Rifam_00300 Rifam_00300   Rifam_00320 Rifam_00320

Rifam_00310 : CDS information

close this sectionLocation

Organism
StrainS699
Entry nameRifamycin
Contig
Start / Stop / Direction72,331 / 73,497 / + [in whole cluster]
72,331 / 73,497 / + [in contig]
Location72331..73497 [in whole cluster]
72331..73497 [in contig]
TypeCDS
Length1,167 bp (388 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category2.2 modification addition of starter unit
Product3-amino-5-hydroxybenzoic acid synthase/UDP-3-keto-D-glucose aminotransferase
AHBA synthase/UDP-3-keto-D-glucose aminotransferase
Product (GenBank)RifK
Gene
Gene (GenBank)rifK
EC number4.2.1.144
2.6.1.-
Keyword
  • AHBA
Note
  • bifunctional protein
Note (GenBank)
  • AHBA synthase
Reference
ACC
PmId
[9497318] 3-Amino-5-hydroxybenzoic acid synthase, the terminal enzyme in the formation of the precursor of mC7N units in rifamycin and related antibiotics. (J Biol Chem. , 1998)
[9512878] Biosynthesis of the ansamycin antibiotic rifamycin: deductions from the molecular analysis of the rif biosynthetic gene cluster of Amycolatopsis mediterranei S699. (Chem Biol. , 1998)
[10433690] Crystal structure of 3-amino-5-hydroxybenzoic acid (AHBA) synthase. (Biochemistry. , 1999)
[11278540] Mutational analysis and reconstituted expression of the biosynthetic genes involved in the formation of 3-amino-5-hydroxybenzoic acid, the starter unit of rifamycin biosynthesis in amycolatopsis Mediterranei S699. (J Biol Chem. , 2001)
[12207505] Characterization of the early stage aminoshikimate pathway in the formation of 3-amino-5-hydroxybenzoic acid: the RifN protein specifically converts kanosamine into kanosamine 6-phosphate. (J Am Chem Soc. , 2002)
[21081954] The biosynthesis of 3-amino-5-hydroxybenzoic acid (AHBA), the precursor of mC7N units in ansamycin and mitomycin antibiotics: a review. (J Antibiot (Tokyo). , 2011)
comment
[PMID: 9497318](1998)
S699株使用。
gene name表記なし。AHBA synthase配列記載あり、このORFに一致。
また、rifamycin cluster報告論文[PMID: 9512878]でも引用されている。

AHBA synthaseをA.mediterraneiから精製、クローニング、シークエンス、E.coliで過剰発現、recombinant enzyme活性確認。
AHBA synthaseをコードする遺伝子の不活化でrifamycin非産生、AHBA synthaseで補うと産生回復。

AHBA synthaseは、rifamycin PKSのstarter unitであるAHBAの合成過程で、aminoDHS→AHBAを触媒する。

AHBA: 3-amino-5-hydroxybenzoic acid
aminoDHS: 5-deoxy-5-amino-3-dehydroshikimate

---
[PMID: 9512878](1998)
ansamycin系抗生物質rifamycinの生合成gene clusterの報告。
RifK(388aa): AHBA synthase

上記報告をふまえて、rifK geneを中心にclusterを同定。

---
[PMID: 10433690](1999)
構造解析
active siteは2つのsubunitの残基で編成されるので、AHBA synthaseはhomodimerで活性がある。

---
[PMID: 11278540](2001)
rifamycinと関連ansamycinsのPKS starter 3-amino-5-hydroxybenzoic acid (AHBA)の生合成経路について調査。関連遺伝子の不活化、異種性発現、中間体を基質とした反応確認など。
RifKについては引用で、主にそれ以外のgenesについての検討。

---
[PMID: 12207505](2002)
rif gene cluster内に他に候補がないので、RifKがAHBA synthaseに加えて2つめの機能として、aminoDAHPの前駆体にN原子を導入することが考えられる。
UDP-3-ketoglucose→UDP-kanosamine

aminoDAHP = 4-amino-3,4-dideoxy-D-arabino-heptulosonic acid 7-phosphate

---
[PMID: 21081954](2011)
AHBA生合成に関するreview.
A. mediterraneiにおけるAHBA形成のcomplete pathwayが示されている(Fig. 9)。

発現、精製したoxidoreductase候補RifLに活性が見られないことから、RifLが不安定であり、2つめの機能として酸化されたUDP-glucoseにNを動員するとされるRifKとcomplexを形成して安定化すると考えられる。

rifKLの同時発現で得られたタンパクは、UDP-glucoseでNAD+→NADHする活性が見られたが、dTDP-glucoseでは見られない。
この反応にglutamineを加えると活性は拡大する。よってglutamineがN源である。

