PCEST_PENRW
ID PCEST_PENRW Reviewed; 399 AA.
AC B6H6L7;
DT 17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT 16-DEC-2008, sequence version 1.
DT 03-AUG-2022, entry version 62.
DE RecName: Full=Lovastatin esterase {ECO:0000303|PubMed:28619444};
DE Short=PcEST {ECO:0000303|PubMed:28619444};
DE EC=3.1.1.- {ECO:0000269|PubMed:28619444};
GN Name=PcEST {ECO:0000303|PubMed:28619444};
GN ORFNames=Pc15g00720, PCH_Pc15g00720;
OS Penicillium rubens (strain ATCC 28089 / DSM 1075 / NRRL 1951 / Wisconsin
OS 54-1255) (Penicillium chrysogenum).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium;
OC Penicillium chrysogenum species complex.
OX NCBI_TaxID=500485;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 28089 / DSM 1075 / NRRL 1951 / Wisconsin 54-1255;
RX PubMed=18820685; DOI=10.1038/nbt.1498;
RA van den Berg M.A., Albang R., Albermann K., Badger J.H., Daran J.-M.,
RA Driessen A.J.M., Garcia-Estrada C., Fedorova N.D., Harris D.M.,
RA Heijne W.H.M., Joardar V.S., Kiel J.A.K.W., Kovalchuk A., Martin J.F.,
RA Nierman W.C., Nijland J.G., Pronk J.T., Roubos J.A., van der Klei I.J.,
RA van Peij N.N.M.E., Veenhuis M., von Doehren H., Wagner C., Wortman J.R.,
RA Bovenberg R.A.L.;
RT "Genome sequencing and analysis of the filamentous fungus Penicillium
RT chrysogenum.";
RL Nat. Biotechnol. 26:1161-1168(2008).
RN [2]
RP FUNCTION, SUBSTRATE SPECIFICITY, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND BIOTECHNOLOGY.
RX PubMed=28619444; DOI=10.1016/j.ymben.2017.06.005;
RA Huang X., Liang Y., Yang Y., Lu X.;
RT "Single-step production of the simvastatin precursor monacolin J by
RT engineering of an industrial strain of Aspergillus terreus.";
RL Metab. Eng. 42:109-114(2017).
RN [3] {ECO:0007744|PDB:6KJC, ECO:0007744|PDB:6KJD, ECO:0007744|PDB:6KJE, ECO:0007744|PDB:6KJF}
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF IN COMPLEX WITH SUBSTRATE,
RP FUNCTION, SUBSTRATE SPECIFICITY, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVE SITE, MUTAGENESIS OF SER-57; LYS-60; ASP-106; TYR-127;
RP TYR-170 AND TRP-344, AND BIOTECHNOLOGY.
RX PubMed=31839596; DOI=10.1074/jbc.ra119.011936;
RA Liang Y., Lu X.;
RT "Structural insights into the catalytic mechanism of lovastatin
RT hydrolase.";
RL J. Biol. Chem. 295:1047-1055(2020).
CC -!- FUNCTION: Esterase that can hydrolyze the side chain of lovastatin to
CC produce monacolin J (PubMed:28619444, PubMed:31839596). Is also able to
CC hydrolyze the side chains of mevastatin and pravastatin, but not
CC simvastatin (PubMed:28619444). {ECO:0000269|PubMed:28619444,
CC ECO:0000269|PubMed:31839596}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + lovastatin = (S)-2-methylbutanoate + H(+) + monacolin J;
CC Xref=Rhea:RHEA:62748, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:40303, ChEBI:CHEBI:79034, ChEBI:CHEBI:145932;
CC Evidence={ECO:0000269|PubMed:28619444};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62749;
CC Evidence={ECO:0000269|PubMed:28619444};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + pravastatin lactone = (S)-2-methylbutanoate + H(+) +
CC pravastatin diol lactone; Xref=Rhea:RHEA:62752, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:145931, ChEBI:CHEBI:145932,
CC ChEBI:CHEBI:145933; Evidence={ECO:0000269|PubMed:28619444};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62753;
CC Evidence={ECO:0000269|PubMed:28619444};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + mevastatin = (S)-2-methylbutanoate + compactin diol
CC lactone + H(+); Xref=Rhea:RHEA:62744, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:34652, ChEBI:CHEBI:34848,
CC ChEBI:CHEBI:145932; Evidence={ECO:0000269|PubMed:28619444};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62745;
CC Evidence={ECO:0000269|PubMed:28619444};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=39.4 uM for lovastatin (at 37 degrees Celsius)
CC {ECO:0000269|PubMed:28619444};
CC KM=5.46 uM for lovastatin (at 30 degrees Celsius)
CC {ECO:0000269|PubMed:31839596};
CC -!- BIOTECHNOLOGY: PcEST can be used to directly produce monacolin J
CC through single-step fermentation, which would be easily applied to
CC industrial production with the existing equipment and fermentation
CC process of lovastatin production and beneficial for the current
CC simvastatin industry and hypercholesterolemia market.
CC {ECO:0000269|PubMed:28619444, ECO:0000269|PubMed:31839596}.
