Rapam_00130 : CDS information

Location

Organism	Streptomyces rapamycinicus
Strain	NRRL 5491
Entry name	Rapamycin
Contig	X86780
Start / Stop / Direction	67,319 / 71,944 / + [in whole cluster] 67,319 / 71,944 / + [in contig]
Location	67319..71944 [in whole cluster] 67319..71944 [in contig]
Type	CDS
Length	4,626 bp (1,541 aa)

Annotation

Category	1.2 NRPS
Product	non-ribosomal peptide synthetase
Product (GenBank)	pipecolate incorporating enzyme
Gene
Gene (GenBank)	rapP
EC number
Keyword
Note
Note (GenBank)
Reference	ACC Q54298 PmId [7644502] The biosynthetic gene cluster for the polyketide immunosuppressant rapamycin. (Proc Natl Acad Sci U S A. , 1995) [8635730] Organisation of the biosynthetic gene cluster for rapamycin in Streptomyces hygroscopicus: analysis of genes flanking the polyketide synthase. (Gene. , 1996) [9266694] The pipecolate-incorporating enzyme for the biosynthesis of the immunosuppressant rapamycin--nucleotide sequence analysis, disruption and heterologous expression of rapP from Streptomyces hygroscopicus. (Eur J Biochem. , 1997) comment [PMID: 7644502](1995) rapamycin生合成gene clusterの報告。配列解析。 RapP: pipecolate incorporating enzyme --- [PMID: 8635730](1996) PKS flanking genesの配列解析。 --- [PMID: 9266694](1997) putative pipecolate-incorporating enzymeをコードするrapPを同定、配列解析。 rapP破壊mutantでrapamycin非産生。 rapPをE.coli, S.coelicolorで発現、精製。 recombinant RapPの翻訳後修飾を確認。
Related Reference	ACC Q9KID8 NITE Asco_00160 PmId [15835888] Elucidating the substrate specificity and condensation domain activity of FkbP, the FK520 pipecolate-incorporating enzyme. (Biochemistry. , 2005) comment 2nd(Q9KID8)[Asco_00160] 59%, 0.0 Streptomyces hygroscopicus subsp. ascomyceticus_fkbP FkbP FkbPは4つのdomainのNRPS。 AA挿入の後、鎖を環化するのにもおそらく役立つ。 recombinant FkbPの発現と精製。FkbPはvirtoでphosphopantetheinylateされていたことを示し、そのPCP domainはL-pipecolic acidと共にうまくロードされた。 Site-directed mutagenesisで、N末adenylation domainがtransfer reactionを触媒することを確認。 FkbPのadenylation domainはpipecolate取り込みのための一次選択フィルターである。

