOLOGY, MUTAGENESIS OF GLN-153; ASN-207; HIS-208;
RP TYR-264 AND TYR-455, 3D-STRUCTURE MODELING, ACTIVE SITES, AND REACTION
RP MECHANISM.
RC STRAIN=K279a;
RX PubMed=26913076; DOI=10.1186/s13068-016-0455-8;
RA MacDonald L.C., Weiler E.B., Berger B.W.;
RT "Engineering broad-spectrum digestion of polyuronides from an exolytic
RT polysaccharide lyase.";
RL Biotechnol. Biofuels 9:43-43(2016).
CC -!- FUNCTION: Polysaccharide lyase that catalyzes the depolymerization of
CC alginate via a beta-elimination mechanism, cleaving the beta-1,4
CC glycosidic bond between two adjacent sugar residues. Acts specifically
CC on alginate and each of its block structures, with highest activity
CC toward poly-beta-D-mannuronate (poly-ManA). Shows an exolytic mode of
CC action, producing unsaturated monomers. Displays a very low activity
CC against poly-beta-D-glucuronate (poly-GlcA), and is not active on poly-
CC alpha-D-galacturonate, hyaluronan, heparin, heparan sulfate and
CC chondroitin sulfate. {ECO:0000269|PubMed:26913076}.
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;
CC Evidence={ECO:0000305|PubMed:26913076};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000250|UniProtKB:Q21FJ0};
CC Note=The zinc ion likely plays a structural role.
CC {ECO:0000250|UniProtKB:Q21FJ0};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.67 mM for alginate {ECO:0000269|PubMed:26913076};
CC KM=0.41 mM for poly-ManA {ECO:0000269|PubMed:26913076};
CC KM=5.99 mM for poly-GulA {ECO:0000269|PubMed:26913076};
CC KM=0.57 mM for poly-MG {ECO:0000269|PubMed:26913076};
CC KM=1.40 mM for poly-GlcA {ECO:0000269|PubMed:26913076};
CC Note=kcat is 34.8 sec(-1) with alginate as substrate. kcat is 62.2
CC sec(-1) with poly-ManA as substrate. kcat is 34.6 sec(-1) with poly-
CC GulA as substrate. kcat is 22.2 sec(-1) with poly-MG as substrate.
CC kcat is 0.4 sec(-1) with poly-GlcA as substrate.
CC {ECO:0000269|PubMed:26913076};
CC pH dependence:
CC Optimum pH is 8.5. {ECO:0000269|PubMed:26913076};
CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:Q21FJ0}.
CC -!- SUBCELLULAR LOCATION: Periplasm {ECO:0000305}.
CC -!- BIOTECHNOLOGY: Is an attractive candidate for the broad-spectrum
CC digestion of polyuronides into fermentable monomers for biofuel
CC production. {ECO:0000305|PubMed:26913076}.
CC -!- SIMILARITY: Belongs to the polysaccharide lyase 17 family.
CC {ECO:0000305}.
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DR EMBL; AM743169; CAQ46078.1; -; Genomic_DNA.
DR RefSeq; WP_012480338.1; NC_010943.1.
DR AlphaFoldDB; B2FSW8; -.
DR SMR; B2FSW8; -.
DR STRING; 522373.Smlt2602; -.
DR CAZy; PL17; Polysaccharide Lyase Family 17.
DR PRIDE; B2FSW8; -.
DR EnsemblBacteria; CAQ46078; CAQ46078; Smlt2602.
DR KEGG; sml:Smlt2602; -.
DR PATRIC; fig|522373.3.peg.2443; -.
DR eggNOG; ENOG502Z7XC; Bacteria.
DR HOGENOM; CLU_022650_0_0_6; -.
DR OMA; EGPYYQR; -.
DR OrthoDB; 442785at2; -.
DR BRENDA; 4.2.2.26; 5134.
DR SABIO-RK; B2FSW8; -.
DR Proteomes; UP000008840; 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 Carbohydrate metabolism; Lyase; Metal-binding; Periplasm;
KW Polysaccharide degradation; Reference proteome; Signal; Zinc.
