AKNOX_STRGJ
ID AKNOX_STRGJ Reviewed; 545 AA.
AC Q0PCD7;
DT 29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT 19-SEP-2006, sequence version 1.
DT 03-AUG-2022, entry version 75.
DE RecName: Full=Aclacinomycin-N/aclacinomycin-A oxidase {ECO:0000303|PubMed:17395717};
DE Short=AknOx {ECO:0000303|PubMed:17395717};
DE EC=1.1.3.45 {ECO:0000269|PubMed:17395717};
DE EC=1.3.3.14 {ECO:0000269|PubMed:17395717};
DE AltName: Full=Aclacinomycin oxidoreductase {ECO:0000303|PubMed:17395717};
DE Flags: Precursor;
GN Name=aknOx {ECO:0000303|PubMed:17395717};
OS Streptomyces galilaeus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=33899;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=ATCC31615;
RX PubMed=12137949; DOI=10.1016/s0378-1119(02)00699-6;
RA Raty K., Kantola J., Hautala A., Hakala J., Ylihonko K., Mantsala P.;
RT "Cloning and characterization of Streptomyces galilaeus aclacinomycins
RT polyketide synthase (PKS) cluster.";
RL Gene 293:115-122(2002).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC31615;
RA Niemi J.;
RL Submitted (AUG-2006) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP FUNCTION, DISRUPTION PHENOTYPE, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, COFACTOR, AND ACTIVITY REGULATION.
RC STRAIN=ATCC31133;
RX PubMed=528393; DOI=10.7164/antibiotics.32.472;
RA Yoshimoto A., Ogasawara T., Kitamura I., Oki T., Inui T., Takeuchi T.,
RA Umezawa H.;
RT "Enzymatic conversion of aclacinomycin A to Y by a specific oxidoreductase
RT in Streptomyces.";
RL J. Antibiot. 32:472-481(1979).
RN [4] {ECO:0007744|PDB:2IPI}
RP X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF 44-545 IN COMPLEX WITH FAD AND
RP ACLACINOMYCIN Y, PROTEIN SEQUENCE OF 44-68, FUNCTION, CATALYTIC ACTIVITY,
RP MUTAGENESIS OF TYR-187; HIS-314; GLU-417; SER-419; TYR-421 AND TYR-493,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITE, COFACTOR, SUBUNIT, AND REACTION
RP MECHANISM.
RC STRAIN=ATCC31615;
RX PubMed=17395717; DOI=10.1073/pnas.0700579104;
RA Alexeev I., Sultana A., Maentsaelae P., Niemi J., Schneider G.;
RT "Aclacinomycin oxidoreductase (AknOx) from the biosynthetic pathway of the
RT antibiotic aclacinomycin is an unusual flavoenzyme with a dual active
RT site.";
RL Proc. Natl. Acad. Sci. U.S.A. 104:6170-6175(2007).
CC -!- FUNCTION: Involved in the modification of the terminal sugar residues
CC in the last two steps in the biosynthesis of polyketide antibiotics of
CC the aclacinomycin group. In the first reaction, it catalyzes the
CC oxidation of the hydroxyl group at carbon C4 of the L-rhodinose
CC terminal sugar moiety of aclacinomycin N (AclN) to a keto group,
CC modifying the sugar to cinerulose A and generating aclacinomycin A
CC (AclA). In the second reaction, it catalyzes the elimination of two
CC hydrogen atoms from cinerulose A, leading to a double bond between
CC carbon atoms C2 and C3 and the generation of the L-aculose terminal
CC sugar moiety of aclacinomycin Y (AclY). It can also use aclacinomycin
CC analogs, epsilon-pyrromycinone glycosides, rhodirubins (A, B, C and E)
CC and all triglycosides containing L-cinerulose, L-rhodinose or 2-deoxy-
CC L-fucose as terminal sugar. {ECO:0000269|PubMed:12137949,
CC ECO:0000269|PubMed:17395717, ECO:0000269|PubMed:528393}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=aclacinomycin N + O2 = aclacinomycin A + H2O2;
CC Xref=Rhea:RHEA:37423, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:77980, ChEBI:CHEBI:77991; EC=1.1.3.45;
CC Evidence={ECO:0000269|PubMed:17395717};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=aclacinomycin A + O2 = aclacinomycin Y + H2O2;
CC Xref=Rhea:RHEA:37791, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:77980, ChEBI:CHEBI:77985; EC=1.3.3.14;
CC Evidence={ECO:0000269|PubMed:17395717};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:17395717, ECO:0000269|PubMed:528393};
CC Note=Binds 1 FAD per subunit in a bicovalent manner.
