ID DHA_LYSSH Reviewed; 372 AA.
AC P17556;
DT 01-AUG-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1990, sequence version 1.
DT 27-MAR-2024, entry version 116.
DE RecName: Full=Alanine dehydrogenase {ECO:0000303|PubMed:488097};
DE EC=1.4.1.1 {ECO:0000269|PubMed:488097};
GN Name=ald;
OS Lysinibacillus sphaericus (Bacillus sphaericus).
OC Bacteria; Bacillota; Bacilli; Bacillales; Bacillaceae; Lysinibacillus.
OX NCBI_TaxID=1421;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-23; 64-73;
RP 112-151; 157-167; 204-205; 214-230; 281-321; 329-331 AND 343-372, FUNCTION
RP AS AN ALANINE DEHYDROGENASE, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=ATCC 10208 / DSM 5019 / NBRC 3525 / NCIMB 11935 / NRS 966 / 1911;
RX PubMed=2340274; DOI=10.1021/bi00456a025;
RA Kuroda S., Tanizawa K., Sakamoto Y., Tanaka H., Soda K.;
RT "Alanine dehydrogenases from two Bacillus species with distinct
RT thermostabilities: molecular cloning, DNA and protein sequence
RT determination, and structural comparison with other NAD(P)(+)-dependent
RT dehydrogenases.";
RL Biochemistry 29:1009-1015(1990).
RN [2]
RP PROTEIN SEQUENCE OF 1 AND 372, FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE
RP SPECIFICITY, REACTION MECHANISM, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RC STRAIN=ATCC 10208 / DSM 5019 / NBRC 3525 / NCIMB 11935 / NRS 966 / 1911;
RX PubMed=488097; DOI=10.1111/j.1432-1033.1979.tb02030.x;
RA Ohashima T., Soda K.;
RT "Purification and properties of alanine dehydrogenase from Bacillus
RT sphaericus.";
RL Eur. J. Biochem. 100:29-30(1979).
CC -!- FUNCTION: Catalyzes the reversible reductive amination of pyruvate to
CC L-alanine. Prefers L-alanine for oxidative deamination, other
CC substrates are poorly reactive. In the other direction 2-oxobutyrate is
CC almost as reactive as pyruvate. Ammonia is the sole amino donor for the
CC reductive amination of pyruvate, NADPH is inert. Reductive amination
CC proceeds through a sequential, ordered ternary-binary mechanism, where
CC NADH binds first followed by ammonia and pyruvate; the products are
CC released in the order L-alanine and NAD(+) (PubMed:488097,
CC PubMed:2340274). A key factor in the assimilation of L-alanine as an
CC energy source via the tricarboxylic acid cycle during sporulation (By
CC similarity). {ECO:0000250|UniProtKB:Q08352, ECO:0000269|PubMed:2340274,
CC ECO:0000269|PubMed:488097}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-alanine + NAD(+) = H(+) + NADH + NH4(+) + pyruvate;
CC Xref=Rhea:RHEA:18405, ChEBI:CHEBI:15361, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:28938, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945, ChEBI:CHEBI:57972; EC=1.4.1.1;
CC Evidence={ECO:0000269|PubMed:488097};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:18406;
CC Evidence={ECO:0000269|PubMed:488097};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:18407;
CC Evidence={ECO:0000269|PubMed:488097};
CC -!- ACTIVITY REGULATION: Inhibited by p-chloromercuribenzoate and HgCl(2)
CC and by Cu(2+) and Pb(2+) salts, unaffected by amino acids such as D-
CC alanine and beta-alanine or by nucleotides or nucleosides.
CC {ECO:0000269|PubMed:488097}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=18.9 uM for oxidative deamination of L-alanine
CC {ECO:0000269|PubMed:488097};
CC KM=330 uM for oxidative deamination of L-2-aminobutyrate
CC {ECO:0000269|PubMed:488097};
CC KM=39 uM for oxidative deamination of L-serine
CC {ECO:0000269|PubMed:488097};
CC KM=20 uM for oxidative deamination of L-valine
CC {ECO:0000269|PubMed:488097};
CC KM=14 uM for oxidative deamination of L-norvaline
CC {ECO:0000269|PubMed:488097};
CC KM=1.7 mM for reductive amination of pyruvate
CC {ECO:0000269|PubMed:488097};
CC KM=80 mM for reductive amination of 3-hydroxypyruvate
CC {ECO:0000269|PubMed:488097};
CC KM=23 mM for reductive amination of 2-oxovalerate
CC {ECO:0000269|PubMed:488097};
CC KM=12 mM for reductive amination of glyoxylate
CC {ECO:0000269|PubMed:488097};
CC KM=11 mM for reductive amination of 2-oxobutyrate
CC {ECO:0000269|PubMed:488097};
CC KM=11 mM for reductive amination of 2-oxo-3-methylbutanoate
CC {ECO:0000269|PubMed:488097};
CC pH dependence:
CC Optimum pH is 10.0-10.5 for the oxidative deamination of L-alanine
CC and about 9.0 for reductive amination of pyruvate.
