GGOP_MARAH
ID GGOP_MARAH Reviewed; 480 AA.
AC E4PMA5;
DT 23-MAY-2018, integrated into UniProtKB/Swiss-Prot.
DT 08-FEB-2011, sequence version 1.
DT 03-AUG-2022, entry version 63.
DE RecName: Full=Glucosylglycerol phosphorylase {ECO:0000303|PubMed:29470619};
DE Short=GGoP {ECO:0000303|PubMed:29470619};
DE EC=2.4.1.359 {ECO:0000269|PubMed:29470619};
DE AltName: Full=2-O-alpha-D-glucopyranosylglycerol:phosphate alpha-D-glucosyltransferase {ECO:0000303|PubMed:29470619};
DE AltName: Full=2-O-alpha-D-glucosylglycerol phosphorylase (retaining) {ECO:0000303|PubMed:29470619};
GN Name=gtfA {ECO:0000312|EMBL:ADP98617.1};
GN OrderedLocusNames=HP15_2853 {ECO:0000312|EMBL:ADP98617.1};
OS Marinobacter adhaerens (strain DSM 23420 / HP15).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Marinobacteraceae; Marinobacter.
OX NCBI_TaxID=225937;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 23420 / HP15;
RA Gaerdes A.A.M., Kaeppel E., Shezad A., Seebah S., Teeling H., Yarza P.,
RA Gloeckner F.O., Ullrich M.S.;
RT "Complete genome sequence of Marinobacter adhaerens type strain (HP15).";
RL Submitted (FEB-2010) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, 3D-STRUCTURE MODELING, MUTAGENESIS OF TYR-194; ALA-333 AND
RP GLN-336, AND BIOTECHNOLOGY.
RC STRAIN=DSM 23420 / HP15;
RX PubMed=29470619; DOI=10.1007/s00253-018-8856-1;
RA Franceus J., Decuyper L., D'hooghe M., Desmet T.;
RT "Exploring the sequence diversity in glycoside hydrolase family 13_18
RT reveals a novel glucosylglycerol phosphorylase.";
RL Appl. Microbiol. Biotechnol. 102:3183-3191(2018).
CC -!- FUNCTION: Catalyzes the reversible phosphorolysis of 2-O-alpha-D-
CC glucosylglycerol with retention of the anomeric configuration, forming
CC alpha-D-glucose 1-phosphate and glycerol. Has most likely a catabolic
CC role, either regulating the intracellular levels of glucosylglycerol,
CC which acts as a compatible solute, or degrading it when the
CC environmental conditions change. Cannot catalyze the phosphorolysis of
CC sucrose or glucosylglycerate. {ECO:0000269|PubMed:29470619}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-O-(alpha-D-glucopyranosyl)glycerol + phosphate = alpha-D-
CC glucose 1-phosphate + glycerol; Xref=Rhea:RHEA:56416,
CC ChEBI:CHEBI:17754, ChEBI:CHEBI:43474, ChEBI:CHEBI:58601,
CC ChEBI:CHEBI:82766; EC=2.4.1.359;
CC Evidence={ECO:0000269|PubMed:29470619};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.9 mM for 2-O-(alpha-D-glucopyranosyl)glycerol (at pH 6.5 and 37
CC degrees Celsius) {ECO:0000269|PubMed:29470619};
CC KM=10 mM for phosphate (at pH 6.5 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:29470619};
CC KM=2.5 mM for alpha-D-glucose 1-phosphate (at pH 6.5 and 37 degrees
CC Celsius) {ECO:0000269|PubMed:29470619};
CC KM=7.0 mM for glycerol (at pH 6.5 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:29470619};
CC Note=kcat is 14 sec(-1) towards 2-O-(alpha-D-glucopyranosyl)glycerol
CC for the phosphorolytic direction (at pH 6.5 and 37 degrees Celsius).
CC kcat is 14 sec(-1) towards phosphate for the phosphorolytic direction
CC (at pH 6.5 and 37 degrees Celsius). kcat is 36 sec(-1) towards alpha-
CC D-glucose 1-phosphate for the synthetic direction (at pH 6.5 and 37
CC degrees Celsius). kcat is 39 sec(-1) towards glycerol for the
CC synthetic direction (at pH 6.5 and 37 degrees Celsius).
CC {ECO:0000269|PubMed:29470619};
CC pH dependence:
CC Optimum pH is 6.5. {ECO:0000269|PubMed:29470619};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius. Full activity is retained
CC after incubating at up to 45 degrees Celsius for 10 minutes, while
CC only 46% is left after incubating at 50 degrees Celsius. The protein
CC is completely denatured after incubating at higher temperatures.
