PLH30_FORAG
ID PLH30_FORAG Reviewed; 516 AA.
AC T2KNC2;
DT 16-OCT-2019, integrated into UniProtKB/Swiss-Prot.
DT 16-OCT-2019, sequence version 2.
DT 03-AUG-2022, entry version 25.
DE RecName: Full=Endo-acting ulvan lyase {ECO:0000305};
DE EC=4.2.2.- {ECO:0000269|PubMed:29948117, ECO:0000269|PubMed:31285597};
DE AltName: Full=Endolytic ulvan lyase {ECO:0000303|PubMed:29948117};
DE AltName: Full=Polysaccharide lyase 28 family protein P30 {ECO:0000305};
DE Short=P30_PL28 {ECO:0000303|PubMed:31285597};
DE AltName: Full=Polysaccharide utilization locus H protein P30 {ECO:0000303|PubMed:31285597};
DE Short=PUL H protein P30;
DE Flags: Precursor;
GN ORFNames=BN863_22190;
OS Formosa agariphila (strain DSM 15362 / KCTC 12365 / LMG 23005 / KMM 3901 /
OS M-2Alg 35-1).
OC Bacteria; Bacteroidetes; Flavobacteriia; Flavobacteriales;
OC Flavobacteriaceae; Formosa.
OX NCBI_TaxID=1347342;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 15362 / KCTC 12365 / LMG 23005 / KMM 3901 / M-2Alg 35-1;
RX PubMed=23995932; DOI=10.1128/aem.01937-13;
RA Mann A.J., Hahnke R.L., Huang S., Werner J., Xing P., Barbeyron T.,
RA Huettel B., Stueber K., Reinhardt R., Harder J., Gloeckner F.O.,
RA Amann R.I., Teeling H.;
RT "The genome of the alga-associated marine flavobacterium Formosa agariphila
RT KMM 3901T reveals a broad potential for degradation of algal
RT polysaccharides.";
RL Appl. Environ. Microbiol. 79:6813-6822(2013).
RN [2]
RP REVISION OF GENE MODEL, FUNCTION, AND SUBCELLULAR LOCATION.
RX DOI=10.1016/j.algal.2017.09.025;
RA Salinas A., French C.E.;
RT "The enzymatic ulvan depolymerisation system from the alga-associated
RT marine flavobacterium Formosa agariphila.";
RL Algal Res. 27:335-344(2017).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND
RP SUBCELLULAR LOCATION.
RC STRAIN=DSM 15362 / KCTC 12365 / LMG 23005 / KMM 3901 / M-2Alg 35-1;
RX PubMed=29948117; DOI=10.1007/s00253-018-9142-y;
RA Reisky L., Stanetty C., Mihovilovic M.D., Schweder T., Hehemann J.H.,
RA Bornscheuer U.T.;
RT "Biochemical characterization of an ulvan lyase from the marine
RT flavobacterium Formosa agariphila KMM 3901T.";
RL Appl. Microbiol. Biotechnol. 102:6987-6996(2018).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, AND INDUCTION.
RX PubMed=31285597; DOI=10.1038/s41589-019-0311-9;
RA Reisky L., Prechoux A., Zuehlke M.K., Baeumgen M., Robb C.S., Gerlach N.,
RA Roret T., Stanetty C., Larocque R., Michel G., Song T., Markert S.,
RA Unfried F., Mihovilovic M.D., Trautwein-Schult A., Becher D., Schweder T.,
RA Bornscheuer U.T., Hehemann J.H.;
RT "A marine bacterial enzymatic cascade degrades the algal polysaccharide
RT ulvan.";
RL Nat. Chem. Biol. 15:803-812(2019).
CC -!- FUNCTION: Ulvan lyase involved in ulvan degradation (Ref.2). Ulvan is
CC the main polysaccharide component of the Ulvales (green seaweed) cell
CC wall. It is composed of disaccharide building blocks comprising 3-
CC sulfated rhamnose (Rha3S) linked to D-glucuronic acid (GlcA), L-
CC iduronic acid (IduA), or D-xylose (Xyl) (Probable). Ulvan lyase
CC catalyzes the endolytic cleavage of the glycosidic bond between Rha3S
CC and the uronic acids GlcA or IduA, producing oligosaccharides that have
CC unsaturated 4-deoxy-L-threo-hex-4-enopyranosiduronic acid (deltaUA) at
CC the non-reducing end. This results eventually in the degradation of the
CC ulvan polysaccharide into deltaUA-Rha3S disaccharides and deltaUA-
CC Rha3S-Xyl-Rha3S tetrasaccharides (PubMed:29948117, PubMed:31285597).
CC {ECO:0000269|PubMed:29948117, ECO:0000269|PubMed:31285597,
CC ECO:0000269|Ref.2, ECO:0000305|PubMed:31285597, ECO:0000305|Ref.2}.