RifL: UDP-D-glucose + NAD+ → UDP-3-keto-D-glucose + NADH
RifK: UDP-3-keto-D-glucose + glutamine → UDP-kanosamine

close this sectionSequence

selected fasta
>3-amino-5-hydroxybenzoic acid synthase/UDP-3-keto-D-glucose aminotransferase [RifK]
MNARKAPEFPAWPQYDDAERNGLVRALEQGQWWRMGGDEVNSFEREFAAHHGAAHALAVT
NGTHALELALQVMGVGPGTEVIVPAFTFISSSQAAQRLGAVTVPVDVDAATYNLDPEAVA
AAVTPRTKVIMPVHMAGLMADMDALAKISADTGVPLLQDAAHAHGARWQGKRVGELDSIA
TFSFQNGKLMTAGEGGAVVFPDGETEKYETAFLRHSCGRPRDDRRYFHKIAGSNMRLNEF
SASVLRAQLARLDEQIAVRDEPWTLLSRLLGAIDGVVPQGGDVRADRNSHYMAMFRIPGL
TEERRNALVDRLVEAGLPAFAAFRAIYRTDAFWELGAPDESVDAIARRCPNTDAISSDCV
WLHHRVLLAGEPELHATAEIIADAVGRA
selected fasta
>3-amino-5-hydroxybenzoic acid synthase/UDP-3-keto-D-glucose aminotransferase [RifK]
ATGAACGCGCGAAAGGCACCGGAATTCCCTGCGTGGCCGCAGTACGACGACGCCGAGCGG
AACGGCTTGGTCCGCGCGCTCGAACAGGGTCAGTGGTGGCGCATGGGCGGGGACGAGGTG
AACTCCTTCGAGCGCGAGTTCGCCGCCCACCACGGCGCCGCGCACGCGCTGGCGGTCACC
AACGGCACGCACGCGCTGGAACTCGCCCTGCAGGTCATGGGCGTCGGCCCGGGCACCGAG
GTCATCGTGCCGGCCTTCACCTTCATCTCCTCGTCCCAGGCGGCCCAGCGGCTCGGCGCG
GTCACCGTCCCGGTCGACGTCGACGCGGCCACCTACAACCTCGACCCGGAGGCCGTCGCG
GCCGCCGTCACCCCGCGCACCAAGGTGATCATGCCGGTGCACATGGCCGGGCTGATGGCC
GACATGGACGCGCTGGCGAAGATCTCCGCCGACACGGGCGTCCCGCTGCTGCAGGACGCC
GCGCACGCGCACGGCGCCCGCTGGCAGGGCAAGCGCGTCGGCGAGCTGGACAGCATCGCC
ACGTTCAGCTTCCAGAACGGCAAGCTGATGACGGCGGGCGAGGGCGGCGCGGTCGTCTTC
CCCGACGGCGAGACCGAAAAGTACGAGACCGCCTTCCTCCGGCACAGCTGCGGCCGTCCC
CGCGACGACCGCCGCTACTTCCACAAGATCGCCGGCTCGAACATGCGGCTCAACGAGTTC
TCCGCGTCCGTGCTGCGCGCGCAGCTGGCCCGCCTCGACGAGCAGATCGCCGTGCGCGAC
GAGCCGTGGACCCTGCTGTCCCGGCTGCTCGGCGCGATCGACGGCGTCGTGCCCCAGGGC
GGCGACGTGCGCGCCGACCGCAACTCCCACTACATGGCCATGTTCCGCATCCCCGGGCTC
ACCGAGGAACGGCGCAACGCCCTGGTCGACCGGCTCGTCGAGGCCGGCCTGCCCGCCTTC
GCCGCGTTCCGCGCGATCTACCGCACCGACGCCTTCTGGGAGCTCGGCGCCCCCGACGAG
AGCGTGGACGCGATCGCCCGGCGCTGCCCGAACACGGACGCGATCAGCAGCGACTGCGTC
TGGCTGCACCACCGGGTCCTGCTGGCCGGCGAGCCGGAGCTGCACGCGACCGCCGAAATC
ATCGCCGACGCCGTGGGCCGGGCATGA

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR000653 DegT/DnrJ/EryC1/StrS aminotransferase (Family)
[1-388] 9.49996855930963e-110 PIRSF000390
PIRSF000390   PLP_StrS
[15-372] 1.9e-117 PF01041
PF01041   DegT_DnrJ_EryC1
IPR015421 Pyridoxal phosphate-dependent transferase, major region, subdomain 1 (Domain)
[11-250] 4.09999999999995e-73 G3DSA:3.40.640.10
G3DSA:3.40.640.10   PyrdxlP-dep_Trfase_major_sub1
IPR015422 Pyridoxal phosphate-dependent transferase, major region, subdomain 2 (Domain)
[251-386] 1e-60 G3DSA:3.90.1150.10
G3DSA:3.90.1150.10   PyrdxlP-dep_Trfase_major_sub2
IPR015424 Pyridoxal phosphate-dependent transferase, major domain (Domain)
[5-388] 7.60000011115808e-101 SSF53383
SSF53383   PyrdxlP-dep_Trfase_major
SignalP No significant hit
TMHMM No significant hit
Rifam_00300 Rifam_00300   Rifam_00320 Rifam_00320
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