BGAL2_BACLD
ID BGAL2_BACLD Reviewed; 673 AA.
AC Q65CX4; Q62NE9;
DT 24-MAR-2009, integrated into UniProtKB/Swiss-Prot.
DT 24-MAR-2009, sequence version 2.
DT 03-AUG-2022, entry version 102.
DE RecName: Full=Beta-galactosidase GalA;
DE Short=Beta-gal;
DE EC=3.2.1.23;
DE AltName: Full=Beta-1,4-galactooligomerase;
DE AltName: Full=Galactooligomerase;
GN Name=lacA {ECO:0000303|PubMed:20852995}; Synonyms=galO, ganA;
GN OrderedLocusNames=BLi04277, BL00264;
OS Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / CCUG 7422 /
OS NBRC 12200 / NCIMB 9375 / NCTC 10341 / NRRL NRS-1264 / Gibson 46).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=279010;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 14580 / DSM 13 / JCM 2505 / CCUG 7422 / NBRC 12200 / NCIMB 9375
RC / NCTC 10341 / NRRL NRS-1264 / Gibson 46;
RX PubMed=15383718; DOI=10.1159/000079829;
RA Veith B., Herzberg C., Steckel S., Feesche J., Maurer K.H., Ehrenreich P.,
RA Baeumer S., Henne A., Liesegang H., Merkl R., Ehrenreich A., Gottschalk G.;
RT "The complete genome sequence of Bacillus licheniformis DSM13, an organism
RT with great industrial potential.";
RL J. Mol. Microbiol. Biotechnol. 7:204-211(2004).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 14580 / DSM 13 / JCM 2505 / CCUG 7422 / NBRC 12200 / NCIMB 9375
RC / NCTC 10341 / NRRL NRS-1264 / Gibson 46;
RX PubMed=15461803; DOI=10.1186/gb-2004-5-10-r77;
RA Rey M.W., Ramaiya P., Nelson B.A., Brody-Karpin S.D., Zaretsky E.J.,
RA Tang M., Lopez de Leon A., Xiang H., Gusti V., Clausen I.G., Olsen P.B.,
RA Rasmussen M.D., Andersen J.T., Joergensen P.L., Larsen T.S., Sorokin A.,
RA Bolotin A., Lapidus A., Galleron N., Ehrlich S.D., Berka R.M.;
RT "Complete genome sequence of the industrial bacterium Bacillus
RT licheniformis and comparisons with closely related Bacillus species.";
RL Genome Biol. 5:R77.1-R77.12(2004).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, SUBUNIT, AND BIOTECHNOLOGY.
RX PubMed=20852995; DOI=10.1007/s00253-010-2862-2;
RA Juajun O., Nguyen T.H., Maischberger T., Iqbal S., Haltrich D.,
RA Yamabhai M.;
RT "Cloning, purification, and characterization of beta-galactosidase from
RT Bacillus licheniformis DSM 13.";
RL Appl. Microbiol. Biotechnol. 89:645-654(2011).
CC -!- FUNCTION: Catalyzes the hydrolysis of lactose to its constituent
CC monosaccharides glucose and galactose. Possesses a low level of
CC transgalactosylation activity for the production of galacto-
CC oligosaccharides (GOS) from lactose. {ECO:0000269|PubMed:20852995}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal non-reducing beta-D-galactose residues
CC in beta-D-galactosides.; EC=3.2.1.23;
CC Evidence={ECO:0000269|PubMed:20852995};
CC -!- ACTIVITY REGULATION: Inhibited by hydrolysis end products D-galactose
CC and D-glucose. The hydrolysis of o-nitrophenyl-beta-D-galactopyranoside
CC (ONPG) is slightly activated by monovalent ions, Na(+) and K(+).
CC Concentrations of these ions in the range of 1-100 mM exert the
CC stimulating effects. The presence of 1 mM Mn(2+) together with the
CC presence of 10 mM Na(+) slightly stimulates the activity, while
CC presence of 10 mM Mn(2+) inhibits the activity by about 40%.
CC {ECO:0000269|PubMed:20852995}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=13.7 mM for ONPG (at 30 degrees Celsius and pH 6.5)
CC {ECO:0000269|PubMed:20852995};
CC KM=169 mM for lactose (at 30 degrees Celsius and pH 6.5)
CC {ECO:0000269|PubMed:20852995};
CC Vmax=299 umol/min/mg enzyme with ONPG as substrate (at 30 degrees
CC Celsius and pH 6.5) {ECO:0000269|PubMed:20852995};
CC Vmax=13 umol/min/mg enzyme with lactose as substrate (at 30 degrees
CC Celsius and pH 6.5) {ECO:0000269|PubMed:20852995};
CC pH dependence:
CC Optimum pH is 6.5 for both lactose and ONPG hydrolysis. Stable at pH
CC 5-8 and most stable at 6.5, retaining more than 90% and 80% of its
CC activity when incubated at pH 6.5 and 37 degrees Celsius for 5 days
CC and 1 month, respectively. {ECO:0000269|PubMed:20852995};
CC Temperature dependence:
CC Optimum temperature of the activity is 50 degrees Celsius when using
CC both lactose and ONPG as substrates under 10 minutes assay
CC conditions. Stable over a wide range of temperatures (4-42 degrees
CC Celsius), and when kept at these temperatures up to 1 month. Most
CC stable at 37 degrees Celsius, retaining 90% of its activity after 1
CC month at this temperature. Half-life time of activity of
CC approximately 7 days, 5 hours, and 30 minutes at 55, 60 and 65
CC degrees Celsius, respectively. Approximately 45% of lactose is
CC hydrolyzed within the first 3 hours of the reaction at 60 degrees
CC Celsius, while about 20% is cleaved at 37 degrees Celsius when
CC employing initial lactose concentration of 50 g/l at pH 6.5. For
CC initial lactose concentrations of 200 and 50 g/l and at temperature
CC of 60 degrees Celsius, the maximum GOS yields are approximately 12%
CC and 7%, respectively. At the initial lactose concentration of 200
CC g/l, the GOS yields obtained at 60 degrees Celsius are significantly
CC higher than at 37 degrees Celsius, with approximately 12% and 5% of
CC total sugars, respectively. {ECO:0000269|PubMed:20852995};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:20852995}.
