CNSA_STRCL
ID CNSA_STRCL Reviewed; 330 AA.
AC B5GMG2;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 14-OCT-2008, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=1,8-cineole synthase {ECO:0000303|PubMed:21726035};
DE Short=CnsA {ECO:0000303|PubMed:21726035};
DE Short=bCinS {ECO:0000303|PubMed:28966840};
DE EC=4.2.3.108 {ECO:0000269|PubMed:21726035, ECO:0000269|PubMed:28144299, ECO:0000269|PubMed:28966840};
DE AltName: Full=Monoterpene synthase {ECO:0000303|PubMed:28966840};
GN Name=cnsA {ECO:0000303|PubMed:21726035};
GN ORFNames=SCLAV_p0982 {ECO:0000312|EMBL:EFG04469.2},
GN SSCG_00536 {ECO:0000312|EMBL:EDY47508.1};
OS Streptomyces clavuligerus.
OG Plasmid pSCL4 {ECO:0000312|EMBL:EFG04469.2,
OG ECO:0000312|Proteomes:UP000002357}.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1901;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 27064 / DSM 738 / JCM 4710 / NBRC 13307 / NCIMB 12785 / NRRL
RC 3585 / VKM Ac-602 {ECO:0000312|Proteomes:UP000006569};
RA Fischbach M., Ward D., Young S., Jaffe D., Gnerre S., Berlin A., Heiman D.,
RA Hepburn T., Sykes S., Alvarado L., Kodira C.D., Straight P., Clardy J.,
RA Hung D., Kolter R., Mekalanos J., Walker S., Walsh C.T., Lander E.,
RA Galagan J., Nusbaum C., Birren B.;
RT "Annotation of Streptomyces clavuligerus ATCC 27064.";
RL Submitted (FEB-2008) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 27064 / DSM 738 / JCM 4710 / NBRC 13307 / NCIMB 12785 / NRRL
RC 3585 / VKM Ac-602 {ECO:0000312|Proteomes:UP000002357};
RC PLASMID=pSCL4 {ECO:0000312|EMBL:EFG04469.2};
RX PubMed=20624727; DOI=10.1093/gbe/evq013;
RA Medema M.H., Trefzer A., Kovalchuk A., van den Berg M., Mueller U.,
RA Heijne W., Wu L., Alam M.T., Ronning C.M., Nierman W.C., Bovenberg R.A.L.,
RA Breitling R., Takano E.;
RT "The sequence of a 1.8-mb bacterial linear plasmid reveals a rich
RT evolutionary reservoir of secondary metabolic pathways.";
RL Genome Biol. Evol. 2:212-224(2010).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=ATCC 27064 / DSM 738 / JCM 4710 / NBRC 13307 / NCIMB 12785 / NRRL
RC 3585 / VKM Ac-602;
RX PubMed=21726035; DOI=10.1002/cbic.201100330;
RA Nakano C., Kim H.K., Ohnishi Y.;
RT "Identification of the first bacterial monoterpene cyclase, a 1,8-cineole
RT synthase, that catalyzes the direct conversion of geranyl diphosphate.";
RL ChemBioChem 12:1988-1991(2011).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND REACTION MECHANISM.
RC STRAIN=ATCC 27064 / DSM 738 / JCM 4710 / NBRC 13307 / NCIMB 12785 / NRRL
RC 3585 / VKM Ac-602;
RX PubMed=28144299; DOI=10.3762/bjoc.12.225;
RA Rinkel J., Rabe P., Zur Horst L., Dickschat J.S.;
RT "A detailed view on 1,8-cineol biosynthesis by Streptomyces clavuligerus.";
RL Beilstein J. Org. Chem. 12:2317-2324(2016).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.51 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG AND
RP MAGMESIUM IONS, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE
RP SPECIFICITY, REACTION MECHANISM, DOMAIN, AND SUBUNIT.
RC STRAIN=ATCC 27064 / DSM 738 / JCM 4710 / NBRC 13307 / NCIMB 12785 / NRRL
RC 3585 / VKM Ac-602;
RX PubMed=28966840; DOI=10.1021/acscatal.7b01924;
RA Karuppiah V., Ranaghan K.E., Leferink N.G.H., Johannissen L.O.,
RA Shanmugam M., Cheallaigh A.N., Bennett N.J., Kearset L.J., Takano E.,
RA Gardiner J.M., van der Kamp M.W., Hay S., Mulholland A.J., Leys D.,
RA Scrutton N.S.;
RT "Structural basis of catalysis in the bacterial monoterpene synthases
RT linalool synthase and 1,8-cineole synthase.";
RL ACS Catal. 7:6268-6282(2017).
CC -!- FUNCTION: In vitro, catalyzes the formation of 1,8-cineole from geranyl
CC diphosphate (GPP) (PubMed:21726035, PubMed:28144299, PubMed:28966840).
