Chth_00050 Chth_00050   Chth_00070 Chth_00070

Chth_00060 : CDS information

close this sectionLocation

Organism
StrainDSM 40725
Entry nameChlorothricin
Contig
Start / Stop / Direction10,470 / 5,200 / - [in whole cluster]
10,470 / 5,200 / - [in contig]
Locationcomplement(5200..10470) [in whole cluster]
complement(5200..10470) [in contig]
TypeCDS
Length5,271 bp (1,756 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category1.1 PKS
Productpolyketide synthase
6-methylsalicylic acid synthase
Product (GenBank)ChlB1
Gene
Gene (GenBank)chlB1
EC number2.3.1.165
Keyword
  • iterative
  • 5-chloro-6-methyl-O-methylsalicylic acid
Note
Note (GenBank)
  • type I PKS
Reference
ACC
PmId
[16793515] Genetic characterization of the chlorothricin gene cluster as a model for spirotetronate antibiotic biosynthesis. (Chem Biol. , 2006)
[16677607] Cloning and characterization of a bacterial iterative type I polyketide synthase gene encoding the 6-methylsalicyclic acid synthase. (Biochem Biophys Res Commun. , 2006)
[20534347] Insights into bacterial 6-methylsalicylic acid synthase and its engineering to orsellinic acid synthase for spirotetronate generation. (Chem Biol. , 2010)
[26833898] Insights into 6-Methylsalicylic Acid Bio-assembly by Using Chemical Probes. (Angew Chem Int Ed Engl. , 2016)
comment
[PMID: 16793515](2006)
chlorothricin(CHL) gene clusterに関する文献。

chlB1(1,756 aa): Type I PKS: KS-AT-KR-DH-ACP

chlB1にはtype I PKS domainが存在することが示されており、chlB1の不活化変異株を作成し、その生産物をLC-MS, HR-MSを使って、CHLではなく、methylsalicylic acid moietyが欠損しているdesmethylsalicyloyl CHL(DM-CHL)であることを確認している。
また、このchlB1不活化変異株にchlB1遺伝子を相補すると、DM-CHLはCHLへと変換されることをHPLCで精製してLC-MSを使うことで確認している。

---
[PMID: 16677607](2006) abstract
6-methylsalicylic acid synthaseとして機能するiterative type I PKS gene chlB1のクローニング、配列解析、特徴づけ。

---
[PMID:20534347](2010)
ChlB1をS. albusで発現し、6-methylsalicylic acid(6-MSA)が発現されていることをHPLC-MSで確認。
site-specific mutagenesisによるChlB1のDH and KR domainsの機能的解析。

DH domain不活化株TL1072(H947A)は6-MSA非産生、(H947F)は少ないけど6-MSA産生あり。
ChlB1-DH domainのdehydration活性は重要だが、6-MSAを産生するのに不可欠でないかもしれない。

KR domain不活化株TL1076 (Y1540F) は、6-MSAのC-2が非還元のorsellinic acid蓄積。

---
[PMID: 26833898](2016)
6-methylsalicylic acid synthase による6-MSA生合成の中間体検出。
TH domainを介する経路の裏付けとなっている。
Related Reference
ACC
P87162
PmId
comment
BLAST id42%
Aspergillus terreus_atX
6-methylsalicylic acid synthase

6-methylsalicylic acid synthase_ATXのdehydratase(DH) domainの機能的検証。
DH domainはACPに繋がれたbeta-hydroxytriketide中間体のdehydrationに関連しないが、ACPから6-methylsalicylic acidを放出するためにthioester hydrolysisを触媒することが明らかになった。
よってDH domainからTH domain(Gly904 - Thr1048の145aa)へ改名。

