Pikro_00170 : CDS information

Location

Organism	Streptomyces venezuelae
Strain	ATCC 15439
Entry name	Pikromycin
Contig	AF079139
Start / Stop / Direction	52,311 / 53,561 / + [in whole cluster] 122 / 1,372 / + [in contig]
Location	52311..53561 [in whole cluster] 122..1372 [in contig]
Type	CDS
Length	1,251 bp (416 aa)

Annotation

Category	3.3 modification reduction
Product	cytochrome P450
Product (GenBank)	cytochrome P450 hydroxylase PiKC
Gene	pikC picK
Gene (GenBank)	pikC
EC number
Keyword	C-12 hydroxylate
Note
Note (GenBank)	cytochrome P450 monooxygenase
Reference	ACC O87605 PmId [9770448] A gene cluster for macrolide antibiotic biosynthesis in Streptomyces venezuelae: architecture of metabolic diversity. (Proc Natl Acad Sci U S A. , 1998) [9778370] Characterization of the macrolide P-450 hydroxylase from Streptomyces venezuelae which converts narbomycin to picromycin. (Biochemistry. , 1998) [9831532] Hydroxylation of macrolactones YC-17 and narbomycin is mediated by the pikC-encoded cytochrome P450 in Streptomyces venezuelae. (Chem Biol. , 1998) [11720530] Isolation and structure determination of novamethymycin, a new bioactive metabolite of the methymycin biosynthetic pathway in Streptomyces venezuelae. (J Nat Prod. , 2001) [16724858] Neopikromycin and novapikromycin from the pikromycin biosynthetic pathway of Streptomyces venezuelae. (J Nat Prod. , 2006) [16825192] The structural basis for substrate anchoring, active site selectivity, and product formation by P450 PikC from Streptomyces venezuelae. (J Biol Chem. , 2006) [17915876] Engineering and analysis of a self-sufficient biosynthetic cytochrome P450 PikC fused to the RhFRED reductase domain. (J Am Chem Soc. , 2007) [19124459] Analysis of transient and catalytic desosamine-binding pockets in cytochrome P-450 PikC from Streptomyces venezuelae. (J Biol Chem. , 2009) [19833867] Selective oxidation of carbolide C-H bonds by an engineered macrolide P450 mono-oxygenase. (Proc Natl Acad Sci U S A. , 2009) [24627965] Directing group-controlled regioselectivity in an enzymatic C-H bond oxygenation. (J Am Chem Soc. , 2014) [26201742] Enzymatic hydroxylation of an unactivated methylene C-H bond guided by molecular dynamics simulations. (Nat Chem. , 2015) comment pikromycin→pikC picromycin→picK pikromycin=picromycin UniProt entryにも両者が登録されている。 --- [PMID: 9770448](1998) 12員環骨格を持つMethymycin and Neomethymycinと、14員環骨格を持つNarbomycin and Pikromycinの生合成で共有されるclusterの同定。 PikC: P450 hydroxylase pikC挿入変異株を作成し、YC-17 (at C-10 and C-12) と narbomycin (at C-12)の両方のhydroxylationを行っていることを確認している。 --- [PMID: 9778370](1998)abstract E.coliで発現させたPicKが、narbomycin → picromycinへのC-12 hydroxylationを担うことを確認。 kinetic analysisあり。 --- [PMID: 9831532](1998) E.coliで発現、精製されたPikCがin vitroで・YC-17のC-10をhydroxylationしてmethymycin ・YC-17のC-12をhydroxylationしてneomethymycin ・narbomycinのC-12をhydroxylationしてpikromycin を産生することを確認。kinetic analysisあり。 --- [PMID: 11720530](2001) abstract PikCを使った生物変換実験で、methymycinのC-12をhydroxylationしてnovamethymycinにすることを確認。 --- [PMID: 16724858](2006) abstract 精製されたPikCはin vitroで・narbomycinのC-14をhydroxylationしてneopikromycinにする・pikromycinのC-14をhydroxylationしてnovapikromycinにする --- [PMID: 16825192](2006) X線構造解析、site-directed mutagenesisに基づくPikCの機能調査。 --- [PMID: 17915876](2007) Rhodococcus sp. NCIMB 9784由来P450RhFはN末側にCYP450を、C末側にreductaseのdomainを持ち、自給自足できるfusionタンパク。PikCを利用しやすくするため、RhFRED domainとのfusionタンパクを作製し、酸化還元反応が自足自給できるようにしている。 --- [PMID: 19124459](2009) [PMID: 16825192]での結果をふまえ、D50N置換でPikC活性が拡大することについてX線構造解析、site-directed mutagenesisなどで調査している。desosamine-binding pocketに関与する。 --- [PMID: 19833867](2009) 著しい基質柔軟性、wild-typeと比べて有意に増加した活性、自給自足性を持つPikC(D50N)-RhFREDが、desosamine glycosideに結合した一連のcarbocyclic環をhydroxylateすることを実証。 --- [PMID: 24627965](2014) abstract 基質のdesosamine部分を置換して、PikCの位置選択性に関する要素を調査している。

