MALH_CLOAB
ID MALH_CLOAB Reviewed; 441 AA.
AC Q97LM4;
DT 15-FEB-2005, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2001, sequence version 1.
DT 03-AUG-2022, entry version 107.
DE RecName: Full=Maltose-6'-phosphate glucosidase MalH;
DE EC=3.2.1.122;
DE AltName: Full=6-phospho-alpha-glucosidase;
DE AltName: Full=6-phospho-glucosidase;
DE AltName: Full=Maltose-6-phosphate hydrolase;
GN Name=malH; OrderedLocusNames=CA_C0533;
OS Clostridium acetobutylicum (strain ATCC 824 / DSM 792 / JCM 1419 / LMG 5710
OS / VKM B-1787).
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae;
OC Clostridium.
OX NCBI_TaxID=272562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787;
RX PubMed=11781805; DOI=10.1038/sj/jim/7000125;
RA Tangney M., Winters G.T., Mitchell W.J.;
RT "Characterization of a maltose transport system in Clostridium
RT acetobutylicum ATCC 824.";
RL J. Ind. Microbiol. Biotechnol. 27:298-306(2001).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787;
RX PubMed=11466286; DOI=10.1128/jb.183.16.4823-4838.2001;
RA Noelling J., Breton G., Omelchenko M.V., Makarova K.S., Zeng Q., Gibson R.,
RA Lee H.M., Dubois J., Qiu D., Hitti J., Wolf Y.I., Tatusov R.L., Sabathe F.,
RA Doucette-Stamm L.A., Soucaille P., Daly M.J., Bennett G.N., Koonin E.V.,
RA Smith D.R.;
RT "Genome sequence and comparative analysis of the solvent-producing
RT bacterium Clostridium acetobutylicum.";
RL J. Bacteriol. 183:4823-4838(2001).
RN [3]
RP PROTEIN SEQUENCE OF 1-29, CHARACTERIZATION, KINETIC PARAMETERS, MUTAGENESIS
RP OF CYS-169; ASP-170; MET-171 AND PRO-172, AND MASS SPECTROMETRY.
RC STRAIN=ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787;
RX PubMed=14570887; DOI=10.1074/jbc.m310733200;
RA Thompson J., Hess S., Pikis A.;
RT "Genes malh and pagl of Clostridium acetobutylicum ATCC 824 encode
RT NAD+- and Mn2+-dependent phospho-alpha-glucosidase(s).";
RL J. Biol. Chem. 279:1553-1561(2004).
CC -!- FUNCTION: Catalyzes the hydrolysis of O-alpha-linked disaccharide 6-
CC phosphates, including maltose-6'P and all five phosphorylated isomers
CC of sucrose, but not sucrose-6P. Does not hydrolyze beta-linked
CC disaccharide 6-phosphates such as cellobiose-6'P and gentiobiose-6'P.
CC Is involved in the dissimilation of maltose and related O-alpha-linked
CC glucosides produced via the phosphoenolpyruvate-dependent sugar
CC phosphotransferase system (PEP-PTS).
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=NAD(+); Xref=ChEBI:CHEBI:57540;
CC Note=Binds 1 NAD(+) per subunit.;
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Note=Binds 1 Mn(2+) ion per subunit.;
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=28.4 uM for p-nitrophenyl-alpha-D-glucopyranoside 6-phosphate
CC {ECO:0000269|PubMed:14570887};
CC KM=1.92 mM for trehalulose-6'P {ECO:0000269|PubMed:14570887};
CC KM=0.82 mM for turanose-6'P {ECO:0000269|PubMed:14570887};
CC KM=1.11 mM for maltulose-6'P {ECO:0000269|PubMed:14570887};
CC KM=1.87 mM for leucrose-6'P {ECO:0000269|PubMed:14570887};
CC KM=2.47 mM for palatinose-6'P {ECO:0000269|PubMed:14570887};
CC KM=1.95 mM for maltose-6'P {ECO:0000269|PubMed:14570887};
CC Vmax=5.2 umol/min/mg enzyme with p-nitrophenyl-alpha-D-
CC glucopyranoside 6-phosphate as substrate
CC {ECO:0000269|PubMed:14570887};
CC Vmax=0.43 umol/min/mg enzyme with trehalulose-6'P as substrate
CC {ECO:0000269|PubMed:14570887};
CC Vmax=0.82 umol/min/mg enzyme with turanose-6'P as substrate
CC {ECO:0000269|PubMed:14570887};
CC Vmax=0.65 umol/min/mg enzyme with maltulose-6'P as substrate
CC {ECO:0000269|PubMed:14570887};
CC Vmax=0.12 umol/min/mg enzyme with leucrose-6'P as substrate
CC {ECO:0000269|PubMed:14570887};
CC Vmax=0.60 umol/min/mg enzyme with palatinose-6'P as substrate
CC {ECO:0000269|PubMed:14570887};
CC Vmax=0.49 umol/min/mg enzyme with maltose-6'P as substrate
CC {ECO:0000269|PubMed:14570887};
CC -!- SUBUNIT: Homotetramer.