PKS/NRPS Module

15		L-pipecolate
16

C	39..332

A	521..922

PCP	1007..1074

C	1092..1389

Sequence

>non-ribosomal peptide synthetase [pipecolate incorporating enzyme]
MASDSPRPRPLKPAFAVSRTQRQRRRLDERGISRADRNGPLPASFSQQRLWLMQQVAPDS
NSYNLPLVQRLRGALDTVILRRALGLVASRHEALRTVFDAEDGEPLQRIQPADVVRLREL
DARDEDHARALVRRETAEPFDLHAGPVMRALLVRLADDDHVLALTVHHIAGDGWSLAILR
MELSAQYAPLLRGAEADLPALPSQYADFAQWERKALSGTKLQKRLDYWQENLRDAPMVLG
LPADRPRPPVGSSDAGTVAWCPPTELVTAAKELSRSTGTTLFMSLLAAFQVVLSHYARTD
DVLVGTPIANRNHAEVEGLIGMFVNTVVLRGDLSGDPTFRELLLRTRTNAMGAFANAELP
FDLLVEKVAPQRDLSVNPVVQVLFQLMPLASNTFTLPGIAVEPFDMNQFFTRMDLEFHVY
EDPAGDRLTGEVWYSRALFDGPRIERMLDQFTLVLRSVLDNPDVPISQISLAPEIGTDLP
TVEANETDRDLPFDSLPELLADAAARNPDAIAVVDERVTLTYAELGGRANKMAHLLRHKG
VRPGELVGLCVDRGADMIRGHARILKAGAGYVPIDPEHPIERTRFGLDDSSISTGIAQET
YRTRFPDVRDIILPDDPGLENQPASPPDVTTDRNSLAYAIYTSGSTGRPKAVLMPGICVV
NLLLWQERTMGREPASRTAQFITATFDYSVQEIFSALLGGTLVIPPDDIRLDPARLAQWI
DDSRITRIYAPTTVLRALVEHVDPHGHGLVDIASPLPGGESLVLDGKLRECALHRPHLRV
HNHYGPAESQLVTGYTLPEDVSAWPSTTPIGKPIDNTRIHLLDDALRPYPTAYAAQVCIS
GIGLARGYLARPELTRQRFITEGTGSEPRMYLSGDLARRLPDGNLEFLGRIDDQVKIRGI
RIELGEIETALSEHAAITQAAVTVREDDRGDKRLVAYVVPHSDAGDLAVELRGHVEARLP
SYMVPSAFIVLDKLPLTTSGKTDRRALPAPESWSPGPTSPVSPRNATESTVCGIYADVLE
VQTVGVHDDFFALGGHSLLASRAVSRIRAELGCDLPLRTLFDVRTPALLAQAIGSMSSRD
MPVLAPSPRTTPAPISLAQQQLLGISDSLLDLGAFPVCPYGFRLRGEIDRTVLDTALTRI
VARHEPLRTGFRDDGDGFVQIVREPAPVRAEFIEVTGKDTTDREAAAAEIAKNELERPFD
LVDGILLRAVLLRLEQDDHILILMLHHIAGDGWSFDVLVRELSALYAECRVPVAGLTEVG
TTYSDFARWEHEALSGSIREEHDAYWREHLKGAVALELPTDRPRKLADPIGKSLEWTVSP
DATAAARRLAQAEGVTLYETMLGAFTLAAAGLSGLDDILVATPFANRGRPEIDHLIGFFA
KVAALRVDLSDDPTFREVLRRVQTAVLGAHAHQDLPYAAIQATATATATATATATASELP
PPLVQFQLISSLTSDLVLPGTTIERFGMVEADLGVGNGELAIWLFDDDSALHGTVVFDGS
LFDPARIQSLLSSVESTLHVVSATPSVRVSKAYAANGHARR