CC {ECO:0000269|PubMed:488097};
CC Temperature dependence:
CC Loses about 50% of its initial activity when heated at 65 degrees
CC Celsius for 5 min. {ECO:0000269|PubMed:2340274,
CC ECO:0000269|PubMed:488097};
CC -!- PATHWAY: Amino-acid degradation; L-alanine degradation via
CC dehydrogenase pathway; NH(3) and pyruvate from L-alanine: step 1/1.
CC -!- SUBUNIT: Homohexamer. {ECO:0000269|PubMed:2340274,
CC ECO:0000269|PubMed:488097}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250|UniProtKB:A0QVQ8}.
CC -!- SIMILARITY: Belongs to the AlaDH/PNT family. {ECO:0000305}.
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DR EMBL; M33298; AAA22210.1; -; Genomic_DNA.
DR PIR; A34261; A34261.
DR AlphaFoldDB; P17556; -.
DR SMR; P17556; -.
DR STRING; 1421.A2J09_13020; -.
DR UniPathway; UPA00527; UER00585.
DR GO; GO:0005829; C:cytosol; ISS:UniProtKB.
DR GO; GO:0000286; F:alanine dehydrogenase activity; IDA:UniProtKB.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0006524; P:alanine catabolic process; ISS:UniProtKB.
DR GO; GO:0042853; P:L-alanine catabolic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0030435; P:sporulation resulting in formation of a cellular spore; IEA:UniProtKB-KW.
DR CDD; cd05305; L-AlaDH; 1.
DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 2.
DR InterPro; IPR008141; Ala_DH.
DR InterPro; IPR008143; Ala_DH/PNT_CS2.
DR InterPro; IPR008142; AlaDH/PNT_CS1.
DR InterPro; IPR007886; AlaDH/PNT_N.
DR InterPro; IPR007698; AlaDH/PNT_NAD(H)-bd.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR NCBIfam; TIGR00518; alaDH; 1.
DR PANTHER; PTHR42795; ALANINE DEHYDROGENASE; 1.
DR PANTHER; PTHR42795:SF1; ALANINE DEHYDROGENASE 1; 1.
DR Pfam; PF01262; AlaDh_PNT_C; 1.
DR Pfam; PF05222; AlaDh_PNT_N; 1.
DR PIRSF; PIRSF000183; Alanine_dh; 1.
DR SMART; SM01002; AlaDh_PNT_C; 1.
DR SMART; SM01003; AlaDh_PNT_N; 1.
DR SUPFAM; SSF52283; Formate/glycerate dehydrogenase catalytic domain-like; 1.
DR SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
DR PROSITE; PS00836; ALADH_PNT_1; 1.
DR PROSITE; PS00837; ALADH_PNT_2; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Direct protein sequencing; NAD; Nucleotide-binding;
KW Oxidoreductase; Sporulation.
FT CHAIN 1..372
FT /note="Alanine dehydrogenase"
FT /id="PRO_0000198990"
FT ACT_SITE 96
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000255"
FT ACT_SITE 270
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250"
FT BINDING 15
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 75
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 134
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 178..179
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 198
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 220
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 239..240
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 267..270
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 280
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 299..302
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
SQ SEQUENCE 372 AA; 39460 MW; 5E6FD5E4D93AA98B CRC64;
MKIGIPKEIK NNENRVAMTP AGVVSLTHAG HERLAIETGG GIGSSFTDAE YVAAGAAYRC
IGKEAWAQEM ILKVKEPVAS EYDYFYEGQI LFTYLHLAPR AELTQALIDK KVVGIAYETV
QLANGSLPLL TPMSEVAGKM ATQIGAQYLE KNHGGKGILL GGVSGVHARK VTVIGGGIAG
TNAAKIAVGM GADVTVIDLS PERLRQLEDM FGRDVQTLMS NPYNIAESVK HSDLVVGAVL
IPGAKAPKLV SEEMIQSMQP GSVVVDIAID QGGIFATSDR VTTHDDPTYV KHGVVHYAVA
NMPGAVPRTS TIALTNNTIP YALQIANKGY KQACIDNPAL KKGVNALEGH ITYKAVAEAQ
GLPYVNVDEL IQ
//
