BLH_UNCMB
ID BLH_UNCMB Reviewed; 275 AA.
AC Q4PNI0;
DT 31-MAY-2011, integrated into UniProtKB/Swiss-Prot.
DT 19-JUL-2005, sequence version 1.
DT 03-AUG-2022, entry version 36.
DE RecName: Full=Beta-carotene 15,15'-dioxygenase {ECO:0000305};
DE EC=1.13.11.63 {ECO:0000255|HAMAP-Rule:MF_02093, ECO:0000269|PubMed:19366683};
GN Name=blh;
OS Uncultured marine bacterium 66A03.
OC Bacteria; environmental samples.
OX NCBI_TaxID=331677;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=16008504; DOI=10.1371/journal.pbio.0030273;
RA Sabehi G., Loy A., Jung K.H., Partha R., Spudich J.L., Isaacson T.,
RA Hirschberg J., Wagner M., Beja O.;
RT "New insights into metabolic properties of marine bacteria encoding
RT proteorhodopsins.";
RL PLoS Biol. 3:E273-E273(2005).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, COFACTOR,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, METAL-BINDING SITES, REACTION
RP MECHANISM, AND MUTAGENESIS OF HIS-21; HIS-78; HIS-188 AND HIS-192.
RX PubMed=19366683; DOI=10.1074/jbc.m109.002618;
RA Kim Y.S., Kim N.H., Yeom S.J., Kim S.W., Oh D.K.;
RT "In vitro characterization of a recombinant Blh protein from an uncultured
RT marine bacterium as a beta-carotene 15,15'-dioxygenase.";
RL J. Biol. Chem. 284:15781-15793(2009).
RN [3]
RP BIOTECHNOLOGY.
RX PubMed=20229064; DOI=10.1007/s10529-010-0239-3;
RA Kim Y.S., Park C.S., Oh D.K.;
RT "Retinal production from beta-carotene by beta-carotene 15,15'-dioxygenase
RT from an unculturable marine bacterium.";
RL Biotechnol. Lett. 32:957-961(2010).
CC -!- FUNCTION: Catalyzes the cleavage of beta-carotene at its central double
CC bond (15,15') to yield two molecules of all-trans-retinal. Exhibits the
CC highest activity for beta-carotene, followed by beta-cryptoxanthin,
CC beta-apo-4'-carotenal, alpha-carotene, and gamma-carotene in decreasing
CC order, but has no activity on beta-apo-8'-carotenal, beta-apo-12'-
CC carotenal, lutein, zeaxanthin, or lycopene, suggesting that the
CC presence of one unsubstituted beta-ionone ring in a substrate with a
CC molecular weight greater than C35 seems to be essential for enzyme
CC activity. {ECO:0000269|PubMed:19366683}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=all-trans-beta-carotene + O2 = 2 all-trans-retinal;
CC Xref=Rhea:RHEA:32887, ChEBI:CHEBI:15379, ChEBI:CHEBI:17579,
CC ChEBI:CHEBI:17898; EC=1.13.11.63; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_02093, ECO:0000269|PubMed:19366683};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_02093,
CC ECO:0000269|PubMed:19366683};
CC Note=Binds 1 Fe(2+) ion per subunit. {ECO:0000269|PubMed:19366683};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=37 uM for beta-carotene {ECO:0000269|PubMed:19366683};
CC KM=29 uM for beta-cryptoxanthin {ECO:0000269|PubMed:19366683};
CC KM=147 uM for beta-apo-4'-carotenal {ECO:0000269|PubMed:19366683};
CC KM=169 uM for alpha-carotene {ECO:0000269|PubMed:19366683};
CC KM=382 uM for gamma-carotene {ECO:0000269|PubMed:19366683};
CC Note=The kcat is 4.5-fold higher with beta-carotene than with beta-
CC cryptoxanthin as substrate.;
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:19366683};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius. Above this temperature,
CC the enzyme activity decreases significantly, exhibiting 66% of the
CC maximum activity at 50 degrees Celsius. Below 40 degrees Celsius, the
CC enzyme activity decreases with decreasing temperature, exhibiting 15%
CC of the maximum activity at 25 degrees Celsius. The half-lives of the
CC enzyme at 35, 40, 45, 50, and 55 degrees Celsius are 17.6, 15.0,
CC 12.5, 6.1, and 1.5 hours, respectively.
