ID CAR_MYCMM Reviewed; 1174 AA.
AC B2HN69;
DT 19-FEB-2014, integrated into UniProtKB/Swiss-Prot.
DT 10-JUN-2008, sequence version 1.
DT 27-NOV-2024, entry version 91.
DE RecName: Full=Carboxylic acid reductase {ECO:0000255|HAMAP-Rule:MF_02247, ECO:0000303|PubMed:23248280};
DE Short=CAR {ECO:0000255|HAMAP-Rule:MF_02247, ECO:0000303|PubMed:23248280};
DE EC=1.2.1.- {ECO:0000255|HAMAP-Rule:MF_02247};
DE AltName: Full=ATP/NADPH-dependent carboxylic acid reductase {ECO:0000255|HAMAP-Rule:MF_02247};
DE AltName: Full=Fatty acid reductase;
GN Name=car {ECO:0000255|HAMAP-Rule:MF_02247}; Synonyms=fadD9;
GN OrderedLocusNames=MMAR_2117;
OS Mycobacterium marinum (strain ATCC BAA-535 / M).
OC Bacteria; Actinomycetota; Actinomycetes; Mycobacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium ulcerans group.
OX NCBI_TaxID=216594;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC BAA-535 / M;
RX PubMed=18403782; DOI=10.1101/gr.075069.107;
RA Stinear T.P., Seemann T., Harrison P.F., Jenkin G.A., Davies J.K.,
RA Johnson P.D., Abdellah Z., Arrowsmith C., Chillingworth T., Churcher C.,
RA Clarke K., Cronin A., Davis P., Goodhead I., Holroyd N., Jagels K.,
RA Lord A., Moule S., Mungall K., Norbertczak H., Quail M.A.,
RA Rabbinowitsch E., Walker D., White B., Whitehead S., Small P.L., Brosch R.,
RA Ramakrishnan L., Fischbach M.A., Parkhill J., Cole S.T.;
RT "Insights from the complete genome sequence of Mycobacterium marinum on the
RT evolution of Mycobacterium tuberculosis.";
RL Genome Res. 18:729-741(2008).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, COFACTOR, AND BIOTECHNOLOGY.
RX PubMed=23248280; DOI=10.1073/pnas.1216516110;
RA Akhtar M.K., Turner N.J., Jones P.R.;
RT "Carboxylic acid reductase is a versatile enzyme for the conversion of
RT fatty acids into fuels and chemical commodities.";
RL Proc. Natl. Acad. Sci. U.S.A. 110:87-92(2013).
RN [3] {ECO:0007744|PDB:5MSO, ECO:0007744|PDB:5MSU}
RP X-RAY CRYSTALLOGRAPHY (1.20 ANGSTROMS) IN COMPLEX WITH NADP, AND DOMAIN.
RX PubMed=28719588; DOI=10.1038/nchembio.2434;
RA Gahloth D., Dunstan M.S., Quaglia D., Klumbys E., Lockhart-Cairns M.P.,
RA Hill A.M., Derrington S.R., Scrutton N.S., Turner N.J., Leys D.;
RT "Structures of carboxylic acid reductase reveal domain dynamics underlying
RT catalysis.";
RL Nat. Chem. Biol. 13:975-981(2017).
CC -!- FUNCTION: Catalyzes the ATP- and NADPH-dependent reduction of
CC carboxylic acids to the corresponding aldehydes (PubMed:23248280).
