GLYOX_BACLI
ID GLYOX_BACLI Reviewed; 369 AA.
AC S5FMM4;
DT 07-JUN-2017, integrated into UniProtKB/Swiss-Prot.
DT 16-OCT-2013, sequence version 1.
DT 03-AUG-2022, entry version 39.
DE RecName: Full=Glycine oxidase {ECO:0000303|PubMed:26920475};
DE Short=GO {ECO:0000303|PubMed:26920475};
DE EC=1.4.3.19 {ECO:0000269|PubMed:26920475};
DE AltName: Full=BliGO {ECO:0000303|PubMed:26920475};
GN Name=thiO {ECO:0000250|UniProtKB:O31616};
OS Bacillus licheniformis.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=1402;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, 3D-STRUCTURE MODELING, PROTEIN
RP ENGINEERING, BIOTECHNOLOGY, AND MUTAGENESIS OF GLY-51.
RC STRAIN=J33-8;
RX PubMed=26920475; DOI=10.1016/j.enzmictec.2015.12.012;
RA Zhang K., Guo Y., Yao P., Lin Y., Kumar A., Liu Z., Wu G., Zhang L.;
RT "Characterization and directed evolution of BliGO, a novel glycine oxidase
RT from Bacillus licheniformis.";
RL Enzyme Microb. Technol. 85:12-18(2016).
CC -!- FUNCTION: Catalyzes the FAD-dependent oxidative deamination of glycine,
CC leading to glyoxylate, ammonia and hydrogen peroxide (PubMed:26920475).
CC Is also able to act on various amines and D-amino acids to yield the
CC corresponding alpha-keto acids, ammonia/amine, and hydrogen peroxide
CC (By similarity). Can also oxidize the herbicide glyphosate (N-
CC phosphonomethylglycine), and thus may be involved in the degradation
CC pathway that allows B.licheniformis J33-8 to grow with glyphosate as
CC the sole source of carbon (PubMed:26920475). Is essential for thiamine
CC biosynthesis since the oxidation of glycine catalyzed by ThiO generates
CC the glycine imine intermediate (dehydroglycine) required for the
CC biosynthesis of the thiazole ring of thiamine pyrophosphate (By
CC similarity). {ECO:0000250|UniProtKB:O31616,
CC ECO:0000269|PubMed:26920475}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=glycine + H2O + O2 = glyoxylate + H2O2 + NH4(+);
CC Xref=Rhea:RHEA:11532, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16240, ChEBI:CHEBI:28938, ChEBI:CHEBI:36655,
CC ChEBI:CHEBI:57305; EC=1.4.3.19;
CC Evidence={ECO:0000269|PubMed:26920475};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=glyphosate + H2O + O2 = aminomethylphosphonate + glyoxylate +
CC H(+) + H2O2; Xref=Rhea:RHEA:52740, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:36655, ChEBI:CHEBI:133673, ChEBI:CHEBI:133674;
CC Evidence={ECO:0000269|PubMed:26920475};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-ethylglycine + O2 = ethylamine + glyoxylate + H2O2;
CC Xref=Rhea:RHEA:12472, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16240, ChEBI:CHEBI:36655, ChEBI:CHEBI:57440,
CC ChEBI:CHEBI:566789; EC=1.4.3.19;
CC Evidence={ECO:0000250|UniProtKB:O31616};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + O2 + sarcosine = glyoxylate + H2O2 + methylamine;
CC Xref=Rhea:RHEA:15165, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16240, ChEBI:CHEBI:36655, ChEBI:CHEBI:57433,
CC ChEBI:CHEBI:59338; EC=1.4.3.19;
CC Evidence={ECO:0000250|UniProtKB:O31616};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-alanine + H2O + O2 = H2O2 + NH4(+) + pyruvate;
CC Xref=Rhea:RHEA:22688, ChEBI:CHEBI:15361, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16240, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:57416; EC=1.4.3.19;
CC Evidence={ECO:0000250|UniProtKB:O31616};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000250|UniProtKB:O31616, ECO:0000305|PubMed:26920475};
CC Note=Binds 1 FAD per subunit. {ECO:0000250|UniProtKB:O31616};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.9 mM for glycine {ECO:0000269|PubMed:26920475};
CC KM=11.22 mM for glyphosate {ECO:0000269|PubMed:26920475};
CC Note=kcat is 0.31 sec(-1) with glycine as substrate. kcat is 0.08
CC sec(-1) with glyphosate as substrate. {ECO:0000269|PubMed:26920475};
CC pH dependence:
CC Optimum pH is 8.5. Displays over 60% of maximum activity at pH 8.0-
CC 10. {ECO:0000269|PubMed:26920475};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius. Retains 60% of the maximum
CC activity at 0 degree Celsius, suggesting it is a cold-adapted enzyme.
CC {ECO:0000269|PubMed:26920475};
CC -!- PATHWAY: Cofactor biosynthesis; thiamine diphosphate biosynthesis.
CC {ECO:0000250|UniProtKB:O31616}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000250|UniProtKB:O31616}.