CC Can also accept neryl diphosphate (NPP) as substrate to produce 1,8-
CC cineole (PubMed:28966840). {ECO:0000269|PubMed:21726035,
CC ECO:0000269|PubMed:28144299, ECO:0000269|PubMed:28966840}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E)-geranyl diphosphate + H2O = 1,8-cineole + diphosphate;
CC Xref=Rhea:RHEA:32543, ChEBI:CHEBI:15377, ChEBI:CHEBI:27961,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:58057; EC=4.2.3.108;
CC Evidence={ECO:0000269|PubMed:21726035, ECO:0000269|PubMed:28144299,
CC ECO:0000269|PubMed:28966840};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + neryl diphosphate = 1,8-cineole + diphosphate;
CC Xref=Rhea:RHEA:56632, ChEBI:CHEBI:15377, ChEBI:CHEBI:27961,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57665;
CC Evidence={ECO:0000269|PubMed:28966840};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:28966840};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000269|PubMed:28966840};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=170 nM for geranyl diphosphate (GPP)
CC {ECO:0000269|PubMed:21726035};
CC Note=kcat is 0.079 sec(-1) with geranyl diphosphate (GPP) as
CC substrate. {ECO:0000269|PubMed:21726035};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:28966840}.
CC -!- DOMAIN: The Asp-Asp-Xaa-Xaa-Asp (DDXXD) and Asn-Xaa-Xaa-Xaa-Ser-Xaa-
CC Xaa-Xaa-Glu (NSE) motifs are important for the catalytic activity,
CC presumably through binding to Mg(2+). {ECO:0000305|PubMed:28966840}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; DS570626; EDY47508.1; -; Genomic_DNA.
DR EMBL; CM000914; EFG04469.2; -; Genomic_DNA.
DR RefSeq; WP_003952918.1; NZ_CP032054.1.
DR PDB; 5NX6; X-ray; 1.63 A; A/B=1-330.
DR PDB; 5NX7; X-ray; 1.51 A; A/B=1-330.
DR PDBsum; 5NX6; -.
DR PDBsum; 5NX7; -.
DR AlphaFoldDB; B5GMG2; -.
DR SMR; B5GMG2; -.
DR STRING; 443255.SCLAV_p0982; -.
DR EnsemblBacteria; EDY47508; EDY47508; SSCG_00536.
DR GeneID; 61474073; -.
DR KEGG; sclf:BB341_29050; -.
DR eggNOG; COG0664; Bacteria.
DR OMA; PQLRVCC; -.
DR OrthoDB; 1869158at2; -.
DR BioCyc; MetaCyc:MON-17530; -.
DR BRENDA; 4.2.3.108; 5988.
DR Proteomes; UP000002357; Plasmid pSCL4.
DR Proteomes; UP000006569; Unassembled WGS sequence.
DR GO; GO:0102313; F:1,8-cineole synthase activity; IDA:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0033383; P:geranyl diphosphate metabolic process; IDA:UniProtKB.
DR Gene3D; 1.10.600.10; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR InterPro; IPR034686; Terpene_cyclase-like_2.
DR SFLD; SFLDG01020; Terpene_Cyclase_Like_2; 1.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Lyase; Magnesium; Metal-binding; Plasmid; Reference proteome.
FT CHAIN 1..330
FT /note="1,8-cineole synthase"
FT /id="PRO_0000445228"
FT MOTIF 81..85
FT /note="DDXXD motif"
FT /evidence="ECO:0000305|PubMed:28966840"
FT MOTIF 220..228
FT /note="NXXXSXXXE motif"
FT /evidence="ECO:0000305|PubMed:28966840"
FT BINDING 81
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT BINDING 81
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT BINDING 174
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX7"
FT BINDING 220
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT BINDING 224
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT BINDING 227
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT BINDING 228
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT BINDING 314..315
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:28966840,
FT ECO:0007744|PDB:5NX6, ECO:0007744|PDB:5NX7"
FT HELIX 20..33
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 40..48
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 53..58
FT /evidence="ECO:0007829|PDB:5NX7"
FT TURN 59..61
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 64..83
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 96..104
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 117..130
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 135..150
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 152..160
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 167..177
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 180..190
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 197..200
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 203..223
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 225..231
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 237..245
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 249..270
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 271..273
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 274..281
FT /evidence="ECO:0007829|PDB:5NX7"
FT HELIX 285..311
FT /evidence="ECO:0007829|PDB:5NX7"
SQ SEQUENCE 330 AA; 37725 MW; 05B46E3F7F34C372 CRC64;
MPAGHEEFDI PFPSRVNPFH ARAEDRHVAW MRAMGLITGD AAEATYRRWS PAKVGARWFY
LAQGEDLDLG CDIFGWFFAY DDHFDGPTGT DPRQTAAFVN RTVAMLDPRA DPTGEHPLNI
AFHDLWQRES APMSPLWQRR AVDHWTQYLT AHITEATNRT RHTSPTIADY LELRHRTGFM
PPLLDLIERV WRAEIPAPVY TTPEVQTLLH TTNQNINIVN DVLSLEKEEA HGDPHNLVLV
IQHERQSTRQ QALATARRMI DEWTDTFIRT EPRLPALCGR LGIPLADRTS LYTAVEGMRA
AIRGNYDWCA ETNRYAVHRP TGTGRATTPW