CC -!- SIMILARITY: Belongs to the class-A beta-lactamase family.
CC {ECO:0000305}.
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DR EMBL; AM920430; CAP82958.1; -; Genomic_DNA.
DR RefSeq; XP_002560298.1; XM_002560252.1.
DR PDB; 6KJC; X-ray; 2.30 A; A/B=1-399.
DR PDB; 6KJD; X-ray; 2.30 A; A/B=1-399.
DR PDB; 6KJE; X-ray; 2.48 A; A/B=1-399.
DR PDB; 6KJF; X-ray; 2.40 A; A/B=1-399.
DR PDBsum; 6KJC; -.
DR PDBsum; 6KJD; -.
DR PDBsum; 6KJE; -.
DR PDBsum; 6KJF; -.
DR AlphaFoldDB; B6H6L7; -.
DR SMR; B6H6L7; -.
DR MEROPS; S12.950; -.
DR EnsemblFungi; CAP82958; CAP82958; PCH_Pc15g00720.
DR GeneID; 8309482; -.
DR KEGG; pcs:Pc15g00720; -.
DR VEuPathDB; FungiDB:PCH_Pc15g00720; -.
DR eggNOG; ENOG502S4UR; Eukaryota.
DR HOGENOM; CLU_020027_11_1_1; -.
DR OMA; GTFFWID; -.
DR OrthoDB; 1268012at2759; -.
DR BioCyc; PCHR:PC15G00720-MON; -.
DR Proteomes; UP000000724; Contig Pc00c15.
DR GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.40.710.10; -; 1.
DR InterPro; IPR001466; Beta-lactam-related.
DR InterPro; IPR012338; Beta-lactam/transpept-like.
DR Pfam; PF00144; Beta-lactamase; 1.
DR SUPFAM; SSF56601; SSF56601; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Reference proteome.
FT CHAIN 1..399
FT /note="Lovastatin esterase"
FT /id="PRO_0000449866"
FT ACT_SITE 57
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:31839596"
FT ACT_SITE 60
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:31839596"
FT ACT_SITE 170
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 57
FT /note="S->A: Abolishes the catalytic activity."
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 60
FT /note="K->A,H,Q,R,S: Abolishes the catalytic activity."
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 106
FT /note="D->A: Leads to improved solubility and
FT thermostability."
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 127
FT /note="Y->A: Abolishes the catalytic activity."
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 170
FT /note="Y->A,E,F,H: Abolishes the catalytic activity."
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 344
FT /note="W->A: Decreases the catalytic activity."
FT /evidence="ECO:0000269|PubMed:31839596"
FT MUTAGEN 344
FT /note="W->K: Abolishes the catalytic activity."
FT /evidence="ECO:0000269|PubMed:31839596"
FT HELIX 1..10
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 13..22
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 26..38
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 44..46
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 52..54
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 56..58
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 59..72
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 78..80
FT /evidence="ECO:0007829|PDB:6KJC"
FT TURN 83..85
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 87..90
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 94..98
FT /evidence="ECO:0007829|PDB:6KJD"
FT STRAND 104..107
FT /evidence="ECO:0007829|PDB:6KJD"
FT HELIX 115..119
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 128..130
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 132..140
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 152..155
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 160..162
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 173..185
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 189..196
FT /evidence="ECO:0007829|PDB:6KJC"
FT TURN 197..202
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 207..210
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 214..217
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 233..236
FT /evidence="ECO:0007829|PDB:6KJC"
FT TURN 242..245
FT /evidence="ECO:0007829|PDB:6KJC"
FT TURN 251..253
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 255..258
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 259..271
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 274..277
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 279..285
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 292..303
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 307..311
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 320..322
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 324..329
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 341..346
FT /evidence="ECO:0007829|PDB:6KJC"
FT TURN 347..349
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 350..355
FT /evidence="ECO:0007829|PDB:6KJC"
FT TURN 356..359
FT /evidence="ECO:0007829|PDB:6KJC"
FT STRAND 360..368
FT /evidence="ECO:0007829|PDB:6KJC"
FT HELIX 374..396
FT /evidence="ECO:0007829|PDB:6KJC"
SQ SEQUENCE 399 AA; 43292 MW; DBA944027F39416F CRC64;
MDTTFQAAID TGKINGAVVC ATDAQGHFVY NKATGERTLL SGEKQPQQLD DVLYLASATK
LITTIAALQC VEDGLLSLDG DLSSIAPELA AKYVLTGFTD DESPLDDPPA RPITLKMLLT
HSSGTSYHFL DPSIAKWRAQ YANPENEKPR LVEEMFTYPL SFQPGTGWMY GPGLDWAGRV
VERVTGGTLM EFMQKRIFDP LGITDSQFYP VTREDLRARL VDLNPSDPGA LGSAVIGGGG
EMNLRGRGAF GGHGLFLTGL DFVKILRSLL ANDGMLLKPA AVDNMFQQHL GPEAAASHRA
ALASPLGPFF RVGTDPETKV GYGLGGLLTL EDVDGWYGER TLTWGGGLTL TWFIDRKNNL
CGVGAIQAVL PVDGDLMADL KQTFRHDIYR KYSAWKGQQ
