LSD19_STRLS
ID LSD19_STRLS Reviewed; 282 AA.
AC B6ZK72; B5M9L7;
DT 16-MAY-2012, integrated into UniProtKB/Swiss-Prot.
DT 10-FEB-2009, sequence version 1.
DT 03-AUG-2022, entry version 27.
DE RecName: Full=Epoxide hydrolase LasB;
DE EC=5.5.1.-;
DE AltName: Full=Lasalocid biosynthesis protein Lsd19;
GN Name=lsd19; Synonyms=lasB;
OS Streptomyces lasalocidi (Streptomyces lasaliensis).
OG Plasmid pKSL.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=324833;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION IN LASALOCID BIOSYNTHESIS, AND
RP DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 35851 / NRRL 3382R;
RX PubMed=19025863; DOI=10.1002/cbic.200800585;
RA Smith L., Hong H., Spencer J.B., Leadlay P.F.;
RT "Analysis of specific mutants in the lasalocid gene cluster: evidence for
RT enzymatic catalysis of a disfavoured polyether ring closure.";
RL ChemBioChem 9:2967-2975(2008).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 35851 / NRRL 3382R; PLASMID=pKSL;
RX PubMed=19129623; DOI=10.1271/bbb.80631;
RA Migita A., Watanabe M., Hirose Y., Watanabe K., Tokiwano T., Kinashi H.,
RA Oikawa H.;
RT "Identification of a gene cluster of polyether antibiotic lasalocid from
RT Streptomyces lasaliensis.";
RL Biosci. Biotechnol. Biochem. 73:169-176(2009).
RN [3]
RP FUNCTION AS AN EPOXIDE HYDROLASE, AND CATALYTIC ACTIVITY.
RC STRAIN=ATCC 35851 / NRRL 3382R; PLASMID=pKSL;
RX PubMed=18710235; DOI=10.1021/ja8040543;
RA Shichijo Y., Migita A., Oguri H., Watanabe M., Tokiwano T., Watanabe K.,
RA Oikawa H.;
RT "Epoxide hydrolase Lsd19 for polyether formation in the biosynthesis of
RT lasalocid A: direct experimental evidence on polyene-polyepoxide hypothesis
RT in polyether biosynthesis.";
RL J. Am. Chem. Soc. 130:12230-12231(2008).
RN [4]
RP FUNCTION, AND SUBSTRATE SPECIFICITY.
RX PubMed=20394359; DOI=10.1021/ol100541e;
RA Matsuura Y., Shichijo Y., Minami A., Migita A., Oguri H., Watanabe M.,
RA Tokiwano T., Watanabe K., Oikawa H.;
RT "Intriguing substrate tolerance of epoxide hydrolase Lsd19 involved in
RT biosynthesis of the ionophore antibiotic lasalocid A.";
RL Org. Lett. 12:2226-2229(2010).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM, DOMAIN, AND MUTAGENESIS
RP OF ASP-38; GLU-65; ASP-170 AND GLU-197.
RX PubMed=21375229; DOI=10.1021/ol200100e;
RA Minami A., Migita A., Inada D., Hotta K., Watanabe K., Oguri H., Oikawa H.;
RT "Enzymatic epoxide-opening cascades catalyzed by a pair of epoxide
RT hydrolases in the ionophore polyether biosynthesis.";
RL Org. Lett. 13:1638-1641(2011).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.59 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND
RP PRODUCT ANALOG, DOMAIN, REACTION MECHANISM, AND ACTIVE SITES.
RX PubMed=22388816; DOI=10.1038/nature10865;
RA Hotta K., Chen X., Paton R.S., Minami A., Li H., Swaminathan K.,
RA Mathews I.I., Watanabe K., Oikawa H., Houk K.N., Kim C.Y.;
RT "Enzymatic catalysis of anti-Baldwin ring closure in polyether
RT biosynthesis.";
RL Nature 483:355-358(2012).
CC -!- FUNCTION: Epoxide hydrolase responsible for the double epoxide-opening
CC cyclization of bisepoxyprelasalocid A to form lasalocid A, a polyether
CC antibiotic. In vitro, accepts various substrate analogs differing in
CC the left segment of lasalocid and epoxide stereochemistry to afford
CC products with excellent regioselectivity. {ECO:0000269|PubMed:18710235,
CC ECO:0000269|PubMed:19025863, ECO:0000269|PubMed:20394359,
CC ECO:0000269|PubMed:21375229}.
CC -!- DOMAIN: Consists of two highly homologous domains, oriented in a head-
CC to-tail fashion, that catalyze epoxide-opening cascades independently
CC in lasalocid biosynthesis. The N-terminal domain (Lsd19A) recognizes
CC the internal C18-C19 epoxide of the substrate and catalyzes
CC energetically favored 5-exo cyclization, while the C-terminal domain
CC (Lsd19B) recognizes the internal C22-C23 epoxide of the substrate and
CC predominantly catalyzes energetically disfavored 6-endo cyclization.
