PHAB_CUPNH
ID PHAB_CUPNH Reviewed; 246 AA.
AC P14697; Q0KBP7;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-APR-1990, sequence version 1.
DT 03-AUG-2022, entry version 148.
DE RecName: Full=Acetoacetyl-CoA reductase;
DE EC=1.1.1.36;
GN Name=phaB {ECO:0000303|PubMed:1476773};
GN Synonyms=phbB {ECO:0000303|PubMed:2670935}; OrderedLocusNames=H16_A1439;
OS Cupriavidus necator (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442
OS / H16 / Stanier 337) (Ralstonia eutropha).
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Burkholderiaceae; Cupriavidus.
OX NCBI_TaxID=381666;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION, AND FUNCTION.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=2670935; DOI=10.1016/s0021-9258(19)84824-x;
RA Peoples O.P., Sinskey A.J.;
RT "Poly-beta-hydroxybutyrate biosynthesis in Alcaligenes eutrophus H16.
RT Characterization of the genes encoding beta-ketothiolase and acetoacetyl-
RT CoA reductase.";
RL J. Biol. Chem. 264:15293-15297(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=16964242; DOI=10.1038/nbt1244;
RA Pohlmann A., Fricke W.F., Reinecke F., Kusian B., Liesegang H., Cramm R.,
RA Eitinger T., Ewering C., Poetter M., Schwartz E., Strittmatter A., Voss I.,
RA Gottschalk G., Steinbuechel A., Friedrich B., Bowien B.;
RT "Genome sequence of the bioplastic-producing 'Knallgas' bacterium Ralstonia
RT eutropha H16.";
RL Nat. Biotechnol. 24:1257-1262(2006).
RN [3]
RP FUNCTION IN PHB SYNTHESIS, AND PATHWAY.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=2670936; DOI=10.1016/s0021-9258(19)84825-1;
RA Peoples O.P., Sinskey A.J.;
RT "Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16.
RT Identification and characterization of the PHB polymerase gene (phbC).";
RL J. Biol. Chem. 264:15298-15303(1989).
RN [4]
RP GENE NAME.
RX PubMed=1476773; DOI=10.1111/j.1574-6968.1992.tb05841.x;
RA Steinbuechel A., Hustede E., Liebergesell M., Pieper U., Timm A.,
RA Valentin H.;
RT "Molecular basis for biosynthesis and accumulation of polyhydroxyalkanoic
RT acids in bacteria.";
RL FEMS Microbiol. Rev. 9:217-230(1992).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.79 ANGSTROMS) OF WILD-TYPE AND MUTANTS LEU-47 AND
RP SER-173 IN COMPLEX WITH ACETOACETYL-COENZYME A AND NADP, FUNCTION,
RP CATALYTIC ACTIVITY, KINETIC PARAMETERS, SUBUNIT, PATHWAY, AND MUTAGENESIS
RP OF GLN-47 AND THR-173.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=23913421; DOI=10.1128/aem.01768-13;
RA Matsumoto K., Tanaka Y., Watanabe T., Motohashi R., Ikeda K., Tobitani K.,
RA Yao M., Tanaka I., Taguchi S.;
RT "Directed evolution and structural analysis of NADPH-dependent acetoacetyl
RT coenzyme A (acetoacetyl-CoA) reductase from Ralstonia eutropha reveals two
RT mutations responsible for enhanced kinetics.";
RL Appl. Environ. Microbiol. 79:6134-6139(2013).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.34 ANGSTROMS) OF APOENZYME AND IN COMPLEXES WITH
RP ACETOACETYL-COENZYME A AND NADP, SUBUNIT, FUNCTION, CATALYTIC ACTIVITY, AND
RP MUTAGENESIS OF GLN-47; ASP-94; LYS-99; GLN-147; PHE-148; GLN-150; THR-173;
RP TYR-185 AND ARG-195.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=24211201; DOI=10.1016/j.bbrc.2013.10.150;
RA Kim J., Chang J.H., Kim E.J., Kim K.J.;
RT "Crystal structure of (R)-3-hydroxybutyryl-CoA dehydrogenase PhaB from
RT Ralstonia eutropha.";
RL Biochem. Biophys. Res. Commun. 443:783-788(2014).
