LADA_GEOTN
ID LADA_GEOTN Reviewed; 440 AA.
AC A4IU28;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2007, sequence version 1.
DT 03-AUG-2022, entry version 92.
DE RecName: Full=Long-chain alkane monooxygenase {ECO:0000303|PubMed:17372208};
DE EC=1.14.14.28 {ECO:0000269|PubMed:17372208, ECO:0000269|PubMed:18164311, ECO:0000269|PubMed:22526792};
DE AltName: Full=Long-chain alkane degradation protein A {ECO:0000305};
GN Name=ladA {ECO:0000303|PubMed:17372208};
GN OrderedLocusNames=GTNG_3499 {ECO:0000312|EMBL:ABO68832.1};
OS Geobacillus thermodenitrificans (strain NG80-2).
OG Plasmid pLW1071 {ECO:0000312|EMBL:ABO68832.1}.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Geobacillus.
OX NCBI_TaxID=420246;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA], FUNCTION, CATALYTIC
RP ACTIVITY, SUBCELLULAR LOCATION, INDUCTION, AND BIOTECHNOLOGY.
RC STRAIN=NG80-2;
RX PubMed=17372208; DOI=10.1073/pnas.0609650104;
RA Feng L., Wang W., Cheng J., Ren Y., Zhao G., Gao C., Tang Y., Liu X.,
RA Han W., Peng X., Liu R., Wang L.;
RT "Genome and proteome of long-chain alkane degrading Geobacillus
RT thermodenitrificans NG80-2 isolated from a deep-subsurface oil reservoir.";
RL Proc. Natl. Acad. Sci. U.S.A. 104:5602-5607(2007).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY,
RP AND MUTAGENESIS OF ALA-102; PHE-146; LEU-320 AND ASN-376.
RC STRAIN=NG80-2;
RX PubMed=22526792; DOI=10.1007/s00253-012-4035-y;
RA Dong Y., Yan J., Du H., Chen M., Ma T., Feng L.;
RT "Engineering of LadA for enhanced hexadecane oxidation using random- and
RT site-directed mutagenesis.";
RL Appl. Microbiol. Biotechnol. 94:1019-1029(2012).
RN [3] {ECO:0007744|PDB:3B9N, ECO:0007744|PDB:3B9O}
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH
RP FMN, CATALYTIC ACTIVITY, SUBUNIT, AND MUTAGENESIS OF CYS-14; HIS-17;
RP TYR-63; GLN-79 AND HIS-311.
RX PubMed=18164311; DOI=10.1016/j.jmb.2007.11.069;
RA Li L., Liu X., Yang W., Xu F., Wang W., Feng L., Bartlam M., Wang L.,
RA Rao Z.;
RT "Crystal structure of long-chain alkane monooxygenase (LadA) in complex
RT with coenzyme FMN: unveiling the long-chain alkane hydroxylase.";
RL J. Mol. Biol. 376:453-465(2008).
CC -!- FUNCTION: Involved in the degradation of long-chain alkanes
CC (PubMed:17372208, PubMed:22526792). Converts alkanes ranging from C(15)
CC to C(36) into their corresponding primary alcohols (PubMed:17372208).
CC {ECO:0000269|PubMed:17372208, ECO:0000269|PubMed:22526792}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a long-chain alkane + FMNH2 + O2 = a long chain fatty alcohol
CC + FMN + H(+) + H2O; Xref=Rhea:RHEA:49060, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:17135,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:83563;
CC EC=1.14.14.28; Evidence={ECO:0000269|PubMed:17372208,
CC ECO:0000269|PubMed:18164311, ECO:0000269|PubMed:22526792};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49061;
CC Evidence={ECO:0000269|PubMed:17372208, ECO:0000269|PubMed:18164311,
CC ECO:0000269|PubMed:22526792};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=9.1 mM for hexadecane {ECO:0000269|PubMed:22526792};
CC Note=kcat is 1.3 min(-1) with hexadecane as substrate.