CC -!- BIOTECHNOLOGY: This glyphosate-degrading enzyme could be of potential
CC use in agricultural biotechnology for the development of new-generation
CC herbicide-resistant plants. The SCF-4 mutant obtained by engineering
CC shows significant improved catalytic efficiency towards glyphosate, and
CC a better affinity to glyphosate than that reported for glyphosate
CC oxidoreductase GOX in a Monsanto's patent.
CC {ECO:0000305|PubMed:26920475}.
CC -!- SIMILARITY: Belongs to the DAO family. ThiO subfamily. {ECO:0000305}.
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DR EMBL; KC831746; AGQ46835.1; -; Genomic_DNA.
DR RefSeq; WP_003180677.1; NZ_UAQA01000001.1.
DR AlphaFoldDB; S5FMM4; -.
DR SMR; S5FMM4; -.
DR GeneID; 66216601; -.
DR PATRIC; fig|1402.62.peg.4127; -.
DR OMA; GCGTLWL; -.
DR BRENDA; 1.4.3.19; 669.
DR UniPathway; UPA00060; -.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro.
DR GO; GO:0043799; F:glycine oxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0009228; P:thiamine biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0009229; P:thiamine diphosphate biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR006076; FAD-dep_OxRdtase.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR012727; Gly_oxidase_ThiO.
DR Pfam; PF01266; DAO; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR TIGRFAMs; TIGR02352; thiamin_ThiO; 1.
PE 1: Evidence at protein level;
KW FAD; Flavoprotein; Oxidoreductase; Thiamine biosynthesis.
FT CHAIN 1..369
FT /note="Glycine oxidase"
FT /id="PRO_0000440179"
FT BINDING 14..15
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 34..35
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 42..43
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 47..49
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 174
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 302
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 327..333
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT BINDING 329
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:O31616"
FT MUTAGEN 51
FT /note="G->S: Shows 4.3- and 107-fold increase of affinity
FT to glyphosate and glycine, respectively. Shows 7.1- and 8-
FT fold increase of affinity and catalytic efficiency to
FT glyphosate, respectively, while the substrate affinity to
FT glycine decreases 235-fold and catalytic efficiency
FT decreases 113-fold; when associated with R-54, R-81, C-202,
FT V-332 and V-342."
FT /evidence="ECO:0000269|PubMed:26920475"
FT MUTAGEN 54
FT /note="A->R: Shows 7.1- and 8-fold increase of affinity and
FT catalytic efficiency to glyphosate, respectively, while the
FT substrate affinity to glycine decreases 235-fold and
FT catalytic efficiency decreases 113-fold; when associated
FT with S-51, R-81, C-202, V-332 and V-342."
FT /evidence="ECO:0000269|PubMed:26920475"
FT MUTAGEN 81
FT /note="K->R: Shows 7.1- and 8-fold increase of affinity and
FT catalytic efficiency to glyphosate, respectively, while the
FT substrate affinity to glycine decreases 235-fold and
FT catalytic efficiency decreases 113-fold; when associated
FT with S-51, R-54, C-202, V-332 and V-342."
FT /evidence="ECO:0000269|PubMed:26920475"
FT MUTAGEN 202
FT /note="S->C: Shows 7.1- and 8-fold increase of affinity and
FT catalytic efficiency to glyphosate, respectively, while the
FT substrate affinity to glycine decreases 235-fold and
FT catalytic efficiency decreases 113-fold; when associated
FT with S-51, R-54, R-81, V-332 and V-342."
FT /evidence="ECO:0000269|PubMed:26920475"
FT MUTAGEN 332
FT /note="I->V: Shows 7.1- and 8-fold increase of affinity and
FT catalytic efficiency to glyphosate, respectively, while the
FT substrate affinity to glycine decreases 235-fold and
FT catalytic efficiency decreases 113-fold; when associated
FT with S-51, R-54, R-81, C-202 and V-342."
FT /evidence="ECO:0000269|PubMed:26920475"
FT MUTAGEN 342
FT /note="M->V: Shows 7.1- and 8-fold increase of affinity and
FT catalytic efficiency to glyphosate, respectively, while the
FT substrate affinity to glycine decreases 235-fold and
FT catalytic efficiency decreases 113-fold; when associated
FT with S-51, R-54, R-81, C-202 and V-332."
FT /evidence="ECO:0000269|PubMed:26920475"
SQ SEQUENCE 369 AA; 40168 MW; 07737643A01AB54C CRC64;
MRKRYDTIVI GGGIIGTSIA YHLAKAGKKT AVFESGEVGK KATSAAAGML GAHAECDKPG
TFFEFARASQ KAYKRLTGEL KDISGIDIRR HDGGILKLAF SESDREHLMQ MGALDSVEWL
EADEVYKLEP NAGKGILGAN FIRDDVHVEP AAVCRAFARG ARMLGADVFE YTPVLSIESE
AGAVRVTSAS GTAEAEHAVI ASGVWSGALF KQIGLDKRFY PVKGECLSVW NDGISLTRTL
YHDHCYIVPR HSGRLVVGAT MKPGDWNEQP ELGGIEELIR KAKSMLPGIE SMKIDQCWAG
LRPETGDGNP YIGRHPENDR ILFAAGHFRN GILLAPATGE MMADMILGNP VKTEWIEAFK
AERKEAVHR
