ROSB_STRDJ
ID ROSB_STRDJ Reviewed; 257 AA.
AC K4REZ6;
DT 20-JUN-2018, integrated into UniProtKB/Swiss-Prot.
DT 09-JAN-2013, sequence version 1.
DT 03-AUG-2022, entry version 54.
DE RecName: Full=8-demethyl-8-aminoriboflavin-5'-phosphate synthase {ECO:0000303|PubMed:27062037};
DE Short=AFP synthase {ECO:0000303|PubMed:27062037};
DE EC=2.6.1.114 {ECO:0000269|PubMed:27062037, ECO:0000269|PubMed:27981706, ECO:0000305|PubMed:27331868};
DE AltName: Full=8-amino-flavin synthase {ECO:0000303|PubMed:27331868};
GN Name=rosB {ECO:0000303|PubMed:27062037};
GN Synonyms=Orf7989 {ECO:0000303|PubMed:27331868};
GN ORFNames=BN159_7989 {ECO:0000312|EMBL:CCK32368.1};
OS Streptomyces davaonensis (strain DSM 101723 / JCM 4913 / KCC S-0913 / 768).
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1214101;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 101723 / JCM 4913 / KCC S-0913 / 768;
RX PubMed=23043000; DOI=10.1128/jb.01592-12;
RA Jankowitsch F., Schwarz J., Ruckert C., Gust B., Szczepanowski R., Blom J.,
RA Pelzer S., Kalinowski J., Mack M.;
RT "Genome sequence of the bacterium Streptomyces davawensis JCM 4913 and
RT heterologous production of the unique antibiotic roseoflavin.";
RL J. Bacteriol. 194:6818-6827(2012).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, DISRUPTION PHENOTYPE, PATHWAY, SUBUNIT, AND
RP REACTION MECHANISM.
RC STRAIN=DSM 101723 / JCM 4913 / KCC S-0913 / 768;
RX PubMed=27062037; DOI=10.1002/anie.201600581;
RA Schwarz J., Konjik V., Jankowitsch F., Sandhoff R., Mack M.;
RT "Identification of the Key enzyme of roseoflavin biosynthesis.";
RL Angew. Chem. Int. Ed. 55:6103-6106(2016).
RN [3]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=DSM 101723 / JCM 4913 / KCC S-0913 / 768;
RX PubMed=27331868; DOI=10.1021/jacs.6b02469;
RA Jhulki I., Chanani P.K., Abdelwahed S.H., Begley T.P.;
RT "A remarkable oxidative cascade that replaces the riboflavin C8 methyl with
RT an amino group during roseoflavin biosynthesis.";
RL J. Am. Chem. Soc. 138:8324-8327(2016).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG,
RP FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF THR-11; ARG-13; SER-19;
RP THR-21; ASN-94 AND TYR-240, SUBUNIT, AND REACTION MECHANISM.
RC STRAIN=DSM 101723 / JCM 4913 / KCC S-0913 / 768;
RX PubMed=27981706; DOI=10.1002/anie.201610292;
RA Konjik V., Brunle S., Demmer U., Vanselow A., Sandhoff R., Ermler U.,
RA Mack M.;
RT "The crystal structure of RosB: insights into the reaction mechanism of the
RT first member of a family of flavodoxin-like enzymes.";
RL Angew. Chem. Int. Ed. 56:1146-1151(2017).
CC -!- FUNCTION: Involved in the biosynthesis of the riboflavin analog
CC antibiotic roseoflavin (3,8-dimethylamino-riboflavin)
CC (PubMed:27062037). Catalyzes the site-specific substitution of the C-8
CC methyl group of riboflavin-5'-phosphate (FMN) by an amino group to
CC yield 8-amino-8-demethylriboflavin 5'-phosphate, via a combined
CC oxidation, decarboxylation and transamination reaction
CC (PubMed:27062037, PubMed:27331868, PubMed:27981706). The catalysis is
CC initiated by an oxidation step in which the C-8 methyl group on the
CC dimethylbenzene ring of FMN is converted to a formyl group to yield the
CC 8-demethyl-8-formylriboflavin-5'-phosphate (OHC-RP) intermediate
CC (PubMed:27062037). In the presence of thiamine, the formyl group is
CC oxidized into a carboxyl group to yield the 8-demethyl-8-
CC carboxyriboflavin-5'-phosphate (HO2C-RP) intermediate
CC (PubMed:27062037). Finally, in the presence of L-glutamate as an amino
CC donor, decarboxylation and aminotransfer occur, resulting in production
CC of 8-demethyl-8-aminoriboflavin-5'-phosphate (PubMed:27062037).
CC Addition of NAD (but not NADP) to the reaction increases the yield 1.7-
CC fold (PubMed:27062037). The reaction also proceeds without the addition
CC of any electron acceptor, and it is possible that molecular oxygen
CC serves this role (PubMed:27062037). {ECO:0000269|PubMed:27062037,
CC ECO:0000269|PubMed:27331868, ECO:0000269|PubMed:27981706}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3 A + FMN + H2O + L-glutamate + O2 = 2-oxoglutarate + 8-amino-
CC 8-demethylriboflavin 5'-phosphate + 3 AH2 + CO2 + H(+);
CC Xref=Rhea:RHEA:54992, ChEBI:CHEBI:13193, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16526,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:17499, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:58210, ChEBI:CHEBI:139569; EC=2.6.1.114;
CC Evidence={ECO:0000269|PubMed:27062037, ECO:0000269|PubMed:27981706,
CC ECO:0000305|PubMed:27331868};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000269|PubMed:27062037}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:27062037,
CC ECO:0000269|PubMed:27981706}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene are unable to produce 8-
CC demethyl-8-aminoriboflavin (AF). {ECO:0000269|PubMed:27062037}.