構造解析報告のあるEryDH4とのalignmentから、H972XXXGXXXXP motif and Asp1129 が触媒に重要な残基であると提唱されている。著者らの既報で、ATX H972A mutantがbeta-hydroxy triketideのような産物をもたらさないことが確認されている。
ACC
P22367
PmId
[26833898] Insights into 6-Methylsalicylic Acid Bio-assembly by Using Chemical Probes. (Angew Chem Int Ed Engl. , 2016)
comment
BLAST id42%
Penicillium patulum (Penicillium griseofulvum)
6-methylsalicylic acid synthase (EC:2.3.1.165)

close this sectionPKS/NRPS Module

B1 acetyl-CoA
malonyl-CoA
KS17..391
AT556..871
TH879..1022
KR1380..1575
ACP1661..1734

close this sectionSequence

selected fasta
>polyketide synthase [ChlB1]
MQSHDVARAGGREVVEEPIAVLGMACRFAGGADTLEAFWELLLEGRDGIGEVPEKRWRAY
EEAGPDHAAAVRRATRWGGFLDDIEGFDAEFFGLSPREAELMDPQQRLLLEVAWEALEHA
GIAPRELAGTDAGVFVGIGSDDYGRRLLEDLPGIEAWTGIGSAMCAAANRISYALDLKGP
SLAVDTACSASLVAVHLACQSLRAGESEVSLAAGVNLMISPGLTLTLDAAGATAPDGRSK
SFDASADGYGRGEGCGLLVLKRLSDAVRDGDPVLAVIRGSSVNQDGKTNGIMAPSGSAQE
HVLDLACRRAGVDPASVDYVEAHGTGTRLGDPLEAGALSAVFGRGRPKDEPCLIGSVKSN
IGHLEAAAGIASLIKATLALSKGEIPPSLNFSQGNPAIDWAESGLRVVTERTAWPEREDR
PVRAGVSGFGYGGTIAHVVMEQAPEVSRPDDAAGDEGSAEVVTERLFPLSGGTQAGLRAY
AGRLADRLSDDDAEELPLESVGHTLALRRSALAHRAAVVASDRKDLVAKLRLITLGEQTR
EAVIGSVPSDAGAGPVWVFSGHGSQWSGMGRELLASEPAFAAVIDEIDPVFRAEIGFSAR
QALLDGDFDTVDRVQTMIFAVQVALAAVWHSYGAAPSAVIGHSVGEIAAAVAAGALSLTD
GARLICRRSRLLRRVAGQGAMAMASISFEEAAERLAGRTDVVPAIAASPLSAVVAGDPAA
INALIDEWQAQDIQMRRVASDVAFHSPHMDPLLTEIAAAAEDLTPRQPELPVYSTAMEDP
RSQATLDGSYWAANLRNPVRLQPAVTAAVEDGHRAFIEVSAHPVVTHSIGETLSELGQED
AFTGSSLRRNQPERATLLSAVGAAHCHGIAVDWARLHPTGDLVALPLVAWQRSPHWHERA
SAATGQGLQHDLDSHALLGPRVPVAGRPLELWRTLLDDETRPYPGSHTINGTEIVPAAVL
INTFLDAARAADGARPVLRDMALRLPLITTERRELQVVRDDNSLRLASRSLEDGAAWLTH
TTATAAPAGSGEALQDLAAGAVLRPADPGDVQRHLTSVGVPTMGFEWTIEELARSEGMLA
ARVSVERPQRAQETWAPLLDAALSIAPTAIPGPPALRMVASFEEIVTEGAPPAGPATIQV
AADPVHENTVDVRIADTDGQAVAWVRGLRYDGMDQGGMTAAHPRDLVFEMAWRPFEAPAP
QDVSARRIVLIAAHDVKPLRTALTRAGAHVDVGLDGTLDENTDVVVVPDLTADIPVPEAA
ARSAWLLLSTAQRIAALDTLRFPRLWCLTTAVRESQAETHLAQSTLWGLGRVIAGEHSEL
WGGVIDLAPGTPDATTLLSVLHTGGGEDVIALRDGTATTARLTTTQREPTGTPLECRADG
