GMAS_METMY
ID GMAS_METMY Reviewed; 444 AA.
AC A9ZPH9;
DT 04-FEB-2015, integrated into UniProtKB/Swiss-Prot.
DT 26-FEB-2008, sequence version 1.
DT 03-AUG-2022, entry version 41.
DE RecName: Full=Glutamate--methylamine ligase {ECO:0000305};
DE EC=6.3.4.12 {ECO:0000269|PubMed:17284842, ECO:0000269|PubMed:18175924};
DE AltName: Full=Gamma-glutamylmethylamide synthetase {ECO:0000303|PubMed:17284842, ECO:0000303|PubMed:18175924};
DE Short=GMAS {ECO:0000303|PubMed:17284842, ECO:0000303|PubMed:18175924};
DE AltName: Full=Glutamate--ethylamine ligase {ECO:0000305};
DE EC=6.3.1.6 {ECO:0000269|PubMed:17284842, ECO:0000269|PubMed:18175924};
DE AltName: Full=N(5)-ethyl-L-glutamine synthetase {ECO:0000305};
DE AltName: Full=Theanine synthetase {ECO:0000305};
OS Methylovorus mays.
OC Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales;
OC Methylophilaceae; Methylovorus.
OX NCBI_TaxID=184077;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 194-200; 222-241 AND
RP 297-316, FUNCTION, CATALYTIC ACTIVITY, AND BIOTECHNOLOGY.
RC STRAIN=No. 9;
RX PubMed=18175924; DOI=10.1271/bbb.70462;
RA Yamamoto S., Wakayama M., Tachiki T.;
RT "Cloning and expression of Methylovorus mays No. 9 gene encoding gamma-
RT glutamylmethylamide synthetase: an enzyme usable in theanine formation by
RT coupling with the alcoholic fermentation system of baker's yeast.";
RL Biosci. Biotechnol. Biochem. 72:101-109(2008).
RN [2]
RP PROTEIN SEQUENCE OF 1-20, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY
RP REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, INDUCTION, AND BIOTECHNOLOGY.
RC STRAIN=No. 9;
RX PubMed=17284842; DOI=10.1271/bbb.60590;
RA Yamamoto S., Wakayama M., Tachiki T.;
RT "Characterization of theanine-forming enzyme from Methylovorus mays no. 9
RT in respect to utilization of theanine production.";
RL Biosci. Biotechnol. Biochem. 71:545-552(2007).
CC -!- FUNCTION: Catalyzes the formation of N(5)-methyl-L-glutamine from
CC glutamate and methylamine. In vitro, can also use ethylamine,
CC hydroxylamine and ammonia, with 75%, 40% and 1% activity compared to
CC methylamine, respectively. {ECO:0000269|PubMed:17284842,
CC ECO:0000269|PubMed:18175924}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-glutamate + methylamine = ADP + H(+) + N(5)-methyl-L-
CC glutamine + phosphate; Xref=Rhea:RHEA:17117, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29985, ChEBI:CHEBI:30616, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:58200, ChEBI:CHEBI:59338, ChEBI:CHEBI:456216;
CC EC=6.3.4.12; Evidence={ECO:0000269|PubMed:17284842,
CC ECO:0000269|PubMed:18175924};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + ethylamine + L-glutamate = ADP + H(+) + N(5)-ethyl-L-
CC glutamine + phosphate; Xref=Rhea:RHEA:20525, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29985, ChEBI:CHEBI:30616, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:58128, ChEBI:CHEBI:456216, ChEBI:CHEBI:566789;
CC EC=6.3.1.6; Evidence={ECO:0000269|PubMed:17284842,
CC ECO:0000269|PubMed:18175924};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:17284842};
CC -!- ACTIVITY REGULATION: Formation of theanine is repressed by a high
CC concentration of glutamic acid. {ECO:0000269|PubMed:17284842}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.18 mM for methylamine {ECO:0000269|PubMed:17284842};
CC KM=0.57 mM for ethylamine {ECO:0000269|PubMed:17284842};
CC KM=3.9 mM for hydroxylamine {ECO:0000269|PubMed:17284842};
CC KM=84 mM for ammonia {ECO:0000269|PubMed:17284842};
CC KM=1.4 mM for glutamic acid (with methylamine as substrate)
CC {ECO:0000269|PubMed:17284842};
CC KM=1.3 mM for glutamic acid (with ethylamine as substrate)
CC {ECO:0000269|PubMed:17284842};
CC KM=1.3 mM for ATP (with methylamine as substrate)
CC {ECO:0000269|PubMed:17284842};
CC KM=0.63 mM for ATP (with ethylamine as substrate)
CC {ECO:0000269|PubMed:17284842};
CC -!- INDUCTION: By methylamine. {ECO:0000269|PubMed:17284842}.