CC {ECO:0000269|PubMed:21375229, ECO:0000269|PubMed:22388816}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene do not produce lasalocid,
CC they only produce iso-lasalocid. {ECO:0000269|PubMed:19025863}.
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DR EMBL; FM173265; CAQ64695.1; -; Genomic_DNA.
DR EMBL; AB449340; BAG85034.1; -; Genomic_DNA.
DR PDB; 3RGA; X-ray; 1.59 A; A=1-282.
DR PDB; 4RZM; X-ray; 2.33 A; A/B=1-280.
DR PDBsum; 3RGA; -.
DR PDBsum; 4RZM; -.
DR AlphaFoldDB; B6ZK72; -.
DR SMR; B6ZK72; -.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR InterPro; IPR032710; NTF2-like_dom_sf.
DR InterPro; IPR037401; SnoaL-like.
DR Pfam; PF12680; SnoaL_2; 2.
DR SUPFAM; SSF54427; SSF54427; 2.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Isomerase; Plasmid.
FT CHAIN 1..282
FT /note="Epoxide hydrolase LasB"
FT /id="PRO_0000417070"
FT REGION 1..133
FT /note="Lsd19A"
FT REGION 134..282
FT /note="Lsd19B"
FT ACT_SITE 38
FT /note="Proton acceptor; for 5-exo epoxide-opening
FT cyclization activity"
FT /evidence="ECO:0000269|PubMed:22388816"
FT ACT_SITE 170
FT /note="Proton acceptor; for 6-endo epoxide-opening
FT cyclization activity"
FT /evidence="ECO:0000269|PubMed:22388816"
FT BINDING 14
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22388816"
FT BINDING 65
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22388816"
FT BINDING 146
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22388816"
FT BINDING 177
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22388816"
FT BINDING 197
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22388816"
FT BINDING 251
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22388816"
FT SITE 54
FT /note="Raises pKa of active site Asp-38"
FT SITE 186
FT /note="Raises pKa of active site Asp-170"
FT MUTAGEN 38
FT /note="D->A: Loss of both epoxide-opening activities; when
FT associated with A-197."
FT /evidence="ECO:0000269|PubMed:21375229"
FT MUTAGEN 65
FT /note="E->A: Loss of both epoxide-opening activities; when
FT associated with A-197."
FT /evidence="ECO:0000269|PubMed:21375229"
FT MUTAGEN 170
FT /note="D->A: Retains the 5-exo epoxide-opening activity but
FT loses the 6-endo epoxide-opening activity."
FT /evidence="ECO:0000269|PubMed:21375229"
FT MUTAGEN 197
FT /note="E->A: Retains the 5-exo epoxide-opening activity but
FT loses the 6-endo epoxide-opening activity. Loss of both
FT epoxide-opening activities; when associated with A-38 or
FT with A-65."
FT /evidence="ECO:0000269|PubMed:21375229"
FT CONFLICT 35
FT /note="L -> R (in Ref. 1; CAQ64695)"
FT /evidence="ECO:0000305"
FT HELIX 4..20
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 23..28
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 30..37
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 39..42
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 45..47
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 48..60
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 64..67
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 74..87
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 94..96
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 98..108
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 114..120
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 123..125
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 134..151
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 155..159
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 162..171
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 176..179
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 180..192
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 196..204
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 208..221
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 224..230
FT /evidence="ECO:0007829|PDB:3RGA"
FT TURN 242..244
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 246..257
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 263..269
FT /evidence="ECO:0007829|PDB:3RGA"
FT HELIX 272..274
FT /evidence="ECO:0007829|PDB:3RGA"
FT STRAND 275..278
FT /evidence="ECO:0007829|PDB:3RGA"
SQ SEQUENCE 282 AA; 30249 MW; F5F87F19197D15B7 CRC64;
MPAETVRKEV ALEYCRRVNA GELEGVLQLF APDALLVDPL GTEPVVGRAA LAARLAPALR
GAVHEEPGRP YAAHDGTSVV LPATVTVGAP GAPPQRRGRT RVMGVIEVGE DGLIREMRVM
WGVTDSSWTA RPAPDEERRK ELAREHCLRI NDGDVDGLLK LYSPRIRFED PVGSWTRTGL
EALRAHATMA VGSNVRETAG LTVAGQDGRH AAVTVSATMD YLPSGPLLAR HHLMTLPAPA
DPHRALIGIE YVMVIGVDAD GLIDEMRAYW GATDVSLLDP AA
