ID   MAO4_BACSU              Reviewed;         410 AA.
AC   O34962;
DT   28-FEB-2003, integrated into UniProtKB/Swiss-Prot.
DT   01-JAN-1998, sequence version 1.
DT   27-MAR-2024, entry version 145.
DE   RecName: Full=Bifunctional malic/malolactic enzyme {ECO:0000303|PubMed:33824210};
DE            EC=1.1.1.40 {ECO:0000269|PubMed:16788182, ECO:0000269|PubMed:33824210};
DE            EC=4.1.1.101 {ECO:0000269|PubMed:33824210};
DE   AltName: Full=Malolactic enzyme;
DE            Short=MLE;
DE   AltName: Full=NADP-dependent malic enzyme;
DE            Short=NADP-ME;
GN   Name=ytsJ; OrderedLocusNames=BSU29220;
OS   Bacillus subtilis (strain 168).
OC   Bacteria; Bacillota; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX   NCBI_TaxID=224308;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9387221; DOI=10.1099/00221287-143-11-3431;
RA   Lapidus A., Galleron N., Sorokin A., Ehrlich S.D.;
RT   "Sequencing and functional annotation of the Bacillus subtilis genes in the
RT   200 kb rrnB-dnaB region.";
RL   Microbiology 143:3431-3441(1997).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9384377; DOI=10.1038/36786;
RA   Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA   Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA   Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA   Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA   Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA   Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA   Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA   Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA   Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA   Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA   Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA   Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA   Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA   Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA   Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA   Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA   Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA   Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA   Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA   Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA   Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA   Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA   Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA   Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA   Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA   Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA   Yoshikawa H., Danchin A.;
RT   "The complete genome sequence of the Gram-positive bacterium Bacillus
RT   subtilis.";
RL   Nature 390:249-256(1997).
RN   [3]
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, INDUCTION, AND
RP   DISRUPTION PHENOTYPE.
RC   STRAIN=168;
RX   PubMed=16788182; DOI=10.1128/jb.00167-06;
RA   Lerondel G., Doan T., Zamboni N., Sauer U., Aymerich S.;
RT   "YtsJ has the major physiological role of the four paralogous malic enzyme
RT   isoforms in Bacillus subtilis.";
RL   J. Bacteriol. 188:4727-4736(2006).
RN   [4]
RP   DISRUPTION PHENOTYPE.
RC   STRAIN=168;
RX   PubMed=23136871; DOI=10.1111/1574-6968.12041;
RA   Meyer F.M., Stuelke J.;
RT   "Malate metabolism in Bacillus subtilis: distinct roles for three classes
RT   of malate-oxidizing enzymes.";
RL   FEMS Microbiol. Lett. 339:17-22(2013).
RN   [5]
RP   FUNCTION AS A BIFUNCTIONAL ENZYME, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP   BIOPHYSICOCHEMICAL PROPERTIES, AND DISRUPTION PHENOTYPE.
RX   PubMed=33824210; DOI=10.1128/mbio.03438-20;
RA   Hoerl M., Fuhrer T., Zamboni N.;
RT   "Bifunctional malic/malolactic enzyme provides a novel mechanism for NADPH-
RT   balancing in Bacillus subtilis.";
RL   MBio 12:0-0(2021).
RN   [6]
RP   INTERACTION WITH BRXC, AND IDENTIFICATION BY MASS SPECTROMETRY.
RC   STRAIN=168 / CU1065 {ECO:0000303|PubMed:33722570};
RX   PubMed=33722570; DOI=10.1016/j.redox.2021.101935;
RA   Gaballa A., Su T.T., Helmann J.D.;
RT   "The Bacillus subtilis monothiol bacilliredoxin BrxC (YtxJ) and the Bdr
RT   (YpdA) disulfide reductase reduce S-bacillithiolated proteins.";
RL   Redox Biol. 42:101935-101935(2021).
CC   -!- FUNCTION: Bifunctional enzyme with both malic and malolactic enzyme
CC       activities (PubMed:33824210). In the absence of NADPH, catalyzes the
CC       reversible decarboxylation of malate to pyruvate (PubMed:16788182,
CC       PubMed:33824210). Can use NAD and NADP, but with a very strong
CC       preference for NADP (PubMed:16788182). In the presence of excess NADPH,
CC       catalyzes the non-oxidative decarboxylation of malate to lactate
CC       (PubMed:33824210). During growth on glucose, contributes to NADPH
CC       balancing via oxidation of the NADPH produced in excess by other
CC       enzymatic reactions (PubMed:33824210). Can also catalyze the
CC       decarboxylation of oxaloacetate (PubMed:16788182).