TYLITGGLGTLGLEVAGRLAERGARRLVLAGRTGLPPRSTWGETTDTHTRQRIEAVKALE
DQGVTVRVIPLDITDTAKAAEQLTPDALGLPPIRGIVHLAGVLDNRMVTAVDETSLRTVL
RPKADGAWTLHTLFPPGTIDFLILFSSCGQLLGLPGQAAYGSANAFLDALAVHRNTTTPT
AADTTSFGWTSWRGQGMAVNDVVDAELRARGVTDITTQEAFAAWDFAAQHGPGNYPVLRR
LPHEPDMDQLPLLSEIHHTQPTAPTSGAATDSYAGLAPDELRARLIDEVAAHISAEMKLA
ASQLDHRKSLVEQGLDSVMTIVIRRRLEKWFGHKLPATLLWHQPTVTAISEHLAELLAPT
TSQPDNTAPAEPAATA
selected fasta
>polyketide synthase [ChlB1]
GTGCAGAGTCACGACGTTGCCCGTGCGGGCGGCAGGGAAGTCGTCGAGGAGCCGATCGCC
GTGCTCGGGATGGCGTGCCGGTTCGCAGGTGGTGCCGACACCCTGGAGGCGTTCTGGGAG
TTGCTGCTGGAGGGCCGGGACGGCATCGGTGAGGTGCCTGAGAAGCGGTGGCGCGCCTAC
GAGGAGGCCGGCCCCGATCATGCGGCGGCGGTGCGGAGGGCGACGCGGTGGGGTGGGTTC
CTCGATGACATCGAGGGGTTCGACGCGGAGTTCTTCGGGTTGTCGCCGCGTGAGGCGGAG
TTGATGGATCCGCAGCAGCGGTTGCTGCTGGAGGTGGCGTGGGAGGCGTTGGAGCACGCG
GGTATTGCGCCGCGGGAGTTGGCGGGGACGGACGCGGGTGTGTTCGTGGGGATCGGTTCG
GATGATTACGGCCGGCGGTTGTTGGAGGATCTGCCGGGGATCGAGGCGTGGACGGGGATC
GGCAGTGCGATGTGTGCGGCGGCGAACCGGATCTCGTATGCGCTGGATCTGAAGGGGCCG
AGTCTGGCGGTGGACACGGCGTGTTCGGCGTCGTTGGTGGCGGTGCATCTGGCGTGTCAG
AGTCTGCGGGCGGGTGAGAGTGAGGTGTCGCTCGCGGCGGGTGTGAATCTGATGATCTCA
CCGGGGTTGACGCTGACGCTGGATGCGGCGGGTGCGACGGCGCCGGACGGGCGGTCGAAG
TCCTTCGATGCCTCCGCGGACGGTTATGGCCGGGGCGAGGGGTGTGGGCTGCTCGTGCTG
AAGCGGTTGTCGGACGCGGTGCGGGACGGGGATCCGGTGCTGGCGGTGATCCGGGGCAGT
TCGGTGAACCAGGACGGGAAGACGAACGGGATCATGGCGCCGAGTGGTTCGGCGCAGGAG
CATGTGCTGGATCTGGCGTGCCGGCGGGCGGGGGTGGATCCGGCGTCGGTGGATTACGTC
GAGGCGCATGGCACGGGGACGCGGCTTGGAGACCCGTTGGAAGCGGGTGCGCTGAGCGCG
GTGTTCGGGCGGGGGCGGCCCAAGGATGAGCCGTGTCTGATCGGTTCGGTGAAGTCGAAC
ATCGGGCATCTGGAGGCGGCGGCGGGGATTGCGAGCCTGATCAAGGCGACGCTGGCGTTG
AGCAAGGGAGAGATCCCGCCGAGTCTGAACTTCTCGCAGGGCAATCCGGCGATCGACTGG
GCGGAGTCCGGGCTGCGGGTGGTGACCGAGCGGACGGCCTGGCCCGAGCGGGAGGACCGA
CCGGTCCGTGCGGGCGTTTCCGGCTTCGGCTATGGCGGCACCATCGCGCATGTGGTCATG
GAGCAGGCGCCTGAGGTGAGTCGGCCCGATGACGCGGCGGGTGATGAGGGGTCTGCCGAG
GTCGTGACGGAGCGGCTGTTCCCGCTCTCGGGTGGAACGCAGGCCGGACTCCGGGCGTAT
GCGGGACGCCTCGCGGACCGGCTGTCGGACGACGACGCCGAGGAACTGCCCCTGGAGTCG
GTCGGGCACACCCTGGCCTTGCGCAGGTCGGCGCTGGCGCACCGGGCCGCCGTCGTGGCC