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:29948117};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.26 mg/ml for ulvan (at 100 mM NaCl)
CC {ECO:0000269|PubMed:29948117};
CC KM=0.75 mg/ml for ulvan (at 600 mM NaCl)
CC {ECO:0000269|PubMed:29948117};
CC Note=kcat is 24.8 sec(-1) with ulvan as substrate (at 100 mM NaCl)
CC and 20.1 sec(-1) with ulvan as substrate (at 600 mM NaCl).
CC {ECO:0000269|PubMed:29948117};
CC pH dependence:
CC Optimum pH is 8.5. {ECO:0000269|PubMed:29948117};
CC Temperature dependence:
CC Optimum temperature is 29.5 degrees Celsius.
CC {ECO:0000269|PubMed:29948117};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000305|PubMed:31285597}.
CC Note=Secreted via the type IX secretion system (T9SS).
CC {ECO:0000305|PubMed:29948117, ECO:0000305|Ref.2}.
CC -!- INDUCTION: By ulvan and rhamnose. {ECO:0000269|PubMed:31285597}.
CC -!- DOMAIN: The ulvan-binding domain binds strongly to ulvan, it does not
CC bind other polymers like alginate, heparin, dextran sulfate or iota
CC carrageenan. Presumably it serves to anchor the enzyme at the
CC substrate, thus improving catalysis. Notably, the catalytic domain
CC alone is more active than the full-length enzyme with the binding
CC domain attached. Possibly, the ulvan-binding domain helps the enzyme to
CC act on ulvan in its insoluble form embedded in the algal cell wall.
CC {ECO:0000250|UniProtKB:G8G2V6}.
CC -!- SIMILARITY: Belongs to the polysaccharide lyase 28 family.
CC {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CDF79931.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305|Ref.2};
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DR EMBL; HG315671; CDF79931.1; ALT_INIT; Genomic_DNA.
DR RefSeq; WP_038530530.1; NZ_HG315671.1.
DR AlphaFoldDB; T2KNC2; -.
DR SMR; T2KNC2; -.
DR STRING; 1347342.BN863_22190; -.
DR EnsemblBacteria; CDF79931; CDF79931; BN863_22190.
DR PATRIC; fig|1347342.6.peg.2226; -.
DR eggNOG; ENOG502ZBB6; Bacteria.
DR HOGENOM; CLU_544861_0_0_10; -.
DR Proteomes; UP000016160; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR InterPro; IPR035992; Ricin_B-like_lectins.
DR InterPro; IPR026444; Secre_tail.
DR Pfam; PF18962; Por_Secre_tail; 1.
DR SUPFAM; SSF50370; SSF50370; 1.
DR TIGRFAMs; TIGR04183; Por_Secre_tail; 1.
PE 1: Evidence at protein level;
KW Calcium; Disulfide bond; Lyase; Metal-binding; Reference proteome;
KW Secreted; Signal.
FT SIGNAL 1..24
FT /evidence="ECO:0000255"
FT CHAIN 25..429
FT /note="Endo-acting ulvan lyase"
FT /id="PRO_0000448327"
FT PROPEP 430..516
FT /note="Removed by the type IX secretion system (T9SS)"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:29948117"
FT /id="PRO_0000448328"
FT REGION 289..429
FT /note="Ulvan-binding domain"
FT /evidence="ECO:0000250|UniProtKB:G8G2V6"
FT ACT_SITE 143
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT ACT_SITE 260
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 42
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_note="structural"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 44
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_note="structural"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 62
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_note="structural"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 64
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_note="structural"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 67
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_note="structural"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 68
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_note="structural"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 138
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 191..195
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT BINDING 260..263
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT SITE 141
FT /note="Neutralizes the sugar carboxylate group at subsite
FT +1"
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
FT DISULFID 38..65
FT /evidence="ECO:0000250|UniProtKB:A0A084JZF2"
SQ SEQUENCE 516 AA; 56629 MW; D42CE0D2B8663350 CRC64;
MLEKTTLKNI ILIHFLMFLA VVTAQTAPDE DTSAITRCTA EGTNPVRETD IPNPVNVGTI
DDRSCYANYK ESTVYGKTWG VYNITFDSND FDTSLQPRIE RSLSRSSETG IGSYARLTGV
FRILEVGDTS GTSQDGTYLA QAKGKHTGGG GSPDPAICLY LAKPVYGTGE DADKQVSFDI
YAERILYRGG EGDGREIVFL KNVKKDEETN FELEVGFKED PNDVSKKIQY CNAVIGGDTF
NWNIPEPERG TESGIRYGAY RVKGGRAQIR WANTTYQKVE NVEVTNPGPI GDVYKLKNVA
TGQYLSDSGV SASAVIMSDS GEAQNNYWTF VESGSLFNID NETFGILRAP GAGGPGGAYV
VVSTTKEGPS SDGDKVWTIH YNESNDTYRF ESGSSGRFMY QEINGNVTHI SAMNTDDRSV
WKAIAVESLS VDENAILASD VRVFPNPASD SFTISLKTIN HVTVNIYDVL GNTIFKSEFN
GDTIQIRNKG QFKAGVYLIQ LTDKNNNKYH KKLIVK
