Asco_00150 : CDS information

Location

Organism	Streptomyces hygroscopicus subsp. ascomyceticus
Strain	ATCC 14891 (=NBRC 13981)
Entry name	Ascomycin
Contig	AF235504
Start / Stop / Direction	46,752 / 47,786 / + [in whole cluster] 46,752 / 47,786 / + [in contig]
Location	46752..47786 [in whole cluster] 46752..47786 [in contig]
Type	CDS
Length	1,035 bp (344 aa)

Annotation

Category	2.2 modification addition of starter unit
Product	chorismatase chorismate hydrolase
Product (GenBank)	FkbO
Gene
Gene (GenBank)	fkbO
EC number
Keyword	4,5-dihydroxycyclohex-1-ene carboxylic acid
Note
Note (GenBank)	9-hydroxyl oxidase
Reference	ACC Q9KID9 PmId [10863099] The FK520 gene cluster of Streptomyces hygroscopicus var. ascomyceticus (ATCC 14891) contains genes for biosynthesis of unusual polyketide extender units. (Gene. , 2000) [21383123] Biosynthesis of the immunosuppressants FK506, FK520, and rapamycin involves a previously undescribed family of enzymes acting on chorismate. (Proc Natl Acad Sci U S A. , 2011) [24036425] Mechanistic implications for the chorismatase FkbO based on the crystal structure. (J Mol Biol. , 2014) [26247872] Chorismatase Mechanisms Reveal Fundamentally Different Types of Reaction in a Single Conserved Protein Fold. (J Am Chem Soc. , 2015) comment [PMID: 10863099](2000) FK520 gene clusterの報告。 fkbO: C9 hydroxyl oxidase PKSによって合成された中間体はFkbPによって縮合されてPKS complexから放出されると、alpha-ketoamide functionを得るためにC9がFkbDによってhydroxylateされ、FkbOによって酸化される。実験的裏づけはなし。 -- [PMID: 21383123](2011) FK520 gene clusterのfkbOをE.coli発現、精製。このタンパクでChorismateは(4R,5R)-4,5-dihydroxycyclohexa-1,5-dienecarboxylic acid(=DCDC)へ変換されたことを、産物解析により確認。 kinetic parametersは反応産物pyruvateをモニターして測定。 chorismate + H2O → DCDC + pyruvate rapamycin生合成において、(4R,5R)-4,5-dihydroxycyclohex-1-enecarboxylic acid(=DHCHC)がstarter unit前駆体であり、その産生にはrapKが必須なことがわかっている。 rapK deletion mutantはrapamycin非産生。 FK506 or FK520 gene cluster由来fkbOで補完され、rapamycin産生回復。 --- [PMID:24036425](2013) 結晶構造解析から、FkbOの反応モデルを提唱。 active-site変異体の活性測定でそれを確認している。

Sequence

>chorismatase [FkbO]
MTDAGRQGRVEALSISVTAPYCRFEKTGSPDLEGDETVLGLIEHGTGHTDVSLVDGAPRT
AVHTTTRDDEAFTEVWHAQRPVESGMDNGIAWARTDAYLFGVVRTGESGRYADATAALYT
NVFQLTRSLGYPLLARTWNYVSGINTTNADGLEVYRDFCVGRAQALDEGGIDPATMPAAT
GIGAHGGGITCVFLAARGGVRINIENPAVLTAHHYPTTYGPRPPVFARATWLGPPEGGRL
FISATAGILGHRTVHHGDVTGQCEVALDNMARVIGAENLRRHGVQRGHVLADVDHLKVYV
RRREDLDTVRRVCAARLSSTAAVALLHTDIAREDLLVEIEGMVA

>chorismatase [FkbO]
ATGACCGATGCCGGACGCCAGGGAAGGGTGGAGGCGTTGTCCATATCTGTCACGGCGCCG
TATTGCCGCTTCGAGAAGACCGGATCACCGGACCTCGAGGGTGACGAGACGGTGCTCGGC
CTGATCGAGCACGGCACCGGCCACACCGACGTGTCGCTGGTGGACGGTGCTCCCCGGACC
GCCGTGCACACCACGACCCGTGACGACGAGGCGTTCACCGAGGTCTGGCACGCACAGCGC
CCTGTCGAGTCCGGCATGGACAACGGCATCGCCTGGGCCCGCACCGACGCGTACCTGTTC
GGTGTCGTGCGCACCGGCGAGAGCGGCAGGTACGCCGATGCCACCGCGGCCCTCTACACG
AACGTCTTCCAGCTCACCCGGTCGCTGGGGTATCCCCTGCTCGCCCGGACCTGGAACTAC
GTCAGCGGTATCAACACGACGAACGCGGACGGGCTGGAGGTGTACCGGGACTTCTGCGTG
GGCCGCGCCCAGGCGCTCGACGAGGGCGGGATCGACCCGGCCACCATGCCCGCGGCCACC
GGTATCGGCGCCCACGGGGGCGGCATCACCTGCGTGTTCCTCGCCGCCCGGGGCGGAGTG
CGGATCAACATCGAGAACCCCGCCGTCCTCACGGCCCACCACTACCCGACGACGTACGGT
CCGCGGCCCCCGGTCTTCGCACGGGCCACCTGGCTGGGCCCGCCGGAGGGGGGCCGGCTG
TTCATCTCCGCGACGGCCGGCATCCTCGGACACCGAACGGTGCACCACGGTGATGTGACC
GGCCAGTGCGAGGTCGCCCTCGACAACATGGCCCGGGTCATCGGCGCGGAGAACCTGCGG
CGCCACGGCGTCCAGCGGGGGCACGTCCTCGCCGACGTGGACCACCTCAAGGTCTACGTC
CGCCGCCGCGAGGATCTCGATACGGTCCGCCGGGTCTGCGCCGCACGCCTGTCGAGCACC
GCGGCCGTCGCCCTTTTGCACACCGACATAGCCCGCGAGGATCTGCTCGTCGAAATCGAA
GGCATGGTGGCGTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR013813 Endoribonuclease L-PSP/chorismate mutase-like (Domain) G3DSA:3.30.1330.40 Endoribo_LPSP/chorism_mut-like SSF55298 YjgF/chorismate_mutase-like
SignalP	No significant hit
TMHMM	No significant hit

Asco_00140 Asco_00160