GLVA_BACSU
ID GLVA_BACSU Reviewed; 449 AA.
AC P54716;
DT 01-OCT-1996, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1996, sequence version 1.
DT 03-AUG-2022, entry version 151.
DE RecName: Full=Maltose-6'-phosphate glucosidase;
DE EC=3.2.1.122;
DE AltName: Full=6-phospho-alpha-D-glucosidase;
DE AltName: Full=6-phosphoryl-O-alpha-D-glucopyranosyl:phosphoglucohydrolase;
GN Name=glvA; Synonyms=glv-1, glvG, malA; OrderedLocusNames=BSU08180;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=8704981; DOI=10.1099/13500872-142-6-1417;
RA Yamamoto H., Uchiyama S., Fajar A.N., Ogasawara N., Sekiguchi J.;
RT "Determination of a 12 kb nucleotide sequence around the 76 degrees region
RT of the Bacillus subtilis chromosome.";
RL Microbiology 142:1417-1421(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [3]
RP PROTEIN SEQUENCE OF 1-37, CHARACTERIZATION, MASS SPECTROMETRY, AND
RP MUTAGENESIS.
RX PubMed=9765262; DOI=10.1074/jbc.273.42.27347;
RA Thompson J., Pikis A., Ruvinov S.B., Henrissat B., Yamamoto H.,
RA Sekiguchi J.;
RT "The gene glvA of Bacillus subtilis 168 encodes a metal-requiring, NAD(H)-
RT dependent 6-phospho-alpha-glucosidase. Assignment to family 4 of the
RT glycosylhydrolase superfamily.";
RL J. Biol. Chem. 273:27347-27356(1998).
RN [4]
RP INDUCTION.
RX PubMed=11489864; DOI=10.1128/jb.183.17.5110-5121.2001;
RA Yamamoto H., Serizawa M., Thompson J., Sekiguchi J.;
RT "Regulation of the glv operon in Bacillus subtilis: YfiA (GlvR) is a
RT positive regulator of the operon that is repressed through CcpA and cre.";
RL J. Bacteriol. 183:5110-5121(2001).
RN [5]
RP CRYSTALLIZATION.
RX PubMed=10329789; DOI=10.1107/s0907444999003790;
RA Varrot A., Yamamoto H., Sekiguchi J., Thompson J., Davies G.J.;
RT "Crystallization and preliminary X-ray analysis of the 6-phospho-alpha-
RT glucosidase from Bacillus subtilis.";
RL Acta Crystallogr. D 55:1212-1214(1999).
RN [6]
RP REACTION MECHANISM, CHARACTERIZATION, COFACTOR, SUBSTRATE SPECIFICITY,
RP ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=17676871; DOI=10.1021/bi700536p;
RA Yip V.L.Y., Thompson J., Withers S.G.;
RT "Mechanism of GlvA from Bacillus subtilis: a detailed kinetic analysis of a
RT 6-phospho-alpha-glucosidase from glycoside hydrolase family 4.";
RL Biochemistry 46:9840-9852(2007).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.05 ANGSTROMS) OF COMPLEX WITH NAD(H) AND
RP ALPHA-D-GLUCOSE-6-PHOSPHATE, AND REACTION MECHANISM.
RX PubMed=15341727; DOI=10.1016/j.str.2004.06.020;
RA Rajan S.S., Yang X., Collart F., Yip V.L.Y., Withers S.G., Varrot A.,
RA Thompson J., Davies G.J., Anderson W.F.;
RT "Novel catalytic mechanism of glycoside hydrolysis based on the structure
RT of an NAD+/Mn2+ -dependent phospho-alpha-glucosidase from Bacillus
RT subtilis.";
RL Structure 12:1619-1629(2004).
CC -!- FUNCTION: Hydrolyzes maltose-6'-phosphate and trehalose-6'-phosphate.
CC Is involved in the catabolism of alpha-glycosides accumulated via a
CC phosphoenolpyruvate-dependent maltose phosphotransferase system (PEP-
CC PTS). Is also able to significantly catalyze the hydrolysis of both 6-
CC phospho-alpha- and 6-phospho-beta-glucosides containing activated
CC leaving groups such as p-nitrophenol and does so with retention and
CC inversion, respectively, of the substrate anomeric configuration.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=alpha-maltose 6'-phosphate + H2O = D-glucose + D-glucose 6-
CC phosphate; Xref=Rhea:RHEA:20421, ChEBI:CHEBI:4167, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:57478, ChEBI:CHEBI:61548; EC=3.2.1.122;
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:17676871};
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:17676871};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:17676871};
CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC Evidence={ECO:0000269|PubMed:17676871};
CC Note=Binds 1 divalent metal cation per subunit. Manganese, iron, cobalt
CC or nickel enhance activity. {ECO:0000269|PubMed:17676871};
CC -!- COFACTOR:
CC Name=NAD(+); Xref=ChEBI:CHEBI:57540;
CC Evidence={ECO:0000269|PubMed:17676871};
CC Note=Binds 1 NAD(+) per subunit. Is only active with NAD(+), not NADH.
