PENA_STREX
ID PENA_STREX Reviewed; 337 AA.
AC Q55012; E3VWK6;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 4.
DT 03-AUG-2022, entry version 101.
DE RecName: Full=Pentalenene synthase;
DE Short=PS;
DE EC=4.2.3.7;
DE AltName: Full=Pentalenolactone biosynthesis protein A;
DE AltName: Full=Sesquiterpene cyclase;
DE AltName: Full=Sesquiterpene synthase;
GN Name=penA;
OS Streptomyces exfoliatus (Streptomyces hydrogenans).
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1905;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-28; 57-73; 96-120
RP AND 145-157, FUNCTION, SUBUNIT, COFACTOR, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RC STRAIN=UC5319;
RX PubMed=8180213; DOI=10.1021/bi00185a024;
RA Cane D.E., Sohng J.-K., Lamberson C.R., Rudnicki S.M., Wu Z., Lloyd M.D.,
RA Oliver J.S., Hubbard B.R.;
RT "Pentalenene synthase. Purification, molecular cloning, sequencing, and
RT high-level expression in Escherichia coli of a terpenoid cyclase from
RT Streptomyces UC5319.";
RL Biochemistry 33:5846-5857(1994).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=UC5319;
RX PubMed=21284395; DOI=10.1021/ja111279h;
RA Zhu D., Seo M.J., Ikeda H., Cane D.E.;
RT "Genome mining in streptomyces. Discovery of an unprecedented P450-
RT catalyzed oxidative rearrangement that is the final step in the
RT biosynthesis of pentalenolactone.";
RL J. Am. Chem. Soc. 133:2128-2131(2011).
RN [3]
RP MUTAGENESIS OF HIS-309.
RX DOI=10.1021/ja983657h;
RA Seemann M., Zhai G., Umezawa K., Cane D.E.;
RT "Pentalenene synthase. Histidine-309 is not required for catalytic
RT activity.";
RL J. Am. Chem. Soc. 121:591-592(1999).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS).
RX PubMed=9295272; DOI=10.1126/science.277.5333.1820;
RA Lesburg C.A., Zhai G., Cane D.E., Christianson D.W.;
RT "Crystal structure of pentalenene synthase: mechanistic insights on
RT terpenoid cyclization reactions in biology.";
RL Science 277:1820-1824(1997).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.9 ANGSTROMS) OF MUTANT LEU-219, CATALYTIC
RP ACTIVITY, AND MUTAGENESIS OF PHE-76; PHE-77; ASP-80; ASP-81; ASP-84;
RP ASN-219; 308-TRP-HIS-309; TRP-308 AND HIS-309.
RX PubMed=12083921; DOI=10.1021/ja026058q;
RA Seemann M., Zhai G., de Kraker J.-W., Paschall C.M., Christianson D.W.,
RA Cane D.E.;
RT "Pentalenene synthase. Analysis of active site residues by site-directed
RT mutagenesis.";
RL J. Am. Chem. Soc. 124:7681-7689(2002).
CC -!- FUNCTION: Catalyzes the cyclization of farnesyl diphosphate (FPP) to
CC the tricyclic sesquiterpene pentalenene, which is the hydrocarbon
CC precursor of the pentalenolactone family of antibiotics produced by a
CC variety of Streptomyces species. {ECO:0000269|PubMed:8180213}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E)-farnesyl diphosphate = diphosphate + pentalenene;
CC Xref=Rhea:RHEA:18081, ChEBI:CHEBI:17251, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:175763; EC=4.2.3.7;
CC Evidence={ECO:0000269|PubMed:12083921};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000250};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=310 nM for FPP {ECO:0000269|PubMed:8180213};
CC pH dependence:
CC Optimum pH is 8.2-8.4. {ECO:0000269|PubMed:8180213};
CC -!- PATHWAY: Sesquiterpene biosynthesis; pentalenene biosynthesis;
CC pentalenene from farnesyl diphosphate: step 1/1.
CC -!- PATHWAY: Antibiotic biosynthesis; pentalenolactone biosynthesis.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:8180213}.
CC -!- DOMAIN: The Asp-Asp-Xaa-Xaa-Asp/Glu (DDXXD/E) motif is important for
CC the catalytic activity, presumably through binding to Mg(2+).
CC -!- MISCELLANEOUS: All the mutants (Ref.3 and PubMed:12083921) retained
CC substantial pentalenene synthase activity accompanied by production of
CC varying proportions of abortive cyclization products such as germacrene
CC A, protoilludene and beta-caryophyllene. This is a true derailment or
CC diversion of the normal cyclization reaction, and not simply the
CC consequence of trapping of a normally cryptic, cationic intermediate.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; U05213; AAA19131.1; -; Unassigned_DNA.
DR EMBL; HQ292066; ADO85594.1; -; Genomic_DNA.
DR PDB; 1HM4; X-ray; 3.47 A; A/B=2-337.
DR PDB; 1HM7; X-ray; 2.90 A; A/B=2-337.
DR PDB; 1PS1; X-ray; 2.60 A; A/B=1-337.
DR PDB; 6WKC; X-ray; 1.65 A; A/B=1-337.
DR PDB; 6WKD; X-ray; 2.20 A; A/B=1-337.