CC       {ECO:0000269|PubMed:16788182, ECO:0000269|PubMed:33824210}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-malate + NADP(+) = CO2 + NADPH + pyruvate;
CC         Xref=Rhea:RHEA:18253, ChEBI:CHEBI:15361, ChEBI:CHEBI:15589,
CC         ChEBI:CHEBI:16526, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.1.1.40;
CC         Evidence={ECO:0000269|PubMed:16788182, ECO:0000269|PubMed:33824210};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=H(+) + oxaloacetate = CO2 + pyruvate; Xref=Rhea:RHEA:15641,
CC         ChEBI:CHEBI:15361, ChEBI:CHEBI:15378, ChEBI:CHEBI:16452,
CC         ChEBI:CHEBI:16526; EC=1.1.1.40;
CC         Evidence={ECO:0000269|PubMed:16788182};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-malate + H(+) = (S)-lactate + CO2; Xref=Rhea:RHEA:46276,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:15589, ChEBI:CHEBI:16526,
CC         ChEBI:CHEBI:16651; EC=4.1.1.101;
CC         Evidence={ECO:0000269|PubMed:33824210};
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC         Evidence={ECO:0000250|UniProtKB:P76558};
CC       Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC         Evidence={ECO:0000250|UniProtKB:P76558};
CC       Note=Divalent metal cations. {ECO:0000250|UniProtKB:P76558};
CC   -!- ACTIVITY REGULATION: NADPH is a strong modulator that switches activity
CC       from a pyruvate-producing malic enzyme to a lactate-generating
CC       malolactic enzyme. {ECO:0000269|PubMed:33824210}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=1.55 mM for malate {ECO:0000269|PubMed:16788182};
CC         KM=5.3 mM for pyruvate {ECO:0000269|PubMed:33824210};
CC         KM=2.8 mM for NAD {ECO:0000269|PubMed:16788182};
CC         KM=0.055 mM for NADP {ECO:0000269|PubMed:16788182};
CC         KM=0.8 mM for NADPH {ECO:0000269|PubMed:33824210};
CC   -!- SUBUNIT: Interacts with BrxC. {ECO:0000269|PubMed:33722570}.
CC   -!- INDUCTION: Transcribed at a high level under all of the conditions
CC       tested. {ECO:0000269|PubMed:16788182}.
CC   -!- DISRUPTION PHENOTYPE: Mutant shows wild-type growth on glucose but a
CC       much slower growth rate on malate, fumarate, or succinate plus
CC       glutamate (PubMed:16788182). Overexpression of the other three malic
CC       enzymes, mleA, maeA or malS cannot compensate for the ytsJ deletion
CC       (PubMed:16788182). The ATP concentrations in the mutant grown in
CC       minimal medium with glucose are similar to the wild-type level. ATP
CC       concentrations decrease by about 20% in malate minimal medium
CC       (PubMed:23136871). NADPH overproduction is virtually abolished in the
CC       deletion mutant (PubMed:33824210). {ECO:0000269|PubMed:16788182,
CC       ECO:0000269|PubMed:23136871, ECO:0000269|PubMed:33824210}.
CC   -!- SIMILARITY: Belongs to the malic enzymes family. {ECO:0000305}.
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DR   EMBL; AF008220; AAC00339.1; -; Genomic_DNA.
DR   EMBL; AL009126; CAB14882.1; -; Genomic_DNA.
DR   PIR; C70001; C70001.
DR   RefSeq; NP_390800.1; NC_000964.3.
DR   RefSeq; WP_003229415.1; NZ_JNCM01000036.1.
DR   AlphaFoldDB; O34962; -.
DR   SMR; O34962; -.
DR   IntAct; O34962; 2.
DR   MINT; O34962; -.
DR   STRING; 224308.BSU29220; -.
DR   jPOST; O34962; -.
DR   PaxDb; 224308-BSU29220; -.
DR   DNASU; 937378; -.
DR   EnsemblBacteria; CAB14882; CAB14882; BSU_29220.
DR   GeneID; 937378; -.
DR   KEGG; bsu:BSU29220; -.