CC -!- BIOTECHNOLOGY: Has potential for partial lactose removal in food
CC products and improving the quality of dairy products by increasing
CC their solubility and sweetness. {ECO:0000269|PubMed:20852995}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 42 family. {ECO:0000305}.
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DR EMBL; CP000002; AAU25712.1; -; Genomic_DNA.
DR EMBL; AE017333; AAU43090.1; -; Genomic_DNA.
DR AlphaFoldDB; Q65CX4; -.
DR SMR; Q65CX4; -.
DR STRING; 279010.BL00264; -.
DR CAZy; GH42; Glycoside Hydrolase Family 42.
DR EnsemblBacteria; AAU25712; AAU25712; BL00264.
DR KEGG; bld:BLi04277; -.
DR KEGG; bli:BL00264; -.
DR eggNOG; COG1874; Bacteria.
DR HOGENOM; CLU_012430_1_1_9; -.
DR Proteomes; UP000000606; Chromosome.
DR GO; GO:0009341; C:beta-galactosidase complex; IEA:InterPro.
DR GO; GO:0004565; F:beta-galactosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0006012; P:galactose metabolic process; IEA:InterPro.
DR Gene3D; 2.60.40.1180; -; 1.
DR Gene3D; 3.40.50.880; -; 1.
DR InterPro; IPR013739; Beta_galactosidase_C.
DR InterPro; IPR013738; Beta_galactosidase_Trimer.
DR InterPro; IPR029062; Class_I_gatase-like.
DR InterPro; IPR003476; Glyco_hydro_42.
DR InterPro; IPR013529; Glyco_hydro_42_N.
DR InterPro; IPR013780; Glyco_hydro_b.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR PANTHER; PTHR36447; PTHR36447; 1.
DR Pfam; PF02449; Glyco_hydro_42; 1.
DR Pfam; PF08533; Glyco_hydro_42C; 1.
DR Pfam; PF08532; Glyco_hydro_42M; 1.
DR PIRSF; PIRSF001084; B-galactosidase; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR SUPFAM; SSF52317; SSF52317; 1.
PE 1: Evidence at protein level;
KW Glycosidase; Hydrolase; Metal-binding; Reference proteome; Zinc.
FT CHAIN 1..673
FT /note="Beta-galactosidase GalA"
FT /id="PRO_0000367027"
FT ACT_SITE 144
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT ACT_SITE 308
FT /note="Nucleophile"
FT /evidence="ECO:0000250"
FT BINDING 105
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 109
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250"
FT BINDING 143
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 149
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250"
FT BINDING 151
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250"
FT BINDING 154
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250"
FT BINDING 316
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 356..359
FT /ligand="substrate"
FT /evidence="ECO:0000250"
SQ SEQUENCE 673 AA; 77497 MW; D21163244C361A34 CRC64;
MLHGGDYNPD QWLDRPDILA DDIKLMKLAH TNTFSVGIFS WSALEPEEGV YTFEWLDDIF
ESIHRNGGRI ILATPSGARP AWLSQKYPEV LRVNAERVKQ LHGGRHNHCF TSYVYREKTK
EINRMLAERY GSQHALLMWH VSNEYGGECH CDQCQHAFRD WLKKKYNHDI KSLNDAWWTP
FWSHTFNDWS QIESPSPIGE NAVHGLNLDW RRFVTDQTIS FFQNEIVPLK EITPNIPITT
NFMADTHDLI PFQGLDYSKF AKHLDVISWD AYPAWHNDWE STADLAMKVG FINDLYRSLK
QQPFLLMEST PSAVNWHDFN KAKRPGMHLL SSVQMIAHGS DSILYFQWRK SRGSSEKFHG
AVVGHDNCSE NRVFKEVAKV GQTLEALSEV TGTIRPADVA ILYDWENHWA LQDAQGFGMK
TKRYPQTLHE HYRAFWERDI PVDVITKEQD FSSYRLLIVP MLYLASEETI ARLKAFAANG
GTLVMTYISG IVNESDLTYL GGWPKDLQEM FGMEPVETDT LYPGDKNAVR YQNRSYELKD
YATVLKLSTA DPEGFYEDDF YADTTAVTSH PYKQGKTYYI GARLSSQFHR DFYGTLIKEL
AIQPALDVKH QPGVSVQVRQ DEENDYIFIM NFTEKRQPVV LASAVKDMLT GETLAGEVTL
EKYEARIAVK AKE