TCGGACCGCAAGGACCTGGTGGCCAAGCTGCGGTTGATCACGCTGGGGGAGCAGACCCGG
GAAGCCGTGATCGGGTCGGTACCCTCCGATGCCGGTGCGGGGCCGGTGTGGGTGTTCTCC
GGGCATGGTTCGCAGTGGTCGGGGATGGGGCGTGAACTGCTGGCGTCCGAGCCCGCGTTC
GCAGCGGTGATCGACGAGATCGATCCCGTTTTCCGTGCGGAGATCGGGTTCTCGGCCCGG
CAGGCTCTGCTCGACGGTGACTTCGACACCGTCGACCGTGTTCAGACGATGATTTTCGCG
GTGCAGGTCGCGCTGGCGGCGGTCTGGCACTCTTATGGTGCCGCCCCGTCGGCGGTGATC
GGGCACTCCGTGGGGGAGATCGCGGCGGCTGTGGCGGCGGGTGCGCTGTCGCTGACGGAC
GGAGCGCGGCTGATCTGCCGCCGCTCCCGACTCTTGCGGCGGGTGGCCGGCCAGGGAGCG
ATGGCTATGGCGAGCATCTCCTTCGAGGAGGCGGCCGAGCGGCTGGCGGGCCGTACGGAT
GTGGTGCCGGCGATTGCCGCGTCCCCGCTCTCCGCGGTCGTGGCAGGTGACCCTGCAGCG
ATCAACGCGCTGATCGACGAGTGGCAGGCACAGGACATCCAGATGCGCCGGGTCGCCTCG
GACGTGGCCTTCCACAGCCCGCACATGGACCCGCTGCTCACCGAAATCGCGGCCGCTGCC
GAGGACTTGACGCCGCGCCAGCCCGAACTCCCGGTGTACTCCACGGCCATGGAGGACCCC
CGCTCCCAGGCGACCCTCGACGGCTCCTACTGGGCCGCCAACCTGCGTAACCCGGTGCGG
TTGCAGCCGGCGGTGACGGCGGCGGTCGAGGACGGCCACCGCGCGTTCATCGAAGTGTCC
GCGCATCCCGTGGTCACGCACTCCATCGGCGAGACGCTCTCCGAGCTCGGCCAGGAGGAC
GCCTTCACCGGCTCCTCCCTGCGCCGCAACCAGCCCGAACGCGCCACCCTCCTGTCCGCC
GTCGGCGCGGCGCACTGCCATGGCATCGCGGTGGACTGGGCGCGTCTGCACCCGACCGGT
GACCTGGTCGCCCTGCCGCTGGTGGCCTGGCAGCGCAGCCCGCACTGGCACGAGCGGGCC
TCCGCCGCCACCGGCCAGGGCTTGCAGCACGACCTTGACTCCCACGCGCTGCTCGGGCCG
CGCGTCCCGGTCGCGGGACGGCCGCTGGAACTGTGGCGCACACTGCTCGACGACGAGACG
CGCCCCTACCCCGGCAGCCACACCATCAACGGCACGGAGATCGTGCCCGCCGCCGTCCTG
ATCAACACGTTCCTCGACGCGGCACGCGCCGCCGACGGGGCCCGCCCGGTCCTGCGGGAC
ATGGCGCTGCGGCTGCCGCTGATCACCACCGAGCGGCGCGAACTCCAGGTCGTCAGGGAC
GACAACTCCTTGCGTCTGGCCTCGCGTTCACTGGAGGACGGTGCCGCGTGGCTGACCCAC
ACCACCGCCACCGCCGCACCGGCGGGCAGCGGCGAAGCGCTCCAGGACCTGGCCGCCGGT
GCCGTGTTGCGCCCGGCGGACCCGGGTGATGTGCAGCGCCACCTGACCTCGGTGGGCGTG
CCGACCATGGGATTTGAGTGGACCATCGAGGAACTCGCCCGGAGCGAGGGCATGTTGGCC
GCACGTGTGAGTGTCGAGCGGCCGCAGCGGGCCCAGGAGACGTGGGCGCCCTTGCTGGAC
GCCGCGCTGTCCATCGCGCCGACGGCCATCCCCGGCCCGCCGGCCCTGCGCATGGTGGCC
TCCTTCGAGGAGATCGTCACCGAAGGCGCCCCGCCGGCCGGTCCGGCGACCATCCAGGTC
GCGGCCGACCCGGTCCACGAGAACACCGTCGACGTCCGGATCGCCGACACCGACGGGCAG
GCCGTGGCGTGGGTGCGCGGCCTGCGCTACGACGGCATGGACCAGGGCGGCATGACGGCG