>non-ribosomal peptide synthetase [pipecolate incorporating enzyme]
ATGGCTTCCGACAGTCCCCGCCCAAGGCCGCTCAAACCGGCTTTCGCTGTTTCGCGGACG
CAGCGTCAGCGTCGCAGACTTGACGAGCGCGGCATCAGTCGCGCCGATCGGAACGGGCCA
CTTCCGGCGTCTTTCTCGCAGCAACGTCTATGGCTCATGCAGCAGGTCGCACCCGACAGC
AATTCCTATAATCTGCCGCTCGTGCAGCGGCTGCGGGGCGCCCTGGACACCGTGATCCTC
AGGCGCGCTCTCGGCCTCGTCGCAAGCCGCCACGAGGCGCTTCGGACCGTATTCGACGCG
GAGGACGGCGAGCCGCTCCAGCGGATCCAGCCCGCGGATGTCGTGCGCCTGCGCGAACTG
GACGCCCGGGACGAGGACCACGCCAGAGCGTTGGTCCGGCGGGAGACGGCCGAGCCGTTC
GACCTGCACGCCGGTCCTGTGATGAGAGCGTTGTTGGTACGACTCGCCGACGATGACCAC
GTGCTGGCGCTCACGGTGCATCACATCGCGGGCGACGGCTGGTCGCTGGCCATCCTGCGG
ATGGAGCTCTCGGCGCAGTACGCGCCGCTGCTTCGCGGTGCGGAGGCCGATCTGCCCGCA
CTCCCGTCGCAGTACGCGGACTTCGCCCAGTGGGAACGCAAGGCGTTGTCGGGCACCAAG
CTGCAAAAGCGGCTGGACTACTGGCAGGAGAACCTGCGGGACGCCCCGATGGTGCTGGGA
TTGCCGGCCGACCGGCCACGACCGCCGGTCGGCAGTTCGGACGCGGGCACGGTGGCCTGG
TGCCCGCCGACGGAATTGGTGACCGCGGCCAAAGAACTGAGCCGGTCGACGGGCACCACG
CTGTTCATGTCGCTGCTGGCCGCCTTCCAGGTGGTGCTGTCCCACTACGCGCGCACCGAT
GACGTCCTGGTCGGCACCCCGATCGCCAACCGCAATCACGCGGAGGTGGAGGGGCTGATC
GGCATGTTCGTGAACACGGTGGTGCTGCGCGGCGATCTGTCAGGTGATCCGACCTTCCGC
GAGCTCCTCCTGCGGACCCGTACGAACGCCATGGGCGCGTTCGCGAACGCCGAGCTGCCG
TTCGACCTCCTCGTCGAGAAGGTCGCACCGCAGCGCGACCTGTCGGTGAACCCGGTCGTG
CAGGTGCTCTTCCAGCTCATGCCGCTGGCGTCGAACACGTTTACGCTGCCGGGAATCGCG
GTCGAGCCGTTCGATATGAACCAGTTCTTCACCCGCATGGATCTGGAGTTCCATGTCTAC
GAGGATCCCGCCGGGGACAGGCTGACGGGCGAGGTCTGGTACAGCCGAGCGCTCTTCGAC
GGGCCTCGTATAGAACGAATGCTGGATCAGTTCACTCTCGTCCTGCGCAGCGTACTCGAC
AACCCGGACGTTCCGATCTCGCAGATTTCACTCGCGCCGGAGATCGGTACCGACCTGCCG
ACGGTCGAAGCGAACGAAACAGACCGAGACTTGCCGTTCGATTCGCTGCCGGAATTGCTG
GCGGACGCGGCAGCAAGAAACCCCGACGCGATCGCCGTCGTCGACGAACGGGTCACACTC
ACCTATGCCGAACTCGGTGGGCGGGCCAATAAGATGGCGCATCTGCTGCGACACAAAGGC
GTTCGCCCCGGCGAACTAGTCGGTCTTTGCGTCGACCGAGGCGCCGATATGATCCGTGGG
CATGCTCGGATTCTCAAAGCCGGCGCGGGCTATGTGCCCATCGACCCCGAGCATCCCATC
GAGCGCACCCGTTTCGGGTTGGACGATTCCAGCATATCCACGGGCATCGCTCAGGAGACG
TACCGGACTCGGTTCCCCGATGTCCGCGACATCATTCTCCCGGACGACCCCGGCCTCGAG
AATCAGCCCGCAAGCCCGCCCGATGTGACGACCGACCGCAACAGCCTGGCCTATGCGATT
TACACCTCCGGTTCCACGGGCCGTCCCAAGGCCGTGCTCATGCCCGGCATCTGCGTCGTC
AACCTGCTGCTGTGGCAGGAACGCACGATGGGTCGCGAACCGGCCAGTCGCACCGCCCAG
TTCATCACCGCGACCTTCGACTACTCGGTTCAGGAAATCTTCTCCGCGCTGCTCGGGGGC
ACGCTTGTCATTCCCCCCGACGACATACGGCTGGACCCCGCGCGACTCGCACAATGGATC
GACGACAGCCGTATCACGCGCATCTACGCACCCACCACGGTGCTGCGCGCGCTGGTCGAG
CACGTCGACCCGCACGGGCACGGGCTTGTCGACATTGCGTCACCTCTGCCAGGCGGCGAG
TCACTCGTGCTGGACGGCAAGTTACGAGAGTGTGCCCTGCACCGCCCGCATCTGCGGGTG
CACAACCACTATGGCCCGGCGGAGAGCCAACTCGTCACGGGCTACACACTGCCCGAGGAC
GTCTCCGCCTGGCCGTCCACGACCCCTATCGGGAAGCCGATCGACAACACGCGCATCCAC
CTTCTCGACGACGCGCTTCGTCCGTACCCGACGGCGTATGCGGCGCAGGTCTGCATATCG
GGGATCGGTCTGGCCCGCGGCTACCTGGCCCGGCCCGAACTGACCCGGCAGCGATTCATC
