CEO2_LACLL
ID CEO2_LACLL Reviewed; 313 AA.
AC P15244;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1997, sequence version 2.
DT 03-AUG-2022, entry version 102.
DE RecName: Full=N(5)-(carboxyethyl)ornithine synthase;
DE Short=CEOS;
DE EC=1.5.1.24;
DE AltName: Full=N(5)-(L-1-carboxyethyl)-L-ornithine:NADP(+) oxidoreductase;
GN Name=ceo;
OS Lactococcus lactis subsp. lactis (Streptococcus lactis).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Lactococcus.
OX NCBI_TaxID=1360;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND MUTAGENESIS OF ARG-15.
RC STRAIN=K1-23; TRANSPOSON=Tn5306;
RX PubMed=7744873; DOI=10.1074/jbc.270.20.12226;
RA Donkersloot J.A., Thompson J.;
RT "Cloning, expression, sequence analysis, and site-directed mutagenesis of
RT the Tn5306-encoded N(5)-(carboxyethyl)ornithine synthase from Lactococcus
RT lactis K1.";
RL J. Biol. Chem. 270:12226-12234(1995).
RN [2]
RP PROTEIN SEQUENCE OF 1-37, FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE
RP SPECIFICITY, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=K1;
RX PubMed=2498334; DOI=10.1016/s0021-9258(18)60572-1;
RA Thompson J.;
RT "N(5)-(L-1-carboxyethyl)-L-ornithine:NADP(+) oxidoreductase from
RT Streptococcus lactis. Purification and partial characterization.";
RL J. Biol. Chem. 264:9592-9601(1989).
RN [3]
RP PROTEIN SEQUENCE OF 256-263, SUBUNIT, AND MASS SPECTROMETRY.
RC STRAIN=K1;
RX PubMed=10548058; DOI=10.1110/ps.8.10.2121;
RA Sackett D.L., Ruvinov S.B., Thompson J.;
RT "N(5)-(L-1-carboxyethyl)-L-ornithine synthase: physical and spectral
RT characterization of the enzyme and its unusual low pKa fluorescent tyrosine
RT residues.";
RL Protein Sci. 8:2121-2129(1999).
RN [4]
RP FOLDING STUDIES.
RC STRAIN=K1;
RX PubMed=10525296; DOI=10.1006/abbi.1999.1429;
RA Ruvinov S.B., Thompson J., Sackett D.L., Ginsburg A.;
RT "Tetrameric N(5)-(L-1-carboxyethyl)-L-ornithine synthase: guanidine. HCl-
RT induced unfolding and a low temperature requirement for refolding.";
RL Arch. Biochem. Biophys. 371:115-123(1999).
CC -!- FUNCTION: Catalyzes the NADPH-dependent reductive condensation between
CC pyruvic acid and the side chain amino group of L-ornithine to form
CC N(5)-(L-1-carboxyethyl)-L-ornithine. To a lesser extent, can also use
CC L-lysine as substrate (yielding N(6)-(L-1-carboxyethyl)-L-lysine). NADH
CC cannot replace NADPH in the condensation reaction.
CC {ECO:0000269|PubMed:2498334}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N(5)-[1(S)-1-carboxyethyl]-L-ornithine + NADP(+) = H(+)
CC + L-ornithine + NADPH + pyruvate; Xref=Rhea:RHEA:18661,
CC ChEBI:CHEBI:15361, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:46911, ChEBI:CHEBI:57783, ChEBI:CHEBI:57889,
CC ChEBI:CHEBI:58349; EC=1.5.1.24;
CC Evidence={ECO:0000269|PubMed:2498334};
CC -!- ACTIVITY REGULATION: Is potently inhibited by the reaction product
CC N(5)-(L-1-carboxyethyl)-L-ornithine. {ECO:0000269|PubMed:2498334}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.3 mM for L-ornithine {ECO:0000269|PubMed:2498334};
CC KM=18.2 mM for L-lysine {ECO:0000269|PubMed:2498334};
CC KM=150 uM for pyruvate {ECO:0000269|PubMed:2498334};
CC KM=6.6 uM for NADPH {ECO:0000269|PubMed:2498334};
CC Vmax=7.9 umol/min/mg enzyme with L-ornithine as substrate
CC {ECO:0000269|PubMed:2498334};
CC Vmax=5.9 umol/min/mg enzyme with L-lysine as substrate
CC {ECO:0000269|PubMed:2498334};
CC pH dependence:
CC Optimum pH is 8.0. Active over a broad pH range of 6.5-9.0.
