GGAP_MEISD
ID GGAP_MEISD Reviewed; 555 AA.
AC D7BAR0;
DT 22-NOV-2017, integrated into UniProtKB/Swiss-Prot.
DT 10-AUG-2010, sequence version 1.
DT 25-MAY-2022, entry version 59.
DE RecName: Full=Glucosylglycerate phosphorylase {ECO:0000303|PubMed:28754708};
DE Short=GGa phosphorylase {ECO:0000303|PubMed:28754708};
DE Short=GGaP {ECO:0000303|PubMed:28754708};
DE EC=2.4.1.352 {ECO:0000269|PubMed:28754708};
GN OrderedLocusNames=Mesil_0665 {ECO:0000312|EMBL:ADH62582.1};
OS Meiothermus silvanus (strain ATCC 700542 / DSM 9946 / VI-R2) (Thermus
OS silvanus).
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae;
OC Meiothermus.
OX NCBI_TaxID=526227;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700542 / DSM 9946 / VI-R2;
RX PubMed=21304690; DOI=10.4056/sigs.1042812;
RA Sikorski J., Tindall B.J., Lowry S., Lucas S., Nolan M., Copeland A.,
RA Glavina Del Rio T., Tice H., Cheng J.F., Han C., Pitluck S., Liolios K.,
RA Ivanova N., Mavromatis K., Mikhailova N., Pati A., Goodwin L., Chen A.,
RA Palaniappan K., Land M., Hauser L., Chang Y.J., Jeffries C.D., Rohde M.,
RA Goker M., Woyke T., Bristow J., Eisen J.A., Markowitz V., Hugenholtz P.,
RA Kyrpides N.C., Klenk H.P., Lapidus A.;
RT "Complete genome sequence of Meiothermus silvanus type strain (VI-R2).";
RL Stand. Genomic Sci. 3:37-46(2010).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, 3D-STRUCTURE MODELING, MUTAGENESIS OF ASN-275 AND GLU-383, AND
RP BIOTECHNOLOGY.
RX PubMed=28754708; DOI=10.1128/aem.01434-17;
RA Franceus J., Pinel D., Desmet T.;
RT "Glucosylglycerate phosphorylase, an enzyme with novel specificity involved
RT in compatible solute metabolism.";
RL Appl. Environ. Microbiol. 83:0-0(2017).
CC -!- FUNCTION: Catalyzes the reversible phosphorolysis of glucosylglycerate
CC into alpha-D-glucose 1-phosphate (Glc1P) and D-glycerate. May be a
CC regulator of intracellular levels of glucosylglycerate, a compatible
CC solute that primarily protects organisms facing salt stress and very
CC specific nutritional constraints. Has a very strict substrate
CC specificity. Cannot catalyze the phosphorolysis of sucrose or
CC synthesize sucrose from Glc1P and D-fructose.
CC {ECO:0000269|PubMed:28754708}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R)-2-O-(alpha-D-glucopyranosyl)-glycerate + phosphate = (R)-
CC glycerate + alpha-D-glucose 1-phosphate; Xref=Rhea:RHEA:55268,
CC ChEBI:CHEBI:16659, ChEBI:CHEBI:43474, ChEBI:CHEBI:58601,
CC ChEBI:CHEBI:62510; EC=2.4.1.352;
CC Evidence={ECO:0000269|PubMed:28754708};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.5 mM for 2-O-(alpha-D-glucopyranosyl)-D-glycerate (at pH 6.0 and
CC 42 degrees Celsius) {ECO:0000269|PubMed:28754708};
CC KM=2.5 mM for phosphate (at pH 6.0 and 42 degrees Celsius)
CC {ECO:0000269|PubMed:28754708};
CC KM=8.1 mM for alpha-D-glucose 1-phosphate (at pH 6.5 and 42 degrees
CC Celsius) {ECO:0000269|PubMed:28754708};
CC KM=2.6 mM for D-glycerate (at pH 6.5 and 42 degrees Celsius)
CC {ECO:0000269|PubMed:28754708};
CC Note=kcat is 0.83 sec(-1) towards 2-O-(alpha-D-glucopyranosyl)-D-
CC glycerate for the phosphorolytic direction (at pH 6.0 and 42 degrees
CC Celsius). kcat is 0.98 sec(-1) towards phosphate for the
CC phosphorolytic direction (at pH 6.0 and 42 degrees Celsius). kcat is
CC 96 sec(-1) towards alpha-D-glucose 1-phosphate for the synthetic
CC direction (at pH 6.5 and 42 degrees Celsius). kcat is 128 sec(-1)
CC towards D-glycerate for the synthetic direction (at pH 6.5 and 42
CC degrees Celsius). {ECO:0000269|PubMed:28754708};
CC pH dependence:
CC Optimum pH is 6 and 6.5 in phosphorolytic and synthetic directions,
CC respectively. {ECO:0000269|PubMed:28754708};
CC Temperature dependence:
CC Optimum temperature is 42 degrees Celsius. Retains full activity
CC after incubating at up to 55 degrees Celsius for 10 minutes, but only
CC 35% is left after incubating at 60 degrees Celsius.