CC {ECO:0000269|PubMed:29470619};
CC -!- BIOTECHNOLOGY: Might be an attractive biocatalyst for the production of
CC the osmolyte glucosylglycerol, which is currently produced on
CC industrial scale by exploiting a side activity of the closely related
CC sucrose phosphorylase. GGoP from M.adhaerens may offer far higher
CC productivities due to its high specific activity in the synthesis
CC direction. Glucosylglycerol is commercially distributed as a
CC moisturising agent in cosmetics and can also serve as a stabilizing
CC agent in protein formulations, or even as a functional food.
CC {ECO:0000305|PubMed:29470619}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. Sucrose
CC phosphorylase subfamily. {ECO:0000305}.
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DR EMBL; CP001978; ADP98617.1; -; Genomic_DNA.
DR RefSeq; WP_014578108.1; NC_017506.1.
DR AlphaFoldDB; E4PMA5; -.
DR SMR; E4PMA5; -.
DR STRING; 225937.HP15_2853; -.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR EnsemblBacteria; ADP98617; ADP98617; HP15_2853.
DR KEGG; mad:HP15_2853; -.
DR PATRIC; fig|225937.3.peg.2880; -.
DR eggNOG; COG0366; Bacteria.
DR HOGENOM; CLU_021358_1_0_6; -.
DR OMA; CFMLEET; -.
DR BioCyc; MetaCyc:MON-20515; -.
DR BRENDA; 2.4.1.359; 16299.
DR Proteomes; UP000007077; Chromosome.
DR GO; GO:0004645; F:1,4-alpha-oligoglucan phosphorylase activity; IEA:InterPro.
DR GO; GO:0016757; F:glycosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0051472; P:glucosylglycerol metabolic process; IDA:UniProtKB.
DR CDD; cd11355; AmyAc_Sucrose_phosphorylase; 1.
DR Gene3D; 3.90.400.10; -; 1.
DR InterPro; IPR006047; Glyco_hydro_13_cat_dom.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR InterPro; IPR045857; O16G_dom_2.
DR InterPro; IPR016377; Sucrose_GGa_phosphorylase-rel.
DR InterPro; IPR022527; Sucrose_phospho.
DR Pfam; PF00128; Alpha-amylase; 1.
DR PIRSF; PIRSF003059; Sucrose_phosphorylase; 1.
DR SMART; SM00642; Aamy; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR TIGRFAMs; TIGR03852; sucrose_gtfA; 1.
PE 1: Evidence at protein level;
KW Carbohydrate metabolism; Glycosyltransferase; Transferase.
FT CHAIN 1..480
FT /note="Glucosylglycerol phosphorylase"
FT /id="PRO_0000444046"
FT ACT_SITE 190
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:A0ZZH6"
FT ACT_SITE 231
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:A0ZZH6"
FT BINDING 194
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:29470619"
FT BINDING 336
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:29470619"
FT MUTAGEN 194
FT /note="Y->A: 2.7% of wild-type catalytic activity."
FT /evidence="ECO:0000269|PubMed:29470619"
FT MUTAGEN 333
FT /note="A->D: 0.3% of wild-type catalytic activity. Does not
FT gain activity on sucrose."
FT /evidence="ECO:0000269|PubMed:29470619"
FT MUTAGEN 336
FT /note="Q->A: 2.6% of wild-type catalytic activity."
FT /evidence="ECO:0000269|PubMed:29470619"
SQ SEQUENCE 480 AA; 54825 MW; DC990A3BD05E0C9E CRC64;
MLLKNAVQLI CYPDRIGNNL KDLYTVVDTH LSEAIGGLHI LPFFPSNADG GFSPLTHKEV
DPKVGTWDDI EAFTAKYDLC VDLTVNHISD ESPEFTDFIA NGFDSEYADL FVHVDKFGEI
SPDDMAKIHI RKEKEPFREV TLSDGTKTRV WCTFTEQQID LNYESDLAYQ LMESYIGFLT
SKGVNLLRLD AFGYTTKRIG TSCFLVEPEV YQILDWVNQV ALKHGAECLP EVHDHTSYQY
AISRRNMHPY GFALPPLLLY SLLDANSTYL KNWLRMCPRN MVTVLDTHDG ICIPDVEGVL
PDEKIKVLID NIDARSADPI MRRSAANIHS VGAIYQLTCT FYDALMQNDD AYIAARAIQF
FTPGIPQVYY VGLLAGCNDH ELMEQSGELR DINRHYYTLE EVEQDIQKPV VQRLLSLMKF
RSNYPAFDGH FELNYSNNSS VAMAWRHGDY YCHLFVDLNF KTVKVTYTDV ETGETRHLEC