Sequence

>cytochrome P450 [cytochrome P450 hydroxylase PiKC]
MRRTQQGTTASPPVLDLGALGQDFAADPYPTYARLRAEGPAHRVRTPEGDEVWLVVGYDR
ARAVLADPRFSKDWRNSTTPLTEAEAALNHNMLESDPPRHTRLRKLVAREFTMRRVELLR
PRVQEIVDGLVDAMLAAPDGRADLMESLAWPLPITVISELLGVPEPDRAAFRVWTDAFVF
PDDPAQAQTAMAEMSGYLSRLIDSKRGQDGEDLLSALVRTSDEDGSRLTSEELLGMAHIL
LVAGHETTVNLIANGMYALLSHPDQLAALRADMTLLDGAVEEMLRYEGPVESATYRFPVE
PVDLDGTVIPAGDTVLVVLADAHRTPERFPDPHRFDIRRDTAGHLAFGHGIHFCIGAPLA
RLEARIAVRALLERCPDLALDVSPGELVWYPNPMIRGLKALPIRWRRGREAGRRTG

>cytochrome P450 [cytochrome P450 hydroxylase PiKC]
GTGCGCCGTACCCAGCAGGGAACGACCGCTTCTCCCCCGGTACTCGACCTCGGGGCCCTG
GGGCAGGATTTCGCGGCCGATCCGTATCCGACGTACGCGAGACTGCGTGCCGAGGGTCCG
GCCCACCGGGTGCGCACCCCCGAGGGGGACGAGGTGTGGCTGGTCGTCGGCTACGACCGG
GCGCGGGCGGTCCTCGCCGATCCCCGGTTCAGCAAGGACTGGCGCAACTCCACGACTCCC
CTGACCGAGGCCGAGGCCGCGCTCAACCACAACATGCTGGAGTCCGACCCGCCGCGGCAC
ACCCGGCTGCGCAAGCTGGTGGCCCGTGAGTTCACCATGCGCCGGGTCGAGTTGCTGCGG
CCCCGGGTCCAGGAGATCGTCGACGGGCTCGTGGACGCCATGCTGGCGGCGCCCGACGGC
CGCGCCGATCTGATGGAGTCCCTGGCCTGGCCGCTGCCGATCACCGTGATCTCCGAACTC
CTCGGCGTGCCCGAGCCGGACCGCGCCGCCTTCCGCGTCTGGACCGACGCCTTCGTCTTC
CCGGACGATCCCGCCCAGGCCCAGACCGCCATGGCCGAGATGAGCGGCTATCTCTCCCGG
CTCATCGACTCCAAGCGCGGGCAGGACGGCGAGGACCTGCTCAGCGCGCTCGTGCGGACC
AGCGACGAGGACGGCTCCCGGCTGACCTCCGAGGAGCTGCTCGGTATGGCCCACATCCTG
CTCGTCGCGGGGCACGAGACCACGGTCAATCTGATCGCCAACGGCATGTACGCGCTGCTC
TCGCACCCCGACCAGCTGGCCGCCCTGCGGGCCGACATGACGCTCTTGGACGGCGCGGTG
GAGGAGATGTTGCGCTACGAGGGCCCGGTGGAATCCGCGACCTACCGCTTCCCGGTCGAG
CCCGTCGACCTGGACGGCACGGTCATCCCGGCCGGTGACACGGTCCTCGTCGTCCTGGCC
GACGCCCACCGCACCCCCGAGCGCTTCCCGGACCCGCACCGCTTCGACATCCGCCGGGAC
ACCGCCGGCCATCTCGCCTTCGGCCACGGCATCCACTTCTGCATCGGCGCCCCCTTGGCC
CGGTTGGAGGCCCGGATCGCCGTCCGCGCCCTTCTCGAACGCTGCCCGGACCTCGCCCTG
GACGTCTCCCCCGGCGAACTCGTGTGGTATCCGAACCCGATGATTCGCGGGCTCAAGGCC
CTGCCGATCCGCTGGCGGCGAGGACGGGAGGCGGGCCGCCGTACCGGTTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR001128 Cytochrome P450 (Family) PR00385 P450 PF00067 p450 G3DSA:1.10.630.10 Cyt_P450 SSF48264 Cytochrome_P450 IPR002397 Cytochrome P450, B-class (Family) PR00359 BP450 IPR017972 Cytochrome P450, conserved site (Conserved_site) PS00086 CYTOCHROME_P450
SignalP	Bacteria, Gram-negative
TMHMM	No significant hit

Pikro_00160 Pikro_00180