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 28-JUN-2023, entry version 36. DE RecName: Full=Beta-galactosidase; DE Short=Beta-gal {ECO:0000250|UniProtKB:P19668}; DE EC=3.2.1.23; OS Arthrobacter sp. OC Bacteria; Actinomycetota; Actinomycetes; Micrococcales; Micrococcaceae; OC 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}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; FJ609657; ACU00913.1; -; Genomic_DNA. DR AlphaFoldDB; C7ASJ5; -. DR SMR; C7ASJ5; -. DR CAZy; GH42; Glycoside Hydrolase Family 42. 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 CDD; cd03143; A4_beta-galactosidase_middle_domain; 1. DR Gene3D; 3.40.50.880; -; 1. DR Gene3D; 3.20.20.80; Glycosidases; 1. DR Gene3D; 2.60.40.1180; Golgi alpha-mannosidase II; 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; BETA-GALACTOSIDASE GANA; 1. DR PANTHER; PTHR36447:SF1; BETA-GALACTOSIDASE GANA; 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; (Trans)glycosidases; 1. DR SUPFAM; SSF52317; Class I glutamine amidotransferase-like; 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 //