ACCGAAGGTACCGGCTCGGAGCCGAGGATGTACCTCTCCGGTGACCTGGCCCGGCGGCTG
CCGGACGGAAATCTCGAGTTTCTCGGGAGAATCGACGACCAGGTCAAGATCCGGGGCATC
CGGATCGAACTCGGCGAGATCGAGACGGCCCTGTCCGAACACGCGGCCATAACTCAGGCC
GCGGTGACCGTCCGAGAAGACGACCGCGGCGACAAACGGCTGGTCGCCTACGTGGTGCCC
CACTCGGATGCCGGGGACCTCGCCGTCGAATTGCGCGGTCATGTCGAGGCGCGGCTCCCC
TCTTACATGGTCCCGTCCGCGTTCATCGTCCTGGACAAGCTGCCGCTGACGACGAGCGGA
AAGACCGACCGGCGTGCGCTCCCCGCCCCGGAAAGCTGGTCACCGGGCCCCACCTCGCCA
GTCTCCCCGCGCAACGCGACCGAATCCACGGTTTGCGGGATCTACGCTGACGTGCTCGAG
GTGCAAACCGTCGGCGTGCACGACGATTTCTTCGCTCTCGGCGGCCACTCGCTGCTCGCC
TCCCGCGCCGTCTCGCGTATCCGTGCCGAGCTGGGCTGCGATCTACCGCTTCGTACGCTG
TTCGACGTCCGGACTCCCGCCTTGCTCGCCCAGGCCATCGGGTCGATGTCGTCGCGGGAC
ATGCCCGTGCTCGCGCCATCGCCGCGGACCACGCCCGCACCGATCTCGCTGGCCCAGCAG
CAGCTGCTCGGGATCAGTGATTCGCTGCTCGACCTGGGCGCATTTCCCGTCTGCCCGTAC
GGATTCCGGCTGCGCGGGGAGATCGACCGCACCGTACTCGACACCGCTCTGACCCGGATC
GTCGCGCGTCACGAGCCACTGCGAACAGGCTTCCGCGACGACGGCGACGGCTTCGTGCAG
ATCGTCCGCGAGCCCGCGCCGGTGAGGGCCGAGTTCATCGAGGTGACCGGGAAAGACACG
ACCGACCGTGAAGCGGCGGCGGCAGAGATCGCCAAGAACGAACTTGAACGTCCCTTCGAC
CTGGTCGACGGTATTCTGCTCCGCGCCGTTCTGCTGCGGCTGGAGCAAGACGACCACATC
CTCATCCTGATGTTGCACCACATCGCGGGCGACGGCTGGTCCTTCGACGTTCTGGTACGC
GAGCTGTCCGCGCTGTACGCCGAATGCCGCGTCCCCGTCGCGGGACTGACGGAAGTCGGC
ACGACATATTCGGACTTCGCCCGCTGGGAGCACGAGGCCCTGTCCGGGTCCATCCGCGAG
GAACACGACGCCTACTGGCGAGAGCACCTCAAGGGCGCTGTCGCACTGGAACTGCCCACC
GACAGGCCGCGAAAGCTTGCGGATCCGATCGGGAAATCCCTCGAGTGGACGGTTTCCCCG
GATGCGACAGCCGCAGCGCGGAGGCTCGCGCAGGCAGAGGGCGTGACCCTTTACGAAACC
ATGCTCGGAGCCTTCACGCTGGCCGCAGCCGGGTTGTCAGGCCTGGACGACATTCTGGTG
GCAACCCCTTTCGCCAATCGTGGCCGACCCGAGATCGACCACCTGATCGGCTTCTTCGCC
AAGGTTGCCGCCCTCCGTGTCGATCTCTCCGACGATCCGACGTTCCGCGAGGTCCTGCGA
CGCGTCCAGACCGCGGTCCTGGGTGCTCACGCGCACCAGGACCTCCCCTATGCGGCGATA
CAGGCCACGGCCACGGCCACGGCCACGGCCACGGCCACGGCCACGGCATCCGAACTCCCC
CCGCCGCTCGTGCAGTTCCAGCTGATCAGCTCGCTGACCTCGGATCTCGTCCTGCCTGGC
ACCACCATCGAGCGCTTCGGCATGGTCGAGGCCGATCTGGGGGTCGGGAACGGCGAACTC
GCCATCTGGCTCTTCGACGATGACAGCGCCCTGCACGGAACTGTCGTATTCGACGGATCC
CTATTCGACCCCGCGAGAATTCAGAGTCTGCTTTCCTCGGTCGAGTCGACGTTGCACGTT
GTCTCTGCCACCCCCTCCGTACGCGTATCCAAGGCATACGCAGCAAACGGCCACGCAAGG
AGATAG

[15] C	39..332

[15] A	521..922

[15] L-pipecolate	687..792

[15] PCP	1007..1074

[16] C	1092..1389

[15] C	115..996

[15] A	1561..2766

[15] L-pipecolate	2059..2376

[15] PCP	3019..3222

[16] C	3274..4167

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR000873 AMP-dependent synthetase/ligase (Domain) PF00501 AMP-binding IPR001242 Condensation domain (Domain) PF00668 Condensation IPR009081 Acyl carrier protein-like (Domain) G3DSA:1.10.1200.10 ACP_like SSF47336 ACP_like PF00550 PP-binding PS50075 ACP_DOMAIN IPR010071 Amino acid adenylation domain (Domain) TIGR01733 AA-adenyl-dom
SignalP	No significant hit
TMHMM	No significant hit

Rapam_00120 Rapam_00140