Balhi_00110 : CDS information

Location

Organism	Amycolatopsis balhimycina
Strain	DSM 5908
Entry name	Balhimycin
Contig	Y16952
Start / Stop / Direction	34,782 / 35,957 / + [in whole cluster] 34,782 / 35,957 / + [in contig]
Location	34782..35957 [in whole cluster] 34782..35957 [in contig]
Type	CDS
Length	1,176 bp (391 aa)

Annotation

Category	3.3 modification reduction
Product	cytochrome P450
Product (GenBank)	P450 monooxygenase
Gene
Gene (GenBank)	oxyA
EC number
Keyword	2nd cross-linking D-O-E ring
Note
Note (GenBank)	already deposited under Y16952
Reference	ACC O87673 PmId [10390204] Identification and analysis of the balhimycin biosynthetic gene cluster and its use for manipulating glycopeptide biosynthesis in Amycolatopsis mediterranei DSM5908. (Antimicrob Agents Chemother. , 1999) [11353482] The Biosynthesis of Vancomycin-Type Glycopeptide Antibiotics-New Insights into the Cyclization Steps This work was supported by the Deutsche Forschungsgemeinschaft (SFB 323). We thank M. Schierle, Dr. S. Stevanovic and Prof. H.-G. Rammensee for help with Edman degradation and J. Turner, Prof. B. List and Prof. D. Boger (La Jolla, USA) for discussions on the work. (Angew Chem Int Ed Engl. , 2001) [12404385] The Biosynthesis of Vancomycin-Type Glycopeptide Antibiotics-The Order of the Cyclization Steps This work was supported by the Deutsche Forschungsgemeinschaft (SFB 323) and by a grant of the EU (MEGATOP, QLK3-1999-00650). R. D. S. gratefully acknowledges the support of a Feodor-Lynen Fellowship granted by the Alexander-von-Humboldt Stiftung. We thank Corina Bihlmaier and Volker Pfeifer for help with transformation and Southern hybridization, J. A. Moss (La Jolla (USA)) for critical comments on the manuscript and Prof. Dr. M. E. Maier and Prof. Dr. H.-P. Fiedler (Tubingen) for generous support. (Angew Chem Int Ed Engl. , 2001) [15651041] The biosynthesis of vancomycin-type glycopeptide antibiotics--a model for oxidative side-chain cross-linking by oxygenases coupled to the action of peptide synthetases. (Chembiochem. , 2005) [16730832] Genetic analysis of the balhimycin (vancomycin-type) oxygenase genes. (J Biotechnol. , 2006) comment [PMID: 10390204](1999) Amycolatopsis mediterranei DSM5908からグリコペプチド系抗生物質balhimycinの生合成に関する9,882bpのDNA断片を同定。そこにoxyA-bgtfCまで7つのcomplete genesを同定した。 oxyA-Cの推定産物はcytochrome P450 monooxygenasesへの有意な配列類似を示す。 putative oxygenase gene(oxyA_C末-oxyB_N末)の不活化株SP1-1は抗菌的に活性のある化合物を合成できなかった。 --- [PMID: 11353482](2001) aglycone ring systemは、1つのbiaryl結合と2つのdiaryl ether結合を含む。 oxyA遺伝子置換mutantから2つのペプチドが分離され、これらはdiaryl ether bridge(C-O-D ring)をひとつ持つだけであった。 --- [PMID: 12404385](2001) oxyAと同様にoxyB, oxyCのmutantを作製し、その産物の構造から各oxygenaseの役割と順番を割り当てている。 1) OxyBによるC-O-D ring systemの形成 2) OxyAによるD-O-E ring systemの形成 3) OxyCによるbiaryl AB systemの形成また、bicyclic C-O-D-O-E ring中間体は、1LeuのN-methylationと、4HpgのO-glycosylationの基質となりえることもわかった。 --- [PMID: 15651041](2005) dpgA mutantではbicyclic hexapeptide、bpsC mutantではmonocyclic hexapeptideの蓄積があることから、OxyBによるCD ring形成はBpsB_module 6からBpsC_module 7へのhexapeptideの移動中かその少し前に起こるはずだと述べている。bpsC mutantでbicyclic hexapeptideが検出されないのは、BpsCでのdeletionのせいでOxyAとNRPS複合体の間の相互作用に必須な部位がないからであると説明されうる。また、dpgA mutantにおいて、7つめのAAとしてDpgの代わりにL-4-hydroxyphenylglycine(Hpg)を含むtricyclic heptapeptideが得られたことから、BpsCとoxygenasesはいくらかの基質寛容性を示す。 --- [PMID: 16730832](2006) oxyA, oxyB, oxyCの非極性mutantsを作製し、閉環反応の順番を再確認している。また、これらmutant株をvancomycin産生菌A. orientalis DSM40040由来の対応するoxygenase genesで補完すると、balhimycin産生は回復した。