CC {ECO:0000269|PubMed:17676871};
CC -!- ACTIVITY REGULATION: Cellobiose-6'-phosphate and 6-phospho-beta-D-
CC glucopyranoside are not substrates but competitive inhibitors of GlvA.
CC {ECO:0000269|PubMed:17676871}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=400 uM for maltose-6'-phosphate (at pH 7.5 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:17676871};
CC KM=360 uM for maltose-6'-phosphate (at pH 8.4 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:17676871};
CC KM=610 uM for methyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37
CC degrees Celsius) {ECO:0000269|PubMed:17676871};
CC KM=69 uM for phenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37
CC degrees Celsius) {ECO:0000269|PubMed:17676871};
CC KM=52 uM for 4-nitrophenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and
CC 37 degrees Celsius) {ECO:0000269|PubMed:17676871};
CC KM=12 uM for 3,4-dinitrophenyl 6-phospho-alpha-D-glucoside (at pH 7.5
CC and 37 degrees Celsius) {ECO:0000269|PubMed:17676871};
CC KM=2.9 uM for 3,4-dinitrophenyl 6-phospho-beta-D-glucoside (at pH 7.5
CC and 37 degrees Celsius) {ECO:0000269|PubMed:17676871};
CC KM=27 uM for 3,5-dichlorophenyl 6-phospho-beta-D-glucoside (at pH 7.5
CC and 37 degrees Celsius) {ECO:0000269|PubMed:17676871};
CC KM=34 uM for 3-nitrophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and
CC 37 degrees Celsius) {ECO:0000269|PubMed:17676871};
CC pH dependence:
CC Optimum pH is about 7.6-8.4. Stable from pH 4.0 to 10.0.
CC {ECO:0000269|PubMed:17676871};
CC -!- SUBUNIT: Homotetramer.
CC -!- INDUCTION: By maltose; repressed by glucose.
CC {ECO:0000269|PubMed:11489864}.
CC -!- MASS SPECTROMETRY: Mass=50510; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:9765262};
CC -!- MISCELLANEOUS: Reaction proceeds via a redox-elimination-addition
CC mechanism consistent with an E1cb-type mechanism. This includes redox
CC steps involving NAD(+) and stabilization of intermediates by Mn(2+).
CC -!- MISCELLANEOUS: Because it hydrolyzes 6-phospho-alpha-glucopyranosides
CC without activated leaving groups (such as maltose-6'-phosphate) but not
CC glycosidic linkage of naturally occurring phospho-beta-glucosides such
CC as cellobiose-6'-phosphate nor methyl 6-phospho-beta-D-glucopyranoside,
CC GlvA is certainly more appropriately classified as a 6-phospho-alpha-
CC glucosidase than as a 6-phospho-beta-glucosidase. The ability to
CC hydrolyze beta-glycosidic linkages is exceptional and applies only to
CC activated 6-phospho-beta-D-glucopyranosides.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 4 family. {ECO:0000305}.
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DR EMBL; D50543; BAA09103.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB12647.1; -; Genomic_DNA.
DR PIR; F69635; F69635.
DR RefSeq; NP_388699.1; NC_000964.3.
DR RefSeq; WP_003244008.1; NZ_JNCM01000032.1.
DR PDB; 1U8X; X-ray; 2.05 A; X=1-449.
DR PDBsum; 1U8X; -.
DR AlphaFoldDB; P54716; -.
DR SMR; P54716; -.
DR STRING; 224308.BSU08180; -.
DR DrugBank; DB02007; alpha-D-glucose 6-phosphate.
DR CAZy; GH4; Glycoside Hydrolase Family 4.
DR PaxDb; P54716; -.
DR PRIDE; P54716; -.
DR DNASU; 936161; -.
DR EnsemblBacteria; CAB12647; CAB12647; BSU_08180.
DR GeneID; 936161; -.
DR KEGG; bsu:BSU08180; -.
DR PATRIC; fig|224308.179.peg.884; -.
DR eggNOG; COG1486; Bacteria.
