ID   DHLE_GEOSE              Reviewed;         367 AA.
AC   P13154; Q76GN7;
DT   01-JAN-1990, integrated into UniProtKB/Swiss-Prot.
DT   25-OCT-2004, sequence version 3.
DT   03-AUG-2022, entry version 113.
DE   RecName: Full=Leucine dehydrogenase;
DE            Short=LeuDH;
DE            EC=1.4.1.9;
GN   Name=ldh; Synonyms=leuDH;
OS   Geobacillus stearothermophilus (Bacillus stearothermophilus).
OC   Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Geobacillus.
OX   NCBI_TaxID=1422;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], PARTIAL PROTEIN SEQUENCE, FUNCTION,
RP   CATALYTIC ACTIVITY, AND SUBUNIT.
RC   STRAIN=ATCC 12980 / DSM 22 / CCM 2062 / JCM 2501 / NBRC 12550 / NCIMB 8923
RC   / NCTC 10339 / R-35646 / VKM B-510;
RX   PubMed=3069133; DOI=10.1021/bi00425a026;
RA   Nagata S., Tanizawa K., Esaki N., Sakamoto Y., Ohshima T., Tanaka H.,
RA   Soda K.;
RT   "Gene cloning and sequence determination of leucine dehydrogenase from
RT   Bacillus stearothermophilus and structural comparison with other NAD(P)+-
RT   dependent dehydrogenases.";
RL   Biochemistry 27:9056-9062(1988).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA   Katoh R., Seki M., Nagata S., Misono H.;
RT   "Nucleotide sequence, cloning, overexpression and site-directed mutagenesis
RT   of the leucine dehydrogenase gene from Bacillus sphaericus.";
RL   Submitted (FEB-2003) to the EMBL/GenBank/DDBJ databases.
RN   [3]
RP   ACTIVE SITE LYS-80.
RX   PubMed=1400267; DOI=10.1093/oxfordjournals.jbchem.a123887;
RA   Matsuyama T., Soda K., Fukui T., Tanizawa K.;
RT   "Leucine dehydrogenase from Bacillus stearothermophilus: identification of
RT   active-site lysine by modification with pyridoxal phosphate.";
RL   J. Biochem. 112:258-265(1992).
RN   [4]
RP   MUTAGENESIS OF LYS-68; GLU-114; ASP-261 AND VAL-291, AND BIOTECHNOLOGY.
RX   PubMed=22396126; DOI=10.1002/anie.201107813;
RA   Abrahamson M.J., Vazquez-Figueroa E., Woodall N.B., Moore J.C.,
RA   Bommarius A.S.;
RT   "Development of an amine dehydrogenase for synthesis of chiral amines.";
RL   Angew. Chem. Int. Ed. 51:3969-3972(2012).
CC   -!- FUNCTION: Catalyzes the reversible deamination of L-leucine to 4-
CC       methyl-2-oxopentanoate. {ECO:0000269|PubMed:3069133}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=H2O + L-leucine + NAD(+) = 4-methyl-2-oxopentanoate + H(+) +
CC         NADH + NH4(+); Xref=Rhea:RHEA:12220, ChEBI:CHEBI:15377,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:17865, ChEBI:CHEBI:28938,
CC         ChEBI:CHEBI:57427, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; EC=1.4.1.9;
CC         Evidence={ECO:0000269|PubMed:3069133};
CC   -!- PATHWAY: Amino-acid degradation; L-leucine degradation; 4-methyl-2-
CC       oxopentanoate from L-leucine (dehydrogenase route): step 1/1.
CC   -!- SUBUNIT: Homohexamer. {ECO:0000269|PubMed:3069133}.
CC   -!- BIOTECHNOLOGY: This enzyme has been successfully altered through
CC       several rounds of protein engineering to an enantioselective amine
CC       dehydrogenase. Instead of the wild-type alpha-keto acid, the new amine
CC       dehydrogenase now accepts the analogous ketone, methyl isobutyl ketone
CC       (MIBK), which corresponds to exchange of the carboxy group by a methyl
CC       group, to produce chiral (R)-1,3-dimethylbutylamine via a reductive
CC       amination reaction. This represents a suitable enzymatic production
CC       route for the asymmetric synthesis of amines from prochiral ketones and
CC       free ammonia, which is one of the top aspirational reactions
CC       challenging the pharmaceutical industry. {ECO:0000269|PubMed:22396126}.
CC   -!- SIMILARITY: Belongs to the Glu/Leu/Phe/Val dehydrogenases family.
CC       {ECO:0000305}.
CC   -!- SEQUENCE CAUTION:
CC       Sequence=AAA22570.1; Type=Frameshift; Evidence={ECO:0000305};
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DR   EMBL; M22977; AAA22570.1; ALT_FRAME; Genomic_DNA.