CC Catalyzes the reduction of a wide range of aliphatic fatty acids (C6-
CC C18) into their corresponding aldehydes. Can also reduce benzoate to
CC benzaldehyde. Has a preference for NADPH over NADH as the electron
CC donor (PubMed:23248280). {ECO:0000269|PubMed:23248280}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a carboxylate + ATP + NADPH + H(+) = an aldehyde + AMP +
CC diphosphate + NADP(+); Xref=Rhea:RHEA:50916, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17478, ChEBI:CHEBI:29067, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:456215; Evidence={ECO:0000255|HAMAP-Rule:MF_02247,
CC ECO:0000269|PubMed:23248280};
CC -!- COFACTOR:
CC Name=pantetheine 4'-phosphate; Xref=ChEBI:CHEBI:47942;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_02247,
CC ECO:0000305|PubMed:23248280};
CC Note=Binds 1 phosphopantetheine covalently. {ECO:0000255|HAMAP-
CC Rule:MF_02247, ECO:0000305|PubMed:23248280};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=362 uM for benzoate {ECO:0000269|PubMed:23248280};
CC KM=48 uM for NADPH {ECO:0000269|PubMed:23248280};
CC KM=115 uM for ATP {ECO:0000269|PubMed:23248280};
CC Vmax=2.32 umol/min/mg enzyme with benzoate as substrate
CC {ECO:0000269|PubMed:23248280};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:23248280};
CC Temperature dependence:
CC Displays in vitro half-lives of 73, 70, and 48 hours at 26, 30, and
CC 37 degrees Celsius, respectively, indicating it is a relatively
CC stable enzyme. {ECO:0000269|PubMed:23248280};
CC -!- DOMAIN: The N-terminal domain likely catalyzes substrate activation by
CC formation of an initial acyl-AMP intermediate, the central region
CC contains the phosphopantetheine attachment site, and the C-terminal
CC domain catalyzes the reduction by NADPH of the intermediate thioester
CC formed from the attack of the phosphopantetheine thiol at the carbonyl
CC carbon of acyl-AMP (By similarity). Large-scale domain motions occur
CC between the adenylation and thiolation states. Phosphopantetheine
CC binding alters the orientation of a key Asp, resulting in a productive
CC orientation of the bound nicotinamide. This ensures that further
CC reduction of the aldehyde product does not occur (PubMed:28719588).
CC {ECO:0000250|UniProtKB:Q6RKB1, ECO:0000269|PubMed:28719588}.
CC -!- BIOTECHNOLOGY: This enzyme can be applied to the microbial production
CC of fatty alkanes and fatty alcohols that have numerous applications as
CC fuels, fragrances, emollients, plasticizers, thickeners, and
CC detergents. Thus, together with complementing enzymes, the broad
CC substrate specificity and kinetic characteristics of Car opens the road
CC for direct and tailored enzyme-catalyzed conversion of lipids into
CC user-ready chemical commodities. {ECO:0000269|PubMed:23248280}.
CC -!- MISCELLANEOUS: The conversion of fatty acid into aldehyde involves
CC three key steps: adenylation of the bound fatty acid substrate to form
CC an AMP-fatty acyl complex, formation of a thioester linkage between the
CC fatty acyl moiety and the phosphopantetheine prosthetic group, and
CC reduction of the thioester intermediate to the aldehyde.
CC {ECO:0000305|PubMed:23248280}.
CC -!- SIMILARITY: Belongs to the ATP-dependent AMP-binding enzyme family.
CC Carboxylic acid reductase subfamily. {ECO:0000255|HAMAP-Rule:MF_02247,
CC ECO:0000305}.
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DR EMBL; CP000854; ACC40567.1; -; Genomic_DNA.
DR RefSeq; WP_012393886.1; NC_010612.1.
DR PDB; 5MSO; X-ray; 1.20 A; A=1-1174.
DR PDB; 5MSU; X-ray; 1.74 A; A/B/C=1-1174.
DR PDBsum; 5MSO; -.
DR PDBsum; 5MSU; -.
DR AlphaFoldDB; B2HN69; -.
DR SMR; B2HN69; -.
DR STRING; 216594.MMAR_2117; -.
DR KEGG; mmi:MMAR_2117; -.
DR eggNOG; COG1022; Bacteria.
DR eggNOG; COG3320; Bacteria.
DR HOGENOM; CLU_009549_0_0_11; -.
DR OrthoDB; 2472181at2; -.
DR BRENDA; 1.2.1.30; 3506.
DR Proteomes; UP000001190; Chromosome.
DR GO; GO:0016020; C:membrane; IEA:TreeGrafter.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0004467; F:long-chain fatty acid-CoA ligase activity; IEA:TreeGrafter.