CC -!- FUNCTION: Catalyzes the chiral reduction of acetoacetyl-CoA to (R)-3-
CC hydroxybutyryl-CoA. Is involved in the biosynthesis of
CC polyhydroxybutyrate (PHB), which is accumulated as an intracellular
CC energy reserve material when cells grow under conditions of nutrient
CC limitation. {ECO:0000269|PubMed:23913421, ECO:0000269|PubMed:24211201,
CC ECO:0000269|PubMed:2670935, ECO:0000269|PubMed:2670936}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a (3R)-3-hydroxyacyl-CoA + NADP(+) = a 3-oxoacyl-CoA + H(+) +
CC NADPH; Xref=Rhea:RHEA:22256, ChEBI:CHEBI:15378, ChEBI:CHEBI:57319,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:90726; EC=1.1.1.36;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(3R)-3-hydroxybutanoyl-CoA + NADP(+) = acetoacetyl-CoA + H(+)
CC + NADPH; Xref=Rhea:RHEA:45796, ChEBI:CHEBI:15378, ChEBI:CHEBI:57286,
CC ChEBI:CHEBI:57315, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.1.1.36;
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.7 uM for acetoacetyl-CoA {ECO:0000269|PubMed:23913421};
CC KM=149 uM for NADPH {ECO:0000269|PubMed:23913421};
CC Note=kcat is 102 sec(-1).;
CC -!- PATHWAY: Biopolymer metabolism; poly-(R)-3-hydroxybutanoate
CC biosynthesis. {ECO:0000269|PubMed:23913421,
CC ECO:0000269|PubMed:2670936}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:23913421,
CC ECO:0000269|PubMed:24211201}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; J04987; AAA21973.1; -; Genomic_DNA.
DR EMBL; AM260479; CAJ92574.1; -; Genomic_DNA.
DR PIR; B34340; RDALAE.
DR RefSeq; WP_010810131.1; NZ_CP039287.1.
DR PDB; 3VZP; X-ray; 1.79 A; A/B/C/D=2-246.
DR PDB; 3VZQ; X-ray; 2.00 A; A/B=2-246.
DR PDB; 3VZR; X-ray; 2.90 A; A/B=2-246.
DR PDB; 3VZS; X-ray; 2.14 A; A/B/C/D=2-246.
DR PDB; 4N5L; X-ray; 1.65 A; A/B=1-246.
DR PDB; 4N5M; X-ray; 1.34 A; A/B=1-246.
DR PDB; 4N5N; X-ray; 1.90 A; A/B=1-246.
DR PDBsum; 3VZP; -.
DR PDBsum; 3VZQ; -.
DR PDBsum; 3VZR; -.
DR PDBsum; 3VZS; -.
DR PDBsum; 4N5L; -.
DR PDBsum; 4N5M; -.
DR PDBsum; 4N5N; -.
DR AlphaFoldDB; P14697; -.
DR SMR; P14697; -.
DR STRING; 381666.H16_A1439; -.
DR PRIDE; P14697; -.
DR EnsemblBacteria; CAJ92574; CAJ92574; H16_A1439.
DR GeneID; 57643538; -.
DR KEGG; reh:H16_A1439; -.
DR eggNOG; COG1028; Bacteria.
DR HOGENOM; CLU_010194_1_3_4; -.
DR OMA; EFHSCDV; -.
DR OrthoDB; 1601931at2; -.
DR BRENDA; 1.1.1.100; 231.
DR BRENDA; 1.1.1.36; 231.
DR UniPathway; UPA00917; -.
DR Proteomes; UP000008210; Chromosome 1.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0018454; F:acetoacetyl-CoA reductase activity; IEA:UniProtKB-EC.
DR GO; GO:0042619; P:poly-hydroxybutyrate biosynthetic process; IEA:UniProtKB-KW.
DR InterPro; IPR011283; Acetoacetyl-CoA_reductase.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR020904; Sc_DH/Rdtase_CS.