GCGCACCCCCGCGACCTGGTCTTCGAGATGGCCTGGCGGCCCTTCGAGGCCCCCGCGCCG
CAGGACGTGTCCGCCCGCCGGATCGTCCTGATCGCCGCACACGACGTGAAGCCCCTGCGC
ACGGCCCTCACCCGTGCCGGCGCTCACGTCGACGTCGGGCTGGACGGCACGCTCGACGAG
AACACCGACGTCGTCGTGGTGCCCGACCTCACCGCGGACATCCCCGTCCCCGAGGCCGCA
GCCCGTTCCGCATGGCTGCTGCTGAGCACCGCGCAGCGCATCGCCGCCCTGGACACCCTG
CGCTTCCCCCGCCTGTGGTGCCTGACCACCGCAGTCCGTGAAAGCCAGGCCGAAACCCAC
CTCGCGCAGTCCACCCTGTGGGGCCTGGGCCGGGTGATCGCGGGCGAGCACAGCGAACTG
TGGGGCGGCGTCATCGACCTGGCCCCCGGCACCCCGGACGCCACCACCCTGCTCAGCGTC
CTGCACACCGGCGGCGGCGAGGACGTCATCGCCCTCCGCGACGGCACCGCCACCACGGCC
CGCCTCACCACGACGCAACGCGAGCCCACTGGCACCCCGCTGGAATGCCGGGCGGACGGA
ACGTACCTGATCACCGGCGGACTGGGCACCCTCGGCCTGGAAGTCGCCGGCCGGCTCGCC
GAACGCGGCGCCCGCCGTCTCGTCCTCGCCGGACGCACCGGACTGCCACCCCGCTCCACC
TGGGGCGAGACCACCGACACGCACACCAGGCAGCGCATCGAGGCCGTCAAGGCCCTCGAA
GACCAGGGCGTCACCGTCCGTGTCATCCCCCTCGACATCACCGACACGGCCAAGGCCGCC
GAACAGCTCACCCCCGACGCCCTGGGCCTGCCACCCATCCGCGGCATCGTCCACCTCGCC
GGCGTCCTCGACAACCGCATGGTGACCGCGGTCGACGAGACATCCCTGCGCACCGTGCTG
CGGCCCAAGGCCGACGGCGCCTGGACCCTGCACACCCTCTTCCCGCCCGGCACCATCGAC
TTCCTGATCCTGTTCTCCTCCTGCGGCCAGCTCCTCGGCCTGCCCGGCCAGGCCGCCTAC
GGCTCCGCCAACGCCTTCCTCGACGCCCTCGCCGTCCACCGCAACACCACCACCCCGACC
GCCGCCGACACCACCAGCTTCGGCTGGACCTCCTGGCGCGGCCAGGGCATGGCCGTCAAC
GACGTCGTCGACGCCGAACTGCGCGCCCGAGGCGTCACCGACATCACCACCCAGGAAGCC
TTCGCCGCCTGGGACTTCGCCGCACAACACGGCCCCGGAAACTACCCCGTCCTACGCCGG
CTGCCCCACGAGCCGGACATGGACCAGCTCCCCCTCCTCAGCGAGATCCACCACACCCAG
CCCACCGCCCCCACCTCCGGCGCCGCAACCGACTCCTACGCGGGCCTCGCCCCCGACGAA
CTGCGCGCCCGCCTCATCGACGAGGTCGCCGCACACATCTCGGCCGAGATGAAACTCGCC
GCCTCCCAGCTCGACCACCGCAAGTCCCTGGTCGAGCAGGGCCTGGACTCGGTGATGACG
ATCGTGATCCGGCGCCGCCTGGAGAAGTGGTTCGGTCACAAACTCCCCGCGACCCTGCTG
TGGCACCAGCCCACCGTCACCGCCATCAGCGAACACCTGGCCGAACTCCTGGCCCCCACC
ACGTCCCAGCCCGACAACACGGCACCCGCCGAACCGGCGGCAACGGCCTGA
[1] KS17..391
[1] AT556..871
[1] acetyl-CoA malonyl-CoA742..746
[1] TH879..1022
[1] KR1380..1575
[1] ACP1661..1734
[1] KS49..1173
[1] AT1666..2613
[1] acetyl-CoA malonyl-CoA2224..2238
[1] TH2635..3066
[1] KR4138..4725
[1] ACP4981..5202