CC {ECO:0000269|PubMed:17395717};
CC -!- ACTIVITY REGULATION: Inhibited by ascorbic acid and iron ion.
CC {ECO:0000269|PubMed:528393}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=8.5 uM for AclA {ECO:0000269|PubMed:17395717};
CC Note=kcat is 0.17 sec(-1) for oxidase activity with AclA.
CC {ECO:0000269|PubMed:17395717};
CC pH dependence:
CC Optimum pH is 5.5. {ECO:0000269|PubMed:528393};
CC -!- SUBUNIT: Homotetramer; dimer of dimers. {ECO:0000269|PubMed:17395717}.
CC -!- PTM: Predicted to be exported by the Tat system. The position of the
CC signal peptide cleavage has been experimentally proven.
CC {ECO:0000255|PROSITE-ProRule:PRU00648, ECO:0000269|PubMed:17395717}.
CC -!- PTM: The FAD cofactor is bound via a bicovalent 6-S-cysteinyl, 8alpha-
CC N1-histidyl FAD linkage. {ECO:0000269|PubMed:17395717}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene accumulate various
CC aclacinomycin analogs. {ECO:0000269|PubMed:528393}.
CC -!- MISCELLANEOUS: AknOx uses two distinct sets of catalytic residues in
CC the two reactions. Tyr-493 is responsible for proton abstraction from
CC the C-4 hydroxyl group in the first reaction, the oxidation of
CC rhodinose to cinerulose A. Tyr-421 is responsible for the proton
CC abstraction from C3 of cinerulose A in the second reaction, for
CC formation L-aculose. {ECO:0000303|PubMed:17395717}.
CC -!- SIMILARITY: Belongs to the oxygen-dependent FAD-linked oxidoreductase
CC family. {ECO:0000305}.
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DR EMBL; AF257324; ABI15166.1; -; Genomic_DNA.
DR PDB; 2IPI; X-ray; 1.65 A; A/B/C/D=44-545.
DR PDBsum; 2IPI; -.
DR AlphaFoldDB; Q0PCD7; -.
DR SMR; Q0PCD7; -.
DR KEGG; ag:ABI15166; -.
DR BioCyc; MetaCyc:MON-18204; -.
DR BRENDA; 1.1.3.45; 13206.
DR BRENDA; 1.3.3.14; 13206.
DR EvolutionaryTrace; Q0PCD7; -.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 3.30.465.10; -; 1.
DR InterPro; IPR012951; BBE.
DR InterPro; IPR016166; FAD-bd_PCMH.
DR InterPro; IPR036318; FAD-bd_PCMH-like_sf.
DR InterPro; IPR016169; FAD-bd_PCMH_sub2.
DR InterPro; IPR006094; Oxid_FAD_bind_N.
DR InterPro; IPR006311; TAT_signal.
DR Pfam; PF08031; BBE; 1.
DR Pfam; PF01565; FAD_binding_4; 1.
DR SUPFAM; SSF56176; SSF56176; 1.
DR PROSITE; PS51387; FAD_PCMH; 1.
DR PROSITE; PS51318; TAT; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Direct protein sequencing; FAD;
KW Flavoprotein; Nucleotide-binding; Oxidoreductase; Signal.