DR   PATRIC; fig|224308.179.peg.3173; -.
DR   eggNOG; COG0281; Bacteria.
DR   InParanoid; O34962; -.
DR   OrthoDB; 9805787at2; -.
DR   PhylomeDB; O34962; -.
DR   BioCyc; BSUB:BSU29220-MONOMER; -.
DR   Proteomes; UP000001570; Chromosome.
DR   GO; GO:0004471; F:malate dehydrogenase (decarboxylating) (NAD+) activity; IEA:UniProtKB-EC.
DR   GO; GO:0004473; F:malate dehydrogenase (decarboxylating) (NADP+) activity; IEA:RHEA.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   GO; GO:0051287; F:NAD binding; IEA:InterPro.
DR   GO; GO:0008948; F:oxaloacetate decarboxylase activity; IEA:UniProtKB-EC.
DR   CDD; cd05311; NAD_bind_2_malic_enz; 1.
DR   Gene3D; 3.40.50.10380; Malic enzyme, N-terminal domain; 1.
DR   Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1.
DR   InterPro; IPR046346; Aminoacid_DH-like_N_sf.
DR   InterPro; IPR015884; Malic_enzyme_CS.
DR   InterPro; IPR012301; Malic_N_dom.
DR   InterPro; IPR037062; Malic_N_dom_sf.
DR   InterPro; IPR012302; Malic_NAD-bd.
DR   InterPro; IPR045213; Malic_NAD-bd_bact_type.
DR   InterPro; IPR001891; Malic_OxRdtase.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   PANTHER; PTHR43237; NADP-DEPENDENT MALIC ENZYME; 1.
DR   PANTHER; PTHR43237:SF4; NADP-DEPENDENT MALIC ENZYME; 1.
DR   Pfam; PF00390; malic; 1.
DR   Pfam; PF03949; Malic_M; 1.
DR   PIRSF; PIRSF000106; ME; 1.
DR   PRINTS; PR00072; MALOXRDTASE.
DR   SMART; SM01274; malic; 1.
DR   SMART; SM00919; Malic_M; 1.
DR   SUPFAM; SSF53223; Aminoacid dehydrogenase-like, N-terminal domain; 1.
DR   SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
DR   PROSITE; PS00331; MALIC_ENZYMES; 1.
PE   1: Evidence at protein level;
KW   Lyase; Magnesium; Manganese; Metal-binding; NADP; Oxidoreductase;
KW   Reference proteome.
FT   CHAIN           1..410
FT                   /note="Bifunctional malic/malolactic enzyme"
FT                   /id="PRO_0000160211"
FT   ACT_SITE        36
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   ACT_SITE        91
FT                   /note="Proton acceptor"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   BINDING         133
FT                   /ligand="a divalent metal cation"
FT                   /ligand_id="ChEBI:CHEBI:60240"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   BINDING         134
FT                   /ligand="a divalent metal cation"
FT                   /ligand_id="ChEBI:CHEBI:60240"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   BINDING         159
FT                   /ligand="a divalent metal cation"
FT                   /ligand_id="ChEBI:CHEBI:60240"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   BINDING         192..195
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   BINDING         286
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
FT   BINDING         317
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000250|UniProtKB:P40927"
SQ   SEQUENCE   410 AA;  43667 MW;  3B9B2FDAAA349D61 CRC64;
     MSLREEALHL HKVNQGKLES KSKVEVRNAK DLSLAYSPGV AEPCKDIHED INKVYDYTMK
     GNMVAVVTDG TAVLGLGNIG PEAALPVMEG KAVLFKSFAG VDAFPIALNT NDVDKIVETV
     KLLEPTFGGV NLEDIAAPNC FIIEERLKKE TNIPVFHDDQ HGTAIVTVAG LVNALKLSGK
     SMSSIKVVAN GAGAAGIAII KLLHHYGVRD IVMCDSKGAI YEGRPNGMND VKNEVAKFTN
     QDRKDGSLKD VIVDADVFIG VSVAGALTKE MVQSMAKDPI IFAMANPNPE IMPEDAREAG
     ASVVGTGRSD FPNQVNNVLA FPGIFRGALD VRATHINEEM KIAAVEAIAS LVSEDELSAD
     YVIPAPFDKR VAPAVAKAVA KAAMETGVAR ITVDPEEVAE KTRKLTIIGE
//