DR GO; GO:0050661; F:NADP binding; IEA:UniProtKB-UniRule.
DR GO; GO:0016620; F:oxidoreductase activity, acting on the aldehyde or oxo group of donors, NAD or NADP as acceptor; IEA:UniProtKB-UniRule.
DR GO; GO:0031177; F:phosphopantetheine binding; IEA:UniProtKB-UniRule.
DR CDD; cd17632; AFD_CAR-like; 1.
DR CDD; cd05235; SDR_e1; 1.
DR Gene3D; 1.10.1200.10; ACP-like; 1.
DR Gene3D; 3.40.50.12780; N-terminal domain of ligase-like; 1.
DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1.
DR HAMAP; MF_02247; Carbox_acid_reduct; 1.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig_com.
DR InterPro; IPR042099; ANL_N_sf.
DR InterPro; IPR046407; CAR.
DR InterPro; IPR013120; Far_NAD-bd.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR020806; PKS_PP-bd.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR010080; Thioester_reductase-like_dom.
DR NCBIfam; NF041592; carboxyl_red; 1.
DR NCBIfam; TIGR01746; Thioester-redct; 1.
DR PANTHER; PTHR43272:SF52; CARBOXYLIC ACID REDUCTASE; 1.
DR PANTHER; PTHR43272; LONG-CHAIN-FATTY-ACID--COA LIGASE; 1.
DR Pfam; PF00501; AMP-binding; 1.
DR Pfam; PF07993; NAD_binding_4; 1.
DR Pfam; PF00550; PP-binding; 1.
DR SMART; SM00823; PKS_PP; 1.
DR SUPFAM; SSF56801; Acetyl-CoA synthetase-like; 1.
DR SUPFAM; SSF47336; ACP-like; 1.
DR SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
DR PROSITE; PS00061; ADH_SHORT; 1.
DR PROSITE; PS00455; AMP_BINDING; 1.
DR PROSITE; PS50075; CARRIER; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Fatty acid metabolism; Lipid metabolism; NADP;
KW Nucleotide-binding; Oxidoreductase; Phosphopantetheine; Phosphoprotein;
KW Reference proteome.
FT CHAIN 1..1174
FT /note="Carboxylic acid reductase"
FT /id="PRO_0000425448"
FT DOMAIN 651..726
FT /note="Carrier"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 297
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 392
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 413..414
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 418
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 491
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 503..506
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 512
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 612
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT BINDING 787..791
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:28719588,
FT ECO:0007744|PDB:5MSO, ECO:0007744|PDB:5MSU"
FT BINDING 814
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247,
FT ECO:0000269|PubMed:28719588, ECO:0007744|PDB:5MSO,
FT ECO:0007744|PDB:5MSU"
FT BINDING 824
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247,
FT ECO:0000269|PubMed:28719588, ECO:0007744|PDB:5MSO,
FT ECO:0007744|PDB:5MSU"
FT BINDING 854..855
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247,
FT ECO:0000269|PubMed:28719588, ECO:0007744|PDB:5MSO,
FT ECO:0007744|PDB:5MSU"
FT BINDING 880..882
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247,
FT ECO:0000269|PubMed:28719588, ECO:0007744|PDB:5MSO,
FT ECO:0007744|PDB:5MSU"
FT BINDING 919..920
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:28719588,
FT ECO:0007744|PDB:5MSO, ECO:0007744|PDB:5MSU"
FT BINDING 956
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247,
FT ECO:0000269|PubMed:28719588, ECO:0007744|PDB:5MSO,
FT ECO:0007744|PDB:5MSU"
FT BINDING 960
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247,
FT ECO:0000269|PubMed:28719588, ECO:0007744|PDB:5MSO,
FT ECO:0007744|PDB:5MSU"
FT MOD_RES 685
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02247"
FT HELIX 716..725
FT /evidence="ECO:0007829|PDB:5MSU"
FT HELIX 735..739
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 748..750
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 751..753
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 756..758
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 762..767