FT SIGNAL 1..26
FT /evidence="ECO:0000255"
FT CHAIN 27..742
FT /note="Alginate lyase"
FT /id="PRO_5002777618"
FT ACT_SITE 264
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:26913076"
FT ACT_SITE 418
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:26913076"
FT BINDING 143
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 153..156
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 204
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 208
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 263..266
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 420
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 438
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 443
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 469
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT BINDING 669
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q21FJ0"
FT SITE 207
FT /note="Neutralizes the sugar carboxylate group at subsite
FT +1"
FT /evidence="ECO:0000305|PubMed:26913076"
FT SITE 208
FT /note="Neutralizes the sugar carboxylate group at subsite
FT +1"
FT /evidence="ECO:0000305|PubMed:26913076"
FT MUTAGEN 153
FT /note="Q->A,N: 2- to 3-fold decrease in activity against
FT alginate and its three block structures."
FT /evidence="ECO:0000269|PubMed:26913076"
FT MUTAGEN 207
FT /note="N->L: Complete loss of activity against alginate and
FT its three block structures."
FT /evidence="ECO:0000269|PubMed:26913076"
FT MUTAGEN 208
FT /note="H->A: Displays same catalytic activity toward poly-
FT GlcA as wild-type."
FT /evidence="ECO:0000269|PubMed:26913076"
FT MUTAGEN 208
FT /note="H->F: Exhibits 30% of wild-type activity against
FT alginate and poly-ManA and 5 and 8% wild-type activity
FT against poly-GulA and poly-MG, respectively. Significantly
FT influences the substrate specificity since the mutant
FT exhibits significant exolytic activity toward poly-GlcA,
FT with a 35-fold increase in catalytic efficiency over wild-
FT type toward poly-GlcA, a non-alginate-based substrate."
FT /evidence="ECO:0000269|PubMed:26913076"
FT MUTAGEN 264
FT /note="Y->F: Complete loss of activity against alginate and
FT its three block structures."
FT /evidence="ECO:0000269|PubMed:26913076"
FT MUTAGEN 418
FT /note="H->F: Complete loss of activity against alginate and
FT its three block structures."
FT /evidence="ECO:0000269|PubMed:26913076"
FT MUTAGEN 455
FT /note="Y->F: Displays less than 1% alginate and poly-ManA
FT activity compared to wild-type, yet approximately 8 and 4%
FT wild-type activity toward poly-GulA and poly-MG."
FT /evidence="ECO:0000269|PubMed:26913076"
SQ SEQUENCE 742 AA; 82406 MW; 2E560A37F5F0A57B CRC64;
MRLQPLFVSL ALAAPCALLP TASLSAAPAA AARQADTAPV LVTAAQWQQM ASEGRRYPWF
AKEQARTEAT LKKMMKAGID VPVPRDKGGG RTHEQHKRNY QALLAAGTLY RLTGDRAYVD
YARDMLLQYA QLYPTLGPHP EGRGQIPGRV FWQVLNDSVW LVNAIQGYDA IRDALSAEDR
NTIESKVFRP MAEFLVSEPK NYDQIHNHAT WAVAATGMTG YVLRDQELVE KSLRGSQKDD
KFGFLRQIDL LFSPDGYYEE GPYYQRYALA PFLLFANAIE RNEPQRKIFA RRDGVLLKAV
DVLVQSSYGG LFFPINDAIL DKGIDTEELV AGIGIAYART GDDRLLSVAE QQKRLLLSPE
GLQVAQALAA NKAKPFDYHP MLLRDGPDGD RGGLAILRMN GERGQALVQK DTMQGMGHGH
FDKLNWLFYD NGNPVVTDYG AARFLNVEAK RGGIYLAENR SWAKQTVAHN TLVVDEQSHF
NGNWKRGEAH APQVRFFQAD ADTQIASATM RDAYPGVAFT RTQALLRHPD LGLPVVLDLL
QVHGDKAARY DLPLHFNGHI VTTGFEAEHF PSQRPVLGKD NGYQHLWLDA RSKPGSEPRS
LAWLLDGRFY TYRFGSSAPA QALLVESGAN DPEFNLRREP ALLQRVDGQK DVTFFSVLEP
HGEYNGTAEY VHGADSRIRE IVRTRGSDAE VIELRLASGA RIALGVADNS ATTSEHSVTV
DGHVYRWNGS HARLDRSKGD GK