Sequence

>cytochrome P450 [P450 monooxygenase]
MFEESNALRGTEIHRRDRFAPGPELRSLMGEGTMSILQPPDSPGGRTGWLATGHDEVRQV
LGSDKFSAKLLYGGTVAGRIWPGFLNQYDPPEHTRLRRMVTSAFTVRRMQDFRPRIEQIV
QASLDAIEAAGGPVDFVPRFAWSVATTVTCDFLGIPRDDQADLSRALHASRSERSGKRRV
AAGNKYWTYMTEIAARARRDPGDDMFGAVVRDHGDAITDAELLGVAAFVMGAGGDQVARF
LAAGAWLMVEHPDQFALLREKPDTVPDWLNEVERYLTSDEKTTPRIAQEDVRIGDQLVKA
GDAVTCSLLAANRRKFPAPEDEFDITRERPVHVTFGHGIHHCLGRPLAEMVFRAAIPALA
QRFPKLRLAEPDREIKLGPPPFDVEALLLEW

>cytochrome P450 [P450 monooxygenase]
GTGTTCGAGGAGAGCAACGCCCTCCGGGGCACGGAAATACACCGGAGAGACCGGTTCGCT
CCGGGGCCGGAACTGCGCTCCCTGATGGGCGAGGGCACCATGTCCATCCTGCAGCCCCCG
GATTCCCCCGGCGGGCGGACCGGGTGGCTGGCCACCGGGCACGACGAGGTCCGGCAGGTC
CTCGGCTCGGACAAGTTCAGCGCCAAGCTGCTCTACGGCGGGACCGTGGCCGGCCGCATC
TGGCCGGGCTTCCTCAACCAGTACGACCCCCCGGAGCACACGCGCCTGCGCCGGATGGTG
ACGTCGGCGTTCACCGTCCGGCGGATGCAGGACTTCCGGCCGCGGATCGAGCAGATCGTC
CAGGCGAGCCTGGACGCCATCGAGGCCGCCGGTGGCCCGGTGGACTTCGTCCCCCGGTTC
GCCTGGTCCGTGGCGACGACGGTGACGTGCGACTTCCTCGGCATCCCGCGTGACGATCAG
GCGGACTTGTCGCGCGCCCTGCACGCCAGCCGGTCCGAACGGTCGGGCAAGCGGCGGGTG
GCGGCGGGGAACAAGTACTGGACGTACATGACCGAGATCGCGGCCCGCGCGCGCCGCGAT
CCCGGTGACGACATGTTCGGCGCGGTGGTGCGCGACCACGGCGACGCGATCACCGACGCG
GAACTGCTGGGCGTGGCCGCGTTCGTCATGGGCGCGGGCGGGGACCAGGTGGCCCGGTTT
CTCGCGGCGGGCGCGTGGCTGATGGTCGAGCACCCCGATCAGTTCGCGCTGCTGCGGGAA
AAGCCGGACACCGTCCCGGACTGGCTGAACGAGGTGGAGCGGTACCTCACCAGCGACGAG
AAGACCACTCCGCGCATCGCGCAGGAGGACGTGCGCATCGGTGATCAGCTCGTCAAGGCC
GGCGATGCCGTCACCTGCTCGCTGCTGGCGGCGAACCGCAGGAAGTTCCCCGCCCCGGAG
GACGAGTTCGACATCACCCGGGAACGGCCGGTGCACGTCACGTTCGGCCACGGCATCCAC
CACTGCCTCGGCAGGCCACTGGCCGAGATGGTGTTCCGGGCGGCGATTCCGGCGCTGGCA
CAACGCTTTCCCAAGCTGAGGCTGGCCGAGCCGGACCGCGAGATCAAGCTGGGGCCGCCG
CCGTTCGACGTGGAAGCCCTGCTGCTGGAGTGGTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR001128 Cytochrome P450 (Family) PF00067 p450 SSF48264 Cytochrome_P450 G3DSA:1.10.630.10 Cyt_P450 IPR002397 Cytochrome P450, B-class (Family) PR00359 BP450 IPR017972 Cytochrome P450, conserved site (Conserved_site) PS00086 CYTOCHROME_P450
SignalP	No significant hit
TMHMM	No significant hit

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