CC {ECO:0000269|PubMed:19366683};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:19366683}.
CC -!- SUBCELLULAR LOCATION: Cell membrane {ECO:0000255|HAMAP-Rule:MF_02093,
CC ECO:0000305}; Multi-pass membrane protein {ECO:0000255|HAMAP-
CC Rule:MF_02093, ECO:0000305}.
CC -!- BIOTECHNOLOGY: Could be used for high retinal production.
CC {ECO:0000269|PubMed:20229064}.
CC -!- SIMILARITY: Belongs to the Brp/Blh beta-carotene diooxygenase family.
CC {ECO:0000255|HAMAP-Rule:MF_02093, ECO:0000305}.
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DR EMBL; DQ065755; AAY68319.1; -; Genomic_DNA.
DR AlphaFoldDB; Q4PNI0; -.
DR KEGG; ag:AAY68319; -.
DR BioCyc; MetaCyc:MON-17377; -.
DR BRENDA; 1.13.11.63; 3186.
DR GO; GO:0005887; C:integral component of plasma membrane; IEA:UniProtKB-UniRule.
DR GO; GO:0003834; F:beta-carotene 15,15'-dioxygenase activity; IEA:UniProtKB-EC.
DR GO; GO:0010436; F:carotenoid dioxygenase activity; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0016121; P:carotene catabolic process; IDA:UniProtKB.
DR GO; GO:0042574; P:retinal metabolic process; IDA:UniProtKB.
DR HAMAP; MF_02093; Beta_carotene_diox; 1.
DR InterPro; IPR022270; Blh_monoox.
DR Pfam; PF15461; BCD; 1.
DR TIGRFAMs; TIGR03753; blh_monoox; 1.
PE 1: Evidence at protein level;
KW Cell membrane; Dioxygenase; Iron; Membrane; Metal-binding; Oxidoreductase;
KW Transmembrane; Transmembrane helix.
FT CHAIN 1..275
FT /note="Beta-carotene 15,15'-dioxygenase"
FT /id="PRO_0000408499"
FT TRANSMEM 1..21
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 36..56
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 57..77
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 91..111
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 127..147
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 160..180
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 210..230
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT TRANSMEM 244..264
FT /note="Helical"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093"
FT BINDING 21
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093,
FT ECO:0000305|PubMed:19366683"
FT BINDING 78
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093,
FT ECO:0000305|PubMed:19366683"
FT BINDING 188
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093,
FT ECO:0000305|PubMed:19366683"
FT BINDING 192
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_02093,
FT ECO:0000305|PubMed:19366683"
FT MUTAGEN 21
FT /note="H->A: 3% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:19366683"
FT MUTAGEN 78
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:19366683"
FT MUTAGEN 188
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:19366683"
FT MUTAGEN 192
FT /note="H->A: 5% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:19366683"
SQ SEQUENCE 275 AA; 31037 MW; D571B00FCA434036 CRC64;
MGLMLIDWCA LALVVFIGLP HGALDAAISF SMISSAKRIA RLAGILLIYL LLATAFFLIW
YQLPAFSLLI FLLISIIHFG MADFNASPSK LKWPHIIAHG GVVTVWLPLI QKNEVTKLFS
ILTNGPTPIL WDILLIFFLC WSIGVCLHTY ETLRSKHYNI AFELIGLIFL AWYAPPLVTF
ATYFCFIHSR RHFSFVWKQL QHMSSKKMMI GSAIILSCTS WLIGGGIYFF LNSKMIASEA
ALQTVFIGLA ALTVPHMILI DFIFRPHSSR IKIKN