DR OMA; METYSPD; -.
DR PhylomeDB; P54716; -.
DR BioCyc; BSUB:BSU08180-MON; -.
DR BRENDA; 3.2.1.122; 658.
DR SABIO-RK; P54716; -.
DR EvolutionaryTrace; P54716; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0050081; F:maltose-6'-phosphate glucosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016616; F:oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor; IEA:InterPro.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 3.90.110.10; -; 1.
DR InterPro; IPR019802; GlycHydrolase_4_CS.
DR InterPro; IPR001088; Glyco_hydro_4.
DR InterPro; IPR022616; Glyco_hydro_4_C.
DR InterPro; IPR015955; Lactate_DH/Glyco_Ohase_4_C.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR PANTHER; PTHR32092; PTHR32092; 1.
DR Pfam; PF02056; Glyco_hydro_4; 1.
DR Pfam; PF11975; Glyco_hydro_4C; 1.
DR PRINTS; PR00732; GLHYDRLASE4.
DR SUPFAM; SSF51735; SSF51735; 1.
DR SUPFAM; SSF56327; SSF56327; 1.
DR PROSITE; PS01324; GLYCOSYL_HYDROL_F4; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cobalt; Direct protein sequencing;
KW Glycosidase; Hydrolase; Iron; Manganese; Metal-binding; NAD; Nickel;
KW Reference proteome.
FT CHAIN 1..449
FT /note="Maltose-6'-phosphate glucosidase"
FT /id="PRO_0000169859"
FT ACT_SITE 172
FT /note="Proton donor"
FT ACT_SITE 265
FT /note="Proton acceptor"
FT BINDING 6..72
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT BINDING 95
FT /ligand="substrate"
FT BINDING 149
FT /ligand="substrate"
FT BINDING 171
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT BINDING 202
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT BINDING 285
FT /ligand="substrate"
FT SITE 111
FT /note="Increases basicity of active site Tyr"
FT MUTAGEN 41
FT /note="D->E,G: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9765262"
FT MUTAGEN 111
FT /note="E->D,G: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9765262"
FT MUTAGEN 359
FT /note="E->D,G: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9765262"
FT STRAND 6..11
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 15..17
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 18..27
FT /evidence="ECO:0007829|PDB:1U8X"
FT TURN 28..31
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 34..40
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 44..61
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 65..71
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 73..77
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 81..85
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 91..103
FT /evidence="ECO:0007829|PDB:1U8X"
FT TURN 104..106
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 111..113
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 114..138
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 143..146
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 151..161
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 167..169
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 173..185
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 190..192
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 193..200
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 203..211
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 218..228
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 246..255
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 260..262
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 264..266
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 267..270
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 272..276
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 281..283
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 285..292
FT /evidence="ECO:0007829|PDB:1U8X"
FT TURN 293..300
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 301..307
FT /evidence="ECO:0007829|PDB:1U8X"
FT TURN 319..321
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 322..333
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 337..344
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 356..365
FT /evidence="ECO:0007829|PDB:1U8X"
FT STRAND 368..371
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 379..401
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 404..413
FT /evidence="ECO:0007829|PDB:1U8X"
FT HELIX 420..433
FT /evidence="ECO:0007829|PDB:1U8X"
FT TURN 434..437
FT /evidence="ECO:0007829|PDB:1U8X"
SQ SEQUENCE 449 AA; 50514 MW; A903F7E41CFEF7AB CRC64;
MKKKSFSIVI AGGGSTFTPG IVLMLLDHLE EFPIRKLKLY DNDKERQDRI AGACDVFIRE
KAPDIEFAAT TDPEEAFTDV DFVMAHIRVG KYAMRALDEQ IPLKYGVVGQ ETCGPGGIAY
GMRSIGGVLE ILDYMEKYSP DAWMLNYSNP AAIVAEATRR LRPNSKILNI CDMPVGIEDR
MAQILGLSSR KEMKVRYYGL NHFGWWTSIQ DQEGNDLMPK LKEHVSQYGY IPKTEAEAVE
ASWNDTFAKA RDVQAADPDT LPNTYLQYYL FPDDMVKKSN PNHTRANEVM EGREAFIFSQ
CDMITREQSS ENSEIKIDDH ASYIVDLARA IAYNTGERML LIVENNGAIA NFDPTAMVEV
PCIVGSNGPE PITVGTIPQF QKGLMEQQVS VEKLTVEAWA EKSFQKLWQA LILSKTVPNA
RVARLILEDL VEANKDFWPE LDQSPTRIS