CC -!- BIOTECHNOLOGY: Could be used for theanine (N(5)-ethyl-L-glutamine)
CC production, by coupling the reaction with the ATP-regeneration system
CC of yeast sugar fermentation. Theanine occurs naturally in the tea plant
CC (Camellia sinensis) and provides flavor to green tea. It has also
CC several favorable physiological effects on mammals.
CC {ECO:0000305|PubMed:17284842, ECO:0000305|PubMed:18175924}.
CC -!- SIMILARITY: Belongs to the glutamine synthetase family. Type 3
CC subfamily. {ECO:0000305}.
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DR EMBL; AB333782; BAF99006.1; -; Genomic_DNA.
DR AlphaFoldDB; A9ZPH9; -.
DR SMR; A9ZPH9; -.
DR BioCyc; MetaCyc:MON-17029; -.
DR BRENDA; 6.3.4.12; 10768.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0004356; F:glutamate-ammonia ligase activity; IEA:InterPro.
DR GO; GO:0047942; F:glutamate-ethylamine ligase activity; IEA:UniProtKB-EC.
DR GO; GO:0047943; F:glutamate-methylamine ligase activity; IEA:UniProtKB-EC.
DR GO; GO:0006542; P:glutamine biosynthetic process; IEA:InterPro.
DR Gene3D; 3.10.20.70; -; 1.
DR InterPro; IPR036651; Gln_synt_N.
DR InterPro; IPR014746; Gln_synth/guanido_kin_cat_dom.
DR InterPro; IPR008146; Gln_synth_cat_dom.
DR InterPro; IPR027303; Gln_synth_gly_rich_site.
DR InterPro; IPR017536; Glutamine_synthetase_typeIII.
DR Pfam; PF00120; Gln-synt_C; 1.
DR SMART; SM01230; Gln-synt_C; 1.
DR SUPFAM; SSF54368; SSF54368; 1.
DR SUPFAM; SSF55931; SSF55931; 1.
DR TIGRFAMs; TIGR03105; gln_synth_III; 1.
DR PROSITE; PS00181; GLNA_ATP; 1.
DR PROSITE; PS51986; GS_BETA_GRASP; 1.
DR PROSITE; PS51987; GS_CATALYTIC; 1.
PE 1: Evidence at protein level;
KW ATP-binding; Direct protein sequencing; Ligase; Magnesium;
KW Nucleotide-binding.
FT CHAIN 1..444
FT /note="Glutamate--methylamine ligase"
FT /id="PRO_0000431889"
FT DOMAIN 14..97
FT /note="GS beta-grasp"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01330"
FT DOMAIN 103..444
FT /note="GS catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01331"
FT CONFLICT 2
FT /note="K -> T (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 14
FT /note="H -> D (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 17
FT /note="K -> V (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 225..226
FT /note="IC -> R (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 236
FT /note="R -> N (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 239
FT /note="N -> G (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 312
FT /note="A -> P (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 314
FT /note="I -> Y (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 444 AA; 48925 MW; F976581C78E7FCB6 CRC64;
MKSLEEAQKF LEDHHVKYVL AQFVDIHGVA KVKSVPASHL NDILTTGAGF AGGAIWGTGI
APNGPDYMAI GELSTLSLIP WQPGYARLVC DGHVNGKPYE FDTRVVLKQQ IARLAEKGWT
LYTGLEPEFS LLKKDEHGAV HPFDDSDTLQ KPCYDYKGIT RHSPFLEKLT ESLVEVGLDI
YQIDHEDANG QFEINYTYAD CLKSADDYIM FKMAASEIAN ELGIICSFMP KPFSNRPGNG
MHMHMSIGDG KKSLFQDDSD PSGLGLSKLA YHFLGGILAH APALAAVCAP TVNSYKRLVV
GRSLSGATWA PAYIAYGNNN RSTLVRIPYG RLELRLPDGS CNPYLATAAV IAAGLDGVAR
ELDPGTGRDD NLYDYSLEQL AEFGIGILPQ NLGEALDALE ADQVIMDAMG PGLSKEFVEL
KRMEWVDYMR HVSDWEINRY VQFY