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR001227 Acyl transferase domain (Domain)
[555-675] 2.59999999999995e-67 G3DSA:3.40.366.10 [742-860] 2.59999999999995e-67 G3DSA:3.40.366.10
G3DSA:3.40.366.10   Ac_transferase_reg
IPR009081 Acyl carrier protein-like (Domain)
[1668-1733] 1.2e-07 PF00550
PF00550   PP-binding
[1664-1738] 1.3e-13 G3DSA:1.10.1200.10
G3DSA:1.10.1200.10   ACP_like
[1658-1755] 7.30000590915208e-17 SSF47336
SSF47336   ACP_like
[1661-1734] PS50075
PS50075   ACP_DOMAIN
IPR013968 Polyketide synthase, KR (Domain)
[1380-1574] 5.69999999999998e-52 PF08659
PF08659   KR
IPR014030 Beta-ketoacyl synthase, N-terminal (Domain)
[17-266] 2.20000000000002e-93 PF00109
PF00109   ketoacyl-synt
IPR014031 Beta-ketoacyl synthase, C-terminal (Domain)
[274-391] 2.89999999999999e-45 PF02801
PF02801   Ketoacyl-synt_C
IPR014043 Acyl transferase (Domain)
[556-871] 4.69999999999992e-98 PF00698
PF00698   Acyl_transf_1
IPR016035 Acyl transferase/acyl hydrolase/lysophospholipase (Domain)
[554-837] 2.1000026783403e-61 SSF52151
SSF52151   Acyl_Trfase/lysoPlipase
IPR016036 Malonyl-CoA ACP transacylase, ACP-binding (Domain)
[677-741] 4.79999776836274e-10 SSF55048
SSF55048   Malonyl_transacylase_ACP-bd
IPR016038 Thiolase-like, subgroup (Domain)
[16-277] 8.80000000000003e-97 G3DSA:3.40.47.10 [278-445] 1.40000000000001e-63 G3DSA:3.40.47.10
G3DSA:3.40.47.10   Thiolase-like_subgr
IPR016039 Thiolase-like (Domain)
[16-445] 3.699978135861e-101 SSF53901
SSF53901   Thiolase-like
IPR016040 NAD(P)-binding domain (Domain)
[1379-1602] 2.4e-30 G3DSA:3.40.50.720
G3DSA:3.40.50.720   NAD(P)-bd
IPR018201 Beta-ketoacyl synthase, active site (Active_site)
[179-195] PS00606
PS00606   B_KETOACYL_SYNTHASE
IPR020801 Polyketide synthase, acyl transferase domain (Domain)
[558-851] 3.30001976117415e-95 SM00827
SM00827   PKS_AT
IPR020806 Polyketide synthase, phosphopantetheine-binding domain (Domain)
[1666-1737] 1.10000150671642e-19 SM00823
SM00823   PKS_PP
IPR020841 Polyketide synthase, beta-ketoacyl synthase domain (Domain)
[19-445] SM00825
SM00825   PKS_KS
IPR020842 Polyketide synthase/Fatty acid synthase, KR (Domain)
[1380-1575] 1.20000117458134e-48 SM00822
SM00822   PKS_KR
SignalP
[1-37] 0.149 Signal
Bacteria, Gram-positive   
TMHMM No significant hit
Chth_00050 Chth_00050   Chth_00070 Chth_00070
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