DR PDB; 6WKE; X-ray; 2.40 A; A/B=1-337.
DR PDB; 6WKF; X-ray; 2.50 A; A/B=1-337.
DR PDB; 6WKG; X-ray; 2.30 A; A/B=1-337.
DR PDB; 6WKH; X-ray; 2.55 A; A/B=1-337.
DR PDB; 6WKI; X-ray; 2.35 A; A/B=1-337.
DR PDB; 6WKJ; X-ray; 2.30 A; A/B=1-337.
DR PDBsum; 1HM4; -.
DR PDBsum; 1HM7; -.
DR PDBsum; 1PS1; -.
DR PDBsum; 6WKC; -.
DR PDBsum; 6WKD; -.
DR PDBsum; 6WKE; -.
DR PDBsum; 6WKF; -.
DR PDBsum; 6WKG; -.
DR PDBsum; 6WKH; -.
DR PDBsum; 6WKI; -.
DR PDBsum; 6WKJ; -.
DR AlphaFoldDB; Q55012; -.
DR SMR; Q55012; -.
DR BioCyc; MetaCyc:MON-16834; -.
DR BRENDA; 4.2.3.7; 6010.
DR UniPathway; UPA00171; UER00581.
DR UniPathway; UPA00974; -.
DR EvolutionaryTrace; Q55012; -.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0050467; F:pentalenene synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
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; Antibiotic biosynthesis; Direct protein sequencing;
KW Disulfide bond; Lyase; Magnesium; Metal-binding.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:8180213"
FT CHAIN 2..337
FT /note="Pentalenene synthase"
FT /id="PRO_0000097092"
FT MOTIF 80..84
FT /note="DDXXD motif"
FT BINDING 80
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT BINDING 80
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT BINDING 84
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT BINDING 84
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT BINDING 219
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT BINDING 223
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250"
FT BINDING 227
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250"
FT DISULFID 128..136
FT MUTAGEN 76
FT /note="F->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 77
FT /note="F->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 77
FT /note="F->Y: 20-fold decrease in activity and catalytic
FT efficiency."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 80
FT /note="D->E: 150-fold decrease in activity. 20-fold
FT increase in Km for FPP."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 81
FT /note="D->E: 50-fold decrease in activity. 9-fold increase
FT in Km for FPP."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 84
FT /note="D->E: 2.5-fold increase in activity. 7-fold increase
FT in Km for FPP."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 219
FT /note="N->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 219
FT /note="N->D: 60-fold decrease in activity. 55-fold increase
FT in Km for FPP."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 219
FT /note="N->L: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 308..309
FT /note="WH->FF: 12-fold decrease in activity. 25-fold
FT decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 308
FT /note="W->F: 4-fold decrease in activity. 2-fold decrease
FT in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12083921"
FT MUTAGEN 309
FT /note="H->A: 3-fold decrease in activity."
FT /evidence="ECO:0000269|PubMed:12083921, ECO:0000269|Ref.3"
FT MUTAGEN 309
FT /note="H->C: 4-fold decrease in activity."
FT /evidence="ECO:0000269|PubMed:12083921, ECO:0000269|Ref.3"
FT MUTAGEN 309
FT /note="H->F: 17-fold decrease in activity. 5-fold increase
FT in Km for FPP."
FT /evidence="ECO:0000269|PubMed:12083921, ECO:0000269|Ref.3"
FT MUTAGEN 309
FT /note="H->S: 3-fold decrease in activity."
FT /evidence="ECO:0000269|PubMed:12083921, ECO:0000269|Ref.3"
FT CONFLICT 118
FT /note="H -> W (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT HELIX 19..24
FT /evidence="ECO:0007829|PDB:6WKC"
FT TURN 25..27
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 28..32
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 39..47
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 50..57
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 63..81
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 86..89
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 91..102
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 103..105
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 115..128
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 133..158
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 166..176
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 179..190
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 196..199
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 202..222
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 224..229
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 236..244
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 248..271
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 273..279
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 284..296
FT /evidence="ECO:0007829|PDB:6WKC"
FT HELIX 298..308
FT /evidence="ECO:0007829|PDB:6WKC"
FT TURN 309..311
FT /evidence="ECO:0007829|PDB:1PS1"
SQ SEQUENCE 337 AA; 38002 MW; 417FCE6D8C4BB18C CRC64;
MPQDVDFHIP LPGRQSPDHA RAEAEQLAWP RSLGLIRSDA AAERHLRGGY ADLASRFYPH
ATGADLDLGV DLMSWFFLFD DLFDGPRGEN PEDTKQLTDQ VAAALDGPLP DTAPPIAHGF
ADIWRRTCEG MTPAWCARSA RHWRNYFDGY VDEAESRFWN APCDSAAQYL AMRRHTIGVQ
PTVDLAERAG RFEVPHRVFD SAVMSAMLQI AVDVNLLLND IASLEKEEAR GEQNNMVMIL
RREHGWSKSR SVSHMQNEVR ARLEQYLLLE SCLPKVGEIY QLDTAEREAL ERYRTDAVRT
VIRGSYDWHR SSGRYDAEFA LAAGAQGYLE ELGSSAH