CC {ECO:0000269|PubMed:22526792};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:22526792};
CC Temperature dependence:
CC Optimum temperature is 60 degrees Celsius.
CC {ECO:0000269|PubMed:22526792};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:18164311}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:17372208}.
CC -!- INDUCTION: 120-fold increase in transcription when crude oil is used as
CC a sole carbon source instead of sucrose. {ECO:0000269|PubMed:17372208}.
CC -!- BIOTECHNOLOGY: Shows particular potential to be used for the treatment
CC of environmental oil pollutions and in other biocatalytic processes
CC (PubMed:17372208, PubMed:22526792). Random- and site-directed
CC mutagenesis were used to generate mutants with increased activity
CC toward hexadecane (PubMed:22526792). {ECO:0000269|PubMed:17372208,
CC ECO:0000269|PubMed:22526792}.
CC -!- SIMILARITY: Belongs to the NtaA/SnaA/DszA monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; CP000558; ABO68832.1; -; Genomic_DNA.
DR RefSeq; WP_011888513.1; NC_009329.1.
DR PDB; 3B9N; X-ray; 2.70 A; A/B=1-440.
DR PDB; 3B9O; X-ray; 1.90 A; A/B=1-440.
DR PDBsum; 3B9N; -.
DR PDBsum; 3B9O; -.
DR SMR; A4IU28; -.
DR STRING; 420246.GTNG_3499; -.
DR EnsemblBacteria; ABO68832; ABO68832; GTNG_3499.
DR KEGG; gtn:GTNG_3499; -.
DR eggNOG; COG2141; Bacteria.
DR HOGENOM; CLU_022256_0_0_9; -.
DR OMA; RVETNAI; -.
DR OrthoDB; 1653347at2; -.
DR BRENDA; 1.14.14.28; 705.
DR EvolutionaryTrace; A4IU28; -.
DR Proteomes; UP000001578; Plasmid pLW1071.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro.
DR CDD; cd01095; Nitrilotriacetate_monoxgenase; 1.
DR Gene3D; 3.20.20.30; -; 1.
DR InterPro; IPR011251; Luciferase-like_dom.
DR InterPro; IPR036661; Luciferase-like_sf.
DR InterPro; IPR016215; NTA_MOA.
DR Pfam; PF00296; Bac_luciferase; 1.
DR PIRSF; PIRSF000337; NTA_MOA; 1.
DR SUPFAM; SSF51679; SSF51679; 1.
DR TIGRFAMs; TIGR03860; FMN_nitrolo; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Flavoprotein; FMN; Monooxygenase; Nucleotide-binding;
KW Oxidoreductase; Plasmid; Secreted.
FT CHAIN 1..440
FT /note="Long-chain alkane monooxygenase"
FT /id="PRO_0000454161"
FT BINDING 58
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:18164311,
FT ECO:0007744|PDB:3B9O"
FT BINDING 137..138
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:18164311,
FT ECO:0007744|PDB:3B9O"
FT BINDING 158
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:18164311,
FT ECO:0007744|PDB:3B9O"
FT BINDING 227..230
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:18164311,
FT ECO:0007744|PDB:3B9O"
FT MUTAGEN 14
FT /note="C->A: Loss of activity. Prevents dimerization."
FT /evidence="ECO:0000269|PubMed:18164311"
FT MUTAGEN 17
FT /note="H->F: Loss of activity. Can still form dimers."
FT /evidence="ECO:0000269|PubMed:18164311"
FT MUTAGEN 63
FT /note="Y->F: Loss of activity. Can still form dimers."
FT /evidence="ECO:0000269|PubMed:18164311"
FT MUTAGEN 79
FT /note="Q->L: Loss of activity. Can still form dimers."