CC {ECO:0000269|PubMed:28754708};
CC -!- BIOTECHNOLOGY: The excellent specific activity of this enzyme towards
CC glycerate opens up new options for the commercial production of
CC glucosylglycerate. This compound has been reported to be an
CC exceptionally potent and versatile stabilizer of proteins at elevated
CC temperatures. {ECO:0000305|PubMed:28754708}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family.
CC Glucosylglycerate phosphorylase subfamily. {ECO:0000305}.
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DR EMBL; CP002042; ADH62582.1; -; Genomic_DNA.
DR RefSeq; WP_013157171.1; NC_014212.1.
DR AlphaFoldDB; D7BAR0; -.
DR SMR; D7BAR0; -.
DR STRING; 526227.Mesil_0665; -.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR EnsemblBacteria; ADH62582; ADH62582; Mesil_0665.
DR KEGG; msv:Mesil_0665; -.
DR eggNOG; COG0366; Bacteria.
DR HOGENOM; CLU_021358_0_0_0; -.
DR OMA; PNATQFT; -.
DR OrthoDB; 1573900at2; -.
DR BRENDA; 2.4.1.352; 15676.
DR Proteomes; UP000001916; Chromosome.
DR GO; GO:0016757; F:glycosyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW.
DR CDD; cd11356; AmyAc_Sucrose_phosphorylase-like_1; 1.
DR Gene3D; 3.90.400.10; -; 1.
DR InterPro; IPR033746; GGa_phosphorylase.
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 Pfam; PF00128; Alpha-amylase; 1.
DR PIRSF; PIRSF003059; Sucrose_phosphorylase; 1.
DR SMART; SM00642; Aamy; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
PE 1: Evidence at protein level;
KW Carbohydrate metabolism; Glycosyltransferase; Reference proteome;
KW Transferase.
FT CHAIN 1..555
FT /note="Glucosylglycerate phosphorylase"
FT /id="PRO_0000442433"
FT ACT_SITE 231
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:A0ZZH6"
FT MUTAGEN 275
FT /note="N->A: 75-fold decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:28754708"
FT MUTAGEN 275
FT /note="N->H: 38-fold decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:28754708"
FT MUTAGEN 383
FT /note="E->A,Q: More than 100-fold decrease in catalytic
FT activity."
FT /evidence="ECO:0000269|PubMed:28754708"
SQ SEQUENCE 555 AA; 61531 MW; 6EFA0A04A23A3E5B CRC64;
MSSLTPELRQ SILEHLGFLY GERAPAVLGR LEEICSGFPA QRREGGWSEK DALLITYGDQ
IHAEGEPPLQ TLYDFLYERL RGVFSGVHLL PFYPSTSDDG FSVVDFQRVD PELGTWTDIR
IIAQDFRLMA DLVCNHVSAS SPWFQGFLQD DPQYQGFFIT VDPGTDLSTV FRPRALPLLT
PFQTPSGEKL VWTTFSPDQT DLNYANPEVL LEVIEALLCY VRNGAGLIRL DAVGFIWKEI
GTSCMHLEGA HRIVKLMRLV LDAVAPHVLL VSETNAPHRE NISYFGNGHD EAQLVYQFPL
PPLVMHTFRT GDASKLAGWA AGLTLPSERT TFFNFLASHD GIGVVPAGGI LQPEEIAALV
RQALEHGGRV NHKDTPDGPV PYELCLTLFD ALSNPNSDEA EDLKIARFLA ANVILLSLQG
IPGVYIHSLF GSPSDHAGFE ESGIPRRLNR HKFTKAELEE RLADPASRAA KILAAYSHLL
RVRSMHPAFH PNAPQRILPS TEVLRIVRGE GDQAVGCYIN VTDRPQVVSR IGKNLITGQW
FTGVLKPYQA AWIID