CC {ECO:0000269|PubMed:2498334};
CC Temperature dependence:
CC Stable to heating at 50 degrees Celsius for 10 minutes. Rapidly
CC inactivated by heating at 60 degrees Celsius.
CC {ECO:0000269|PubMed:2498334};
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:10548058}.
CC -!- MASS SPECTROMETRY: Mass=35355; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:10548058};
CC -!- MISCELLANEOUS: No enzyme activity is detectable in the reverse
CC direction with NADP(+) and chemically synthesized N(5)-(L-1-
CC carboxyethyl)-L-ornithine as potential substrates.
CC -!- SIMILARITY: Belongs to the AlaDH/PNT family. CEOS subfamily.
CC {ECO:0000305}.
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DR EMBL; U23376; AAA86385.1; -; Genomic_DNA.
DR PIR; A57499; A57499.
DR AlphaFoldDB; P15244; -.
DR SMR; P15244; -.
DR SABIO-RK; P15244; -.
DR GO; GO:0047126; F:N5-(carboxyethyl)ornithine synthase activity; IDA:UniProtKB.
DR GO; GO:0006591; P:ornithine metabolic process; IDA:UniProtKB.
DR InterPro; IPR007886; AlaDH/PNT_N.
DR InterPro; IPR007698; AlaDH/PNT_NAD(H)-bd.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF01262; AlaDh_PNT_C; 1.
DR Pfam; PF05222; AlaDh_PNT_N; 1.
DR SMART; SM01002; AlaDh_PNT_C; 1.
DR SMART; SM01003; AlaDh_PNT_N; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; NADP; Oxidoreductase.
FT CHAIN 1..313
FT /note="N(5)-(carboxyethyl)ornithine synthase"
FT /id="PRO_0000089483"
FT BINDING 15
FT /ligand="pyruvate"
FT /ligand_id="ChEBI:CHEBI:15361"
FT /evidence="ECO:0000250"
FT BINDING 71
FT /ligand="pyruvate"
FT /ligand_id="ChEBI:CHEBI:15361"
FT /evidence="ECO:0000250"
FT BINDING 92
FT /ligand="pyruvate"
FT /ligand_id="ChEBI:CHEBI:15361"
FT /evidence="ECO:0000250"
FT BINDING 171..176
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255"
FT MUTAGEN 15
FT /note="R->K: Loss of activity."
FT /evidence="ECO:0000269|PubMed:7744873"
SQ SEQUENCE 313 AA; 35323 MW; B17FE0F477113C77 CRC64;
MKIGLVKANF PGERRVPLLP KDIKDFKNEI LVEEGFGKFL DIDDQEYSDK GCHILSRAEV
FAESEAIFSL KLIQPTDYYH LREGQMIIGW THPFGSGQSF MKEQALPKKL IVVDLDSNSP
CIYYENEIFE SGIPKGLLYK NSFYAGYAGV LDALLQYGLI PTEETKIAIL GSGNVAQGAF
SSISKYSSNI RMYYRKTMSI FKENYTKYDI IINGIEIGKD DDPILSFSEQ KSLKKGTLII
DVAADAGNTI EGSHFTSIDA PIYENAGKYY YVVPNTPSLI YRNVSQELSK ILSENIFRKD
CSRFIEKVKP LNK