FT /evidence="ECO:0000269|PubMed:18164311"
FT MUTAGEN 102
FT /note="A->D: 2.1-fold increase in activity toward
FT hexadecane. Uses a narrower spectrum of n-alkanes."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 102
FT /note="A->E: 2.2-fold increase in activity toward
FT hexadecane."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 146
FT /note="F->C: Loss of activity with hexadecane as substrate.
FT 2.9-fold increase in activity toward hexadecane; when
FT associated with I-376."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 146
FT /note="F->E: 2.0-fold increase in activity toward
FT hexadecane; when associated with I-376."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 146
FT /note="F->N: 3.4-fold increase in activity toward
FT hexadecane; when associated with I-376."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 146
FT /note="F->Q: 2.3-fold increase in activity toward
FT hexadecane; when associated with I-376."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 146
FT /note="F->R: 2.5-fold increase in activity toward
FT hexadecane; when associated with I-376."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 311
FT /note="H->F: Loss of activity. Can still form dimers."
FT /evidence="ECO:0000269|PubMed:18164311"
FT MUTAGEN 320
FT /note="L->A: 2.2-fold increase in activity toward
FT hexadecane. Uses a narrower spectrum of n-alkanes."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 320
FT /note="L->V: 2.5-fold increase in activity toward
FT hexadecane."
FT /evidence="ECO:0000269|PubMed:22526792"
FT MUTAGEN 376
FT /note="N->I: Loss of activity with hexadecane as substrate.
FT 2.9-fold increase in activity toward hexadecane; when
FT associated with C-146. 2.0-fold increase in activity toward
FT hexadecane; when associated with E-146. 3.4-fold increase
FT in activity toward hexadecane; when associated with N-146.
FT 2.3-fold increase in activity toward hexadecane; when
FT associated with Q-146. 2.5-fold increase in activity toward
FT hexadecane; when associated with R-146."
FT /evidence="ECO:0000269|PubMed:22526792"
FT STRAND 6..19
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 22..24
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 30..34
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 36..48
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 52..57
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 66..68
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 71..75
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 86..88
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 89..94
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 100..110
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 112..125
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 130..135
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 140..145
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 154..173
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 181..184
FT /evidence="ECO:0007829|PDB:3B9O"
FT TURN 185..188
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 189..191
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 193..195
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 218..221
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 223..227
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 231..240
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 242..246
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 248..250
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 251..265
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 266..268
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 272..274
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 275..285
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 289..302
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 305..316
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 320..322
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 328..333
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 334..341
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 347..354
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 356..370
FT /evidence="ECO:0007829|PDB:3B9O"
FT STRAND 374..378
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 384..399
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 412..417
FT /evidence="ECO:0007829|PDB:3B9O"
FT HELIX 429..433
FT /evidence="ECO:0007829|PDB:3B9O"
SQ SEQUENCE 440 AA; 50466 MW; DC04A40BDF8B06E4 CRC64;
MTKKIHINAF EMNCVGHIAH GLWRHPENQR HRYTDLNYWT ELAQLLEKGK FDALFLADVV
GIYDVYRQSR DTAVREAVQI PVNDPLMLIS AMAYVTKHLA FAVTFSTTYE HPYGHARRMS
TLDHLTKGRI AWNVVTSHLP SADKNFGIKK ILEHDERYDL ADEYLEVCYK LWEGSWEDNA
VIRDIENNIY TDPSKVHEIN HSGKYFEVPG PHLCEPSPQR TPVIYQAGMS ERGREFAAKH
AECVFLGGKD VETLKFFVDD IRKRAKKYGR NPDHIKMFAG ICVIVGKTHD EAMEKLNSFQ
KYWSLEGHLA HYGGGTGYDL SKYSSNDYIG SISVGEIINN MSKLDGKWFK LSVGTPKKVA
DEMQYLVEEA GIDGFNLVQY VSPGTFVDFI ELVVPELQKR GLYRVDYEEG TYREKLFGKG
NYRLPDDHIA ARYRNISSNV