CC -!- INDUCTION: By maltose and other alpha-glucosides, except sucrose.
CC -!- MASS SPECTROMETRY: Mass=49973; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:14570887};
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 4 family. {ECO:0000305}.
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DR EMBL; AF290982; AAK69556.1; -; Genomic_DNA.
DR EMBL; AE001437; AAK78512.1; -; Genomic_DNA.
DR PIR; E96965; E96965.
DR RefSeq; NP_347172.1; NC_003030.1.
DR RefSeq; WP_010963854.1; NC_003030.1.
DR AlphaFoldDB; Q97LM4; -.
DR SMR; Q97LM4; -.
DR STRING; 272562.CA_C0533; -.
DR CAZy; GH4; Glycoside Hydrolase Family 4.
DR EnsemblBacteria; AAK78512; AAK78512; CA_C0533.
DR GeneID; 44997043; -.
DR KEGG; cac:CA_C0533; -.
DR PATRIC; fig|272562.8.peg.736; -.
DR eggNOG; COG1486; Bacteria.
DR HOGENOM; CLU_045951_2_0_9; -.
DR OMA; KYPMREK; -.
DR OrthoDB; 277080at2; -.
DR SABIO-RK; Q97LM4; -.
DR Proteomes; UP000000814; 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 Carbohydrate metabolism; Direct protein sequencing; Glycosidase; Hydrolase;
KW Manganese; Metal-binding; NAD; Reference proteome.
FT CHAIN 1..441
FT /note="Maltose-6'-phosphate glucosidase MalH"
FT /id="PRO_0000169863"
FT ACT_SITE 170
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT ACT_SITE 264
FT /note="Proton acceptor"
FT /evidence="ECO:0000250"
FT BINDING 4..70
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 93
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 147
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 169
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250"
FT BINDING 200
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250"
FT BINDING 284
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT SITE 109
FT /note="Increases basicity of active site Tyr"
FT /evidence="ECO:0000250"
FT MUTAGEN 169
FT /note="C->S: Loss of activity."
FT /evidence="ECO:0000269|PubMed:14570887"
FT MUTAGEN 170
FT /note="D->N: Loss of activity."
FT /evidence="ECO:0000269|PubMed:14570887"
FT MUTAGEN 171
FT /note="M->V: Highly reduced activity."
FT /evidence="ECO:0000269|PubMed:14570887"
FT MUTAGEN 172
FT /note="P->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:14570887"
SQ SEQUENCE 441 AA; 49972 MW; B525EE15EC0CD69E CRC64;
MKKFSVVIAG GGSTFTPGIV LMLLDNMDKF PIRKLKFYDN DKERQAIVAG ACEIILKEKA
PEIEFLATTN PKEAFTDVDF VMAHIRVGKY AMRELDEKIP LKYGVVGQET CGPGGIAYGM
RSIGGVIEIL DYMEKYSPNA WMLNYSNPAA IVAEATRKLR PNSKILNICD MPIGIETRMA
EILGLESRKE MTVKYYGLNH FGWWSDIRDK DGNDLMPKLK EHVKKYGYVA ENGDTQHTDA
SWNDTFAKAK DVYAVDPSTL PNTYLKYYLF PDYVVEHSNK EYTRANEVMD GREKFVFGEC
KKVIENQSTK GCKMEIDEHA SYIVDLARAI SYNTHERMLL IVPNNGSIEN FDSTGMVEIP
CIVGSNGPEP LTMGKIPQFQ KGLMEQQVSV EKLVVEAWKE KSYQKLWQAI TLSRTVPSAK
VAKQILDELI EVNKDYWPEL N