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 768..770
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 780..784
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 789..804
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 808..813
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 815..817
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 818..828
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 834..847
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 848..852
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 858..861
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 864..873
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 876..879
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 886..888
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 890..897
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 899..908
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 910..913
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 916..921
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 922..927
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 930..932
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 935..937
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 939..942
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 944..947
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 949..951
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 954..974
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 978..983
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 985..987
FT /evidence="ECO:0007829|PDB:5MSU"
FT STRAND 990..992
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1001..1012
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 1014..1017
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 1035..1037
FT /evidence="ECO:0007829|PDB:5MSU"
FT HELIX 1038..1048
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1050..1052
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 1053..1060
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1071..1080
FT /evidence="ECO:0007829|PDB:5MSO"
FT STRAND 1086..1090
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1091..1103
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1107..1111
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1115..1121
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1136..1144
FT /evidence="ECO:0007829|PDB:5MSO"
FT TURN 1148..1151
FT /evidence="ECO:0007829|PDB:5MSO"
FT HELIX 1158..1170
FT /evidence="ECO:0007829|PDB:5MSO"
SQ SEQUENCE 1174 AA; 127797 MW; 18E2B5C6B0528AB1 CRC64;
MSPITREERL ERRIQDLYAN DPQFAAAKPA TAITAAIERP GLPLPQIIET VMTGYADRPA
LAQRSVEFVT DAGTGHTTLR LLPHFETISY GELWDRISAL ADVLSTEQTV KPGDRVCLLG
FNSVDYATID MTLARLGAVA VPLQTSAAIT QLQPIVAETQ PTMIAASVDA LADATELALS
GQTATRVLVF DHHRQVDAHR AAVESARERL AGSAVVETLA EAIARGDVPR GASAGSAPGT
DVSDDSLALL IYTSGSTGAP KGAMYPRRNV ATFWRKRTWF EGGYEPSITL NFMPMSHVMG
RQILYGTLCN GGTAYFVAKS DLSTLFEDLA LVRPTELTFV PRVWDMVFDE FQSEVDRRLV
DGADRVALEA QVKAEIRNDV LGGRYTSALT GSAPISDEMK AWVEELLDMH LVEGYGSTEA
GMILIDGAIR RPAVLDYKLV DVPDLGYFLT DRPHPRGELL VKTDSLFPGY YQRAEVTADV
FDADGFYRTG DIMAEVGPEQ FVYLDRRNNV LKLSQGEFVT VSKLEAVFGD SPLVRQIYIY
GNSARAYLLA VIVPTQEALD AVPVEELKAR LGDSLQEVAK AAGLQSYEIP RDFIIETTPW
TLENGLLTGI RKLARPQLKK HYGELLEQIY TDLAHGQADE LRSLRQSGAD APVLVTVCRA
AAALLGGSAS DVQPDAHFTD LGGDSLSALS FTNLLHEIFD IEVPVGVIVS PANDLQALAD
YVEAARKPGS SRPTFASVHG ASNGQVTEVH AGDLSLDKFI DAATLAEAPR LPAANTQVRT
VLLTGATGFL GRYLALEWLE RMDLVDGKLI CLVRAKSDTE ARARLDKTFD SGDPELLAHY
RALAGDHLEV LAGDKGEADL GLDRQTWQRL ADTVDLIVDP AALVNHVLPY SQLFGPNALG
TAELLRLALT SKIKPYSYTS TIGVADQIPP SAFTEDADIR VISATRAVDD SYANGYSNSK
WAGEVLLREA HDLCGLPVAV FRCDMILADT TWAGQLNVPD MFTRMILSLA ATGIAPGSFY
ELAADGARQR AHYDGLPVEF IAEAISTLGA QSQDGFHTYH VMNPYDDGIG LDEFVDWLNE
SGCPIQRIAD YGDWLQRFET ALRALPDRQR HSSLLPLLHN YRQPERPVRG SIAPTDRFRA
AVQEAKIGPD KDIPHVGAPI IVKYVSDLRL LGLL
//