CC -!- SIMILARITY: Belongs to the SsuE family. {ECO:0000305}.
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DR EMBL; HE971709; CCK32368.1; -; Genomic_DNA.
DR RefSeq; WP_015662694.1; NC_020504.1.
DR PDB; 5MJI; X-ray; 2.00 A; A=1-257.
DR PDB; 5MLD; X-ray; 1.70 A; A/B/C/D/E/F/G/H=1-257.
DR PDBsum; 5MJI; -.
DR PDBsum; 5MLD; -.
DR AlphaFoldDB; K4REZ6; -.
DR SMR; K4REZ6; -.
DR STRING; 1214101.BN159_7989; -.
DR EnsemblBacteria; CCK32368; CCK32368; BN159_7989.
DR KEGG; sdv:BN159_7989; -.
DR PATRIC; fig|1214101.3.peg.8085; -.
DR eggNOG; COG0655; Bacteria.
DR HOGENOM; CLU_082329_0_0_11; -.
DR OMA; CVANTFA; -.
DR OrthoDB; 796211at2; -.
DR BioCyc; MetaCyc:BN159_RS39585-MON; -.
DR BRENDA; 2.6.1.114; 9909.
DR Proteomes; UP000008043; Chromosome.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:InterPro.
DR GO; GO:0008483; F:transaminase activity; IDA:UniProtKB.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0016999; P:antibiotic metabolic process; IDA:UniProtKB.
DR Gene3D; 3.40.50.360; -; 1.
DR InterPro; IPR029039; Flavoprotein-like_sf.
DR InterPro; IPR005025; FMN_Rdtase-like.
DR Pfam; PF03358; FMN_red; 1.
DR SUPFAM; SSF52218; SSF52218; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Flavoprotein; FMN;
KW Reference proteome; Transferase.
FT CHAIN 1..257
FT /note="8-demethyl-8-aminoriboflavin-5'-phosphate synthase"
FT /id="PRO_0000444475"
FT BINDING 11..13
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:27981706,
FT ECO:0007744|PDB:5MJI, ECO:0007744|PDB:5MLD"
FT BINDING 19..21
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:27981706,
FT ECO:0007744|PDB:5MJI, ECO:0007744|PDB:5MLD"
FT BINDING 91..94
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:27981706,
FT ECO:0007744|PDB:5MJI, ECO:0007744|PDB:5MLD"
FT BINDING 132..136
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:27981706,
FT ECO:0007744|PDB:5MJI, ECO:0007744|PDB:5MLD"
FT BINDING 240
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:27981706"
FT SITE 13
FT /note="Important to position the amino-group donor
FT glutamate"
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 11
FT /note="T->A: Loss of synthase activity due to the absence
FT of substrate binding."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 13
FT /note="R->A: Loss of synthase activity due to the reduced
FT ability to catalyze the introduction of an amino group and
FT predominantly to synthesize 8-demethyl-8-carboxyriboflavin-
FT 5'-phosphate (HO2C-RP) intermediate."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 19
FT /note="S->A: Loss of synthase activity due to the absence
FT of substrate binding."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 21
FT /note="T->A: Strong decrease of synthase activity."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 53
FT /note="Y->A: Loss of synthase activity."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 55
FT /note="D->A: Loss of synthase activity due to the absence
FT of substrate binding."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 94
FT /note="N->A: Loss of synthase activity due to the absence
FT of substrate binding."
FT /evidence="ECO:0000269|PubMed:27981706"
FT MUTAGEN 240
FT /note="Y->A: Loss of synthase activity due to the absence
FT of substrate binding."
FT /evidence="ECO:0000269|PubMed:27981706"
FT STRAND 4..9
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 20..34
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 38..43
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 44..46
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 62..64
FT /evidence="ECO:0007829|PDB:5MLD"
FT TURN 65..67
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 72..81
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 83..92
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 98..107
FT /evidence="ECO:0007829|PDB:5MLD"
FT TURN 115..117
FT /evidence="ECO:0007829|PDB:5MLD"
FT TURN 121..124
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 125..132
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 134..136
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 138..152
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 154..156
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 161..164
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 167..170
FT /evidence="ECO:0007829|PDB:5MLD"
FT TURN 174..178
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 179..181
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 183..205
FT /evidence="ECO:0007829|PDB:5MLD"
FT HELIX 212..220
FT /evidence="ECO:0007829|PDB:5MLD"
FT STRAND 223..230
FT /evidence="ECO:0007829|PDB:5MLD"
SQ SEQUENCE 257 AA; 28865 MW; F92673D1CC9EC343 CRC64;
MALKALILNT TLRRSPSRSQ TQGLIDKAVP LYEKEGIETE VVRVIDHDIE QEYWDDYDDW
NAGEKARRED EWPWLLEKIR EADILVIATP ITLNMCTSAA HVILEKLNLM DELNGDTKQF
PLYNKVAGLL MCGNEDGAHH VAGTVLNNLG RLGYSVPPNA AAYWLGPAGT GPGYIEGKGD
RHFHTNKLIR FMVANTSHLA RMLQETPYTT DLEACAQAAR EESDDVFAIR VNVNTPAIRY
KRFQKLGEVK VEESQLG
