BGAL2_ARTSP
ID BGAL2_ARTSP Reviewed; 694 AA.
AC C7ASJ5;
DT 03-MAY-2011, integrated into UniProtKB/Swiss-Prot.
DT 22-SEP-2009, sequence version 1.
DT 03-AUG-2022, entry version 31.
DE RecName: Full=Beta-galactosidase;
DE Short=Beta-gal {ECO:0000250|UniProtKB:P19668};
DE EC=3.2.1.23;
OS Arthrobacter sp.
OC Bacteria; Actinobacteria; Micrococcales; Micrococcaceae; Arthrobacter.
OX NCBI_TaxID=1667;
RN [1] {ECO:0000305, ECO:0000312|EMBL:ACU00913.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND BIOTECHNOLOGY.
RC STRAIN=32c {ECO:0000312|EMBL:ACU00913.1};
RX PubMed=19631003; DOI=10.1186/1471-2180-9-151;
RA Hildebrandt P., Wanarska M., Kur J.;
RT "A new cold-adapted beta-D-galactosidase from the Antarctic Arthrobacter
RT sp. 32c - gene cloning, overexpression, purification and properties.";
RL BMC Microbiol. 9:151-151(2009).
CC -!- FUNCTION: Hydrolyzes p-nitrophenyl-beta-D-galactopyranoside (PNPG), o-
CC nitrophenyl-beta-D-galactopyranoside (ONPG) and chromogen 5-bromo-4-
CC chloro-3-indolyl-beta-D-galactopyranoside (X-gal), with highest
CC activity against PNPG. Also acts on p-nitrophenyl-beta-D-
CC glucopyranoside (PNPGlu) and o-nitrophenyl-beta-D-glucopyranoside
CC (ONPGlu), but with significantly lower activity.
CC {ECO:0000269|PubMed:19631003}.
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:19631003};
CC -!- ACTIVITY REGULATION: Strongly inhibited by glucose. No activity is lost
CC during treatment with 100 mM EDTA after 2 hours. Activity not
CC considerably affected by metal ions (5 mM), including Na(+), K(+),
CC Mg(2+), Co(2+) and Ca(2+). Completely inhibited by Cu(2+) and Zn(2+) (5
CC mM) and is strongly inhibited by Mn(2+) (11%), Fe(2+) (25%) and Ni(2+)
CC (38%) in comparison with the activity in the absence of cations (100%).
CC Activity not affected by dithiothreitol, beta-mercaptoethanol and L-
CC cysteine whereas reduced glutathione almost completely inactivates it.
CC With ONPG as substrate, the addition of ethanol up to 20% still
CC slightly stimulates activity. The activity increases up to 120% in the
CC presence of 8% v/v ethanol at pH 5.5. {ECO:0000269|PubMed:19631003}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.75 mM for ONPG (at 10 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=4.86 mM for ONPG (at 20 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=3.46 mM for ONPG (at 30 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=3.15 mM for ONPG (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=2.62 mM for ONPG (at 50 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=5.11 mM for ONPG (at 55 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=77.54 mM for lactose (at 10 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=67.82 mM for lactose (at 20 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=52.67 mM for lactose (at 30 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=44.31 mM for lactose (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC KM=39.73 mM for lactose (at 50 degrees Celsius)
CC {ECO:0000269|PubMed:19631003};
CC pH dependence:
CC Optimum pH is 6.5 with ONPG as substrate. Over 90% of activity in the
CC pH range 6.5-8.5. Highly efficient at pH range 4.5-9.5.
CC {ECO:0000269|PubMed:19631003};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius with ONPG as substrate.
CC Active at 4-8 degrees Celsius. 60% of the maximum activity is
CC detected at 25 degrees Celsius and 15% at 0 degrees Celsius. Over 50%
CC activity at 30 degrees Celsius. {ECO:0000269|PubMed:19631003};
CC -!- SUBUNIT: Homotrimer. {ECO:0000269|PubMed:19631003}.
CC -!- BIOTECHNOLOGY: Has potential use in lactose removal in milk and dairy
CC products at low temperatures and for cheese whey bioremediation
CC processes with simultaneous bio-ethanol production.
CC {ECO:0000269|PubMed:19631003}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 42 family. {ECO:0000255}.
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DR EMBL; FJ609657; ACU00913.1; -; Genomic_DNA.
DR AlphaFoldDB; C7ASJ5; -.
DR SMR; C7ASJ5; -.
DR CAZy; GH42; Glycoside Hydrolase Family 42.
DR PRIDE; C7ASJ5; -.
DR BRENDA; 3.2.1.23; 457.
DR GO; GO:0009341; C:beta-galactosidase complex; IEA:InterPro.
DR GO; GO:0004565; F:beta-galactosidase activity; IEA:UniProtKB-EC.
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.
FT CHAIN 1..694
FT /note="Beta-galactosidase"
FT /id="PRO_0000407681"
FT REGION 1..31
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 183
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:O69315"
FT ACT_SITE 341
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:O69315"
FT BINDING 144
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:O69315"
FT BINDING 182
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:O69315"
FT BINDING 349
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:O69315"
FT BINDING 389..392
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:O69315"
SQ SEQUENCE 694 AA; 75915 MW; 2A352EDCF797BE74 CRC64;
MGKRFPSGWF SPRVHPPRRQ RSPMTNQATP GTASVWNNIE GIGFGGDYNP EQWPVSVRLE
DLELMQEAGV NFLSVGIFSW ALLEPAEGQY DFGWLDDVMD NLHGIGVKVA LATATAAPPA
WLVRKHPEIL PVTADGTTLG PGSRRHYTPS SAVYRKYAAG ITRVLAERYK DHPALALWHV
DNELGCHVSE FYGEEDAAAF RLWLERRYGT IDALNAAWGT AFWSQHYGSF EEILPPGVAP
STLNPGQQLD FQRFNSWALM DYYRSLVAVL REVTPAVPCT TNLMASSATK SMDYFSWAKD
LDVIANDHYL VAADPERHIE LAFSADLTRG IAGGDPWILM EHSTSAVNWQ PRNQPKMPGE
MLRNSLAHVA RGADAVMFFQ WRQSFAGSEK FHSAMVPHGG RDTRVWREVV DLGAALQLLA
PVRGSRVESR AAIVFDYEAW WASEIDSKPS IDVRYLDLLR AFHRSLFLRG VSVDMVHPSA
SLDGYDLVLV CTLYSVTDEA AANIAAAAAG GATVLVSYFS GITDEKDHVR LGGYPGAFRE
LLGVRVEEFH PLLAGSQLKL SDGTVSSIWS EHVHLDGAEA FQTFTGYPLE GVPSLTRRAV
GTGAAWYLAT FPDRDGIESL VDRLLAESGV SPVAEADAGV ELTRRRSADG GSFLFAINHT
RAAASVRASG TDVLSGERFT GTVEAGSVAV IAED