DR   EMBL; AB103384; BAC81829.1; -; Genomic_DNA.
DR   PIR; A31950; A31950.
DR   AlphaFoldDB; P13154; -.
DR   SMR; P13154; -.
DR   BRENDA; 1.4.1.9; 623.
DR   UniPathway; UPA00363; UER00858.
DR   GO; GO:0050049; F:leucine dehydrogenase activity; IEA:UniProtKB-EC.
DR   GO; GO:0006552; P:leucine catabolic process; IEA:UniProtKB-UniPathway.
DR   InterPro; IPR046346; Aminiacid_DH-like_N_sf.
DR   InterPro; IPR006095; Glu/Leu/Phe/Val_DH.
DR   InterPro; IPR033524; Glu/Leu/Phe/Val_DH_AS.
DR   InterPro; IPR006096; Glu/Leu/Phe/Val_DH_C.
DR   InterPro; IPR006097; Glu/Leu/Phe/Val_DH_dimer_dom.
DR   InterPro; IPR016211; Glu/Phe/Leu/Val_DH_bac/arc.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   PANTHER; PTHR42722; PTHR42722; 1.
DR   Pfam; PF00208; ELFV_dehydrog; 2.
DR   Pfam; PF02812; ELFV_dehydrog_N; 1.
DR   PIRSF; PIRSF000188; Phe_leu_dh; 1.
DR   PRINTS; PR00082; GLFDHDRGNASE.
DR   SMART; SM00839; ELFV_dehydrog; 1.
DR   SUPFAM; SSF51735; SSF51735; 1.
DR   SUPFAM; SSF53223; SSF53223; 1.
DR   PROSITE; PS00074; GLFV_DEHYDROGENASE; 1.
PE   1: Evidence at protein level;
KW   Branched-chain amino acid catabolism; Direct protein sequencing; NAD;
KW   Oxidoreductase.
FT   CHAIN           1..367
FT                   /note="Leucine dehydrogenase"
FT                   /id="PRO_0000182803"
FT   ACT_SITE        80
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU10011,
FT                   ECO:0000269|PubMed:1400267"
FT   BINDING         180..186
FT                   /ligand="NAD(+)"
FT                   /ligand_id="ChEBI:CHEBI:57540"
FT                   /evidence="ECO:0000255"
FT   MUTAGEN         68
FT                   /note="K->S: Completely modifies the activity of the
FT                   enzyme, which is able to catalyze the reductive amination
FT                   of ketones, and no longer displays leucine dehydrogenase
FT                   activity; when associated with V-114, C-261 and C-291."
FT                   /evidence="ECO:0000269|PubMed:22396126"
FT   MUTAGEN         114
FT                   /note="E->V: Completely modifies the activity of the
FT                   enzyme, which is able to catalyze the reductive amination
FT                   of ketones, and no longer displays leucine dehydrogenase
FT                   activity; when associated with S-68, C-261 and C-291."
FT                   /evidence="ECO:0000269|PubMed:22396126"
FT   MUTAGEN         261
FT                   /note="D->C: Completely modifies the activity of the
FT                   enzyme, which is able to catalyze the reductive amination
FT                   of ketones, and no longer displays leucine dehydrogenase
FT                   activity; when associated with S-68, V-114 and C-291."
FT                   /evidence="ECO:0000269|PubMed:22396126"
FT   MUTAGEN         291
FT                   /note="V->C: Completely modifies the activity of the
FT                   enzyme, which is able to catalyze the reductive amination
FT                   of ketones, and no longer displays leucine dehydrogenase
FT                   activity; when associated with S-68, V-114 and C-261."
FT                   /evidence="ECO:0000269|PubMed:22396126"
FT   CONFLICT        350..352
FT                   /note="RSP -> ASQ (in Ref. 1; AAA22570)"
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   367 AA;  40458 MW;  E01E7086453D33DD CRC64;
     MELFKYMETY DYEQVLFCQD KESGLKAIIA IHDTTLGPAL GGTRMWMYNS EEEALEDALR
     LARGMTYKNA AAGLNLGGGK TVIIGDPRKD KNEAMFRAFG RFIQGLNGRY ITAEDVGTTV
     ADMDIIYQET DYVTGISPEF GSSGNPSPAT AYGVYRGMKA AAKEAFGSDS LEGKVVAVQG
     VGNVAYHLCR HLHEEGAKLI VTDINKEVVA RAVEEFGAKA VDPNDIYGVE CDIFAPCALG
     GIINDQTIPQ LKAKVIAGSA DNQLKEPRHG DIIHEMGIVY APDYVINAGG VINVADELYG
     YNRERAMKKI EQIYDNIEKV FAIAKRDNIP TYVAADRMAE ERIETMRKAR SPFLQNGHHI
     LSRRRAR