FT SIGNAL 1..43
FT /note="Tat-type signal"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00648,
FT ECO:0000269|PubMed:17395717"
FT CHAIN 44..545
FT /note="Aclacinomycin-N/aclacinomycin-A oxidase"
FT /id="PRO_0000430672"
FT DOMAIN 76..256
FT /note="FAD-binding PCMH-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00718"
FT ACT_SITE 421
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:17395717"
FT ACT_SITE 493
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:17395717"
FT BINDING 451
FT /ligand="aclacinomycin Y"
FT /ligand_id="ChEBI:CHEBI:77985"
FT /evidence="ECO:0000269|PubMed:17395717,
FT ECO:0007744|PDB:2IPI"
FT BINDING 492
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17395717"
FT BINDING 493
FT /ligand="aclacinomycin Y"
FT /ligand_id="ChEBI:CHEBI:77985"
FT /evidence="ECO:0000269|PubMed:17395717,
FT ECO:0007744|PDB:2IPI"
FT CROSSLNK 113..173
FT /note="6-(S-cysteinyl)-8alpha-(pros-histidyl)-FAD (His-
FT Cys)"
FT /evidence="ECO:0000269|PubMed:17395717"
FT MUTAGEN 187
FT /note="Y->F: Loss of oxidase activity. Loss of oxidase
FT activity; when associated with F-493."
FT /evidence="ECO:0000269|PubMed:17395717"
FT MUTAGEN 314
FT /note="H->A: The oxidase activity is similar to the wild-
FT type."
FT /evidence="ECO:0000269|PubMed:17395717"
FT MUTAGEN 417
FT /note="E->A,Q: The oxidase activity is slightly decreased."
FT /evidence="ECO:0000269|PubMed:17395717"
FT MUTAGEN 419
FT /note="S->A: The oxidase activity is slightly decreased."
FT /evidence="ECO:0000269|PubMed:17395717"
FT MUTAGEN 421
FT /note="Y->F: The oxidase activity for the first reaction is
FT similar to the wild-type. Loss of oxidase activity; when
FT associated with F-493."
FT /evidence="ECO:0000269|PubMed:17395717"
FT MUTAGEN 493
FT /note="Y->F: Loss of oxidase activity. Loss of oxidase
FT activity; when associated with F-187 and F-421."
FT /evidence="ECO:0000269|PubMed:17395717"
FT STRAND 56..61
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 64..68
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 81..85
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 89..102
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 106..111
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 118..120
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 126..129
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 136..139
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 140..143
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 144..147
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 153..164
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 178..181
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 182..185
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 191..194
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 197..200
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 201..209
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 215..221
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 229..234
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 235..237
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 240..243
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 245..251
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 261..263
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 272..281
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 286..302
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 305..308
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 309..312
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 314..320
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 326..334
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 340..351
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 352..354
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 360..366
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 368..372
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 384..394
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 398..409
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 413..422
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 425..429
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 432..434
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 443..453
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 455..457
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 458..472
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 473..476
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 477..479
FT /evidence="ECO:0007829|PDB:2IPI"
FT STRAND 482..486
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 496..499
FT /evidence="ECO:0007829|PDB:2IPI"
FT TURN 501..503
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 510..515
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 516..518
FT /evidence="ECO:0007829|PDB:2IPI"
FT HELIX 519..529
FT /evidence="ECO:0007829|PDB:2IPI"
SQ SEQUENCE 545 AA; 59014 MW; A51AE3C6E6229EFF CRC64;
MFVLNEFTRR GFLGTAAAVG GTTVVTTALG GAPAAQAAVP EAADGGACGA RTALVKVDRV
DRRYQDLVTR GFNGRFRGRP DVVYVVHTAD QVVDAVNQAM AAGQRIAVRS GGHCFEGFVD
DPAVRAVIDM SQMRQVFYDS GKRAFAVEPG ATLGETYRAL YLDWGVTIPA GVCPQVGVGG
HVLGGGYGPL SRRDGVVADH LYAVEVVVVD ASGRARKVVA TSAADDPNRE LWWAHTGGGG
GNFGIVTRYW FRTPGATGTD PSQLLPKAPT STLRHIVTWD WSALTEEAFT RIIDNHGAWH
QSNSAAGTPY ASMHSVFYLN SRAAGQILLD IQIDGGLDGA EALLNDFVAA VNEGTGVEPA
VQRSTEPWLR ATLANKFDTG GFDRTKSKGA YLRKPWTAAQ AATLYRHLSA DSQVWGEVSL
YSYGGKVNSV PETATATAQR DSIIKVWMSA TWMDPAHDDA NLAWIREIYR EIFATTGGVP
VPDDRTEGTF INYPDVDLVD ERWNTSGVPW YTLYYKGNYP RLQKVKARWD PRDVFRHALS
VRPPG