DR InterPro; IPR002347; SDR_fam.
DR Pfam; PF00106; adh_short; 1.
DR PRINTS; PR00081; GDHRDH.
DR PRINTS; PR00080; SDRFAMILY.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR01829; AcAcCoA_reduct; 1.
DR PROSITE; PS00061; ADH_SHORT; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; NADP; Oxidoreductase; PHB biosynthesis;
KW Reference proteome.
FT CHAIN 1..246
FT /note="Acetoacetyl-CoA reductase"
FT /id="PRO_0000054748"
FT ACT_SITE 153
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10001"
FT BINDING 13..15
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23913421"
FT BINDING 35
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23913421"
FT BINDING 40
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23913421"
FT BINDING 60..62
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23913421"
FT BINDING 88..92
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23913421"
FT BINDING 94
FT /ligand="substrate"
FT BINDING 147..150
FT /ligand="substrate"
FT BINDING 183..186
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23913421"
FT BINDING 184..185
FT /ligand="substrate"
FT BINDING 195
FT /ligand="substrate"
FT MUTAGEN 47
FT /note="Q->L: 2.4-fold increase in activity. 2-fold decrease
FT in affinity for NADPH and 2.8-fold decrease in affinity for
FT acetoacetyl-CoA."
FT /evidence="ECO:0000269|PubMed:23913421,
FT ECO:0000269|PubMed:24211201"
FT MUTAGEN 94
FT /note="D->A: About 6% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT MUTAGEN 99
FT /note="K->A: Nearly loss of activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT MUTAGEN 147
FT /note="Q->A: About 30% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT MUTAGEN 148
FT /note="F->A: About 30% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT MUTAGEN 150
FT /note="Q->A: About 20% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT MUTAGEN 173
FT /note="T->S: 3.5-fold increase in activity. 4-fold decrease
FT in affinity for NADPH and 2.4-fold decrease in affinity for
FT acetoacetyl-CoA."
FT /evidence="ECO:0000269|PubMed:23913421,
FT ECO:0000269|PubMed:24211201"
FT MUTAGEN 185
FT /note="Y->A: Nearly loss of activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT MUTAGEN 195
FT /note="R->A: Nearly loss of activity."
FT /evidence="ECO:0000269|PubMed:24211201"
FT STRAND 5..8
FT /evidence="ECO:0007829|PDB:4N5M"
FT TURN 9..12
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 14..25
FT /evidence="ECO:0007829|PDB:4N5M"
FT STRAND 29..34
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 41..50
FT /evidence="ECO:0007829|PDB:4N5M"
FT STRAND 56..59
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 65..78
FT /evidence="ECO:0007829|PDB:4N5M"
FT STRAND 82..87
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 97..99
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 102..112
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 114..130
FT /evidence="ECO:0007829|PDB:4N5M"
FT STRAND 133..138
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 141..145
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 151..171
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 172..174
FT /evidence="ECO:0007829|PDB:4N5M"
FT STRAND 176..183
FT /evidence="ECO:0007829|PDB:4N5M"
FT STRAND 185..188
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 189..192
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 196..204
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 214..225
FT /evidence="ECO:0007829|PDB:4N5M"
FT HELIX 227..229
FT /evidence="ECO:0007829|PDB:3VZP"
FT STRAND 236..240
FT /evidence="ECO:0007829|PDB:4N5M"
SQ SEQUENCE 246 AA; 26370 MW; AD6739E0F5C93C06 CRC64;
MTQRIAYVTG GMGGIGTAIC QRLAKDGFRV VAGCGPNSPR REKWLEQQKA LGFDFIASEG
NVADWDSTKT AFDKVKSEVG EVDVLINNAG ITRDVVFRKM TRADWDAVID TNLTSLFNVT
KQVIDGMADR GWGRIVNISS VNGQKGQFGQ TNYSTAKAGL HGFTMALAQE VATKGVTVNT
VSPGYIATDM VKAIRQDVLD KIVATIPVKR LGLPEEIASI CAWLSSEESG FSTGADFSLN
GGLHMG
