1. Volume 20, Issue 4, Pages 510-520 (April 2013)
Biosynthetic Conclusions from the Functional Dissection of Oxygenases for Biosynthesis of Actinorhodin and Related Streptomyces Antibiotics 
Takaaki Taguchi, Masaki Yabe, Hitomi Odaki, Miki Shinozaki, Mikko Metsä-Ketelä, Takao Arai, Susumu Okamoto, Koji Ichinose 
Chemistry & Biology 
Volume 20, Issue 4, Pages (April 2013)
DOI: /j.chembiol
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2. Chemistry & Biology 2013 20, 510-520DOI: (10. 1016/j. chembiol. 2013
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3. Figure 1 Overview of Actinorhodin Biosynthesis
(A) Currently proposed biosynthetic pathway of ACT. The numberings are based on the biosynthetic order. ARO, aromatase; CYC, cyclase. Dashed arrows indicate the shunt pathways.
(B) The structures of BIQ antibiotics, ALN, and ALN-related compounds. DMAC, 3,8-dihydroxy-1-methylanthraquinone-2-carboxylic acid; (S)-DNPA, 4-dihydro-9-hydroxy-1-methyl-10-oxo-3-H-naphtho-[2,3-c]-pyran-3-(S)-acetic acid.
Chemistry & Biology  , DOI: ( /j.chembiol )
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4. Figure 2 In Vitro Quinone-Forming Activities of the Recombinant FMOs
(A) SDS-PAGE analysis of the recombinant FMOs used for in vitro assay.
(B) Quinone-forming activities of the FMOs. Emodinanthrone was used as the substrate. (a) boiled ActVA-ORF5/ActVB as negative control. A small amount of emodin was spontaneously formed by autoxidation, (b) ActVA-ORF5/ActVB, (c) Gra-21/ActVB, (d) Med-7/ActVB, (e) AlnT/ActVB, and (f) AlnT/AlnH. Chromatograms monitored by absorbance at 254 nm are shown.
See also Figure S1 and Table S3.
Chemistry & Biology  , DOI: ( /j.chembiol )
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5. Figure 3 In Vivo Functional Analysis of ActVA-ORF5 and Relevant FMOs
(A) The reverse sides of R5− medium plates. Except for the wild-type strain, all the others are in the ΔactVA-5,6 mutant background.
(B) HPLC chromatograms of the ACT-related metabolites from R5MS liquid culture are shown. Chromatograms were monitored by absorbance at 450 nm. (a) ΔactVA-5,6 harboring pTYM1ep_His as negative control, (b) ΔactVA-5,6/alnT, (c) ΔactVA-5,6/med-ORF7, (d) ΔactVA-5,6/gra-ORF21, (e) ΔactVA-5,6/actVA-ORF5, and (f) M510 harboring pTYM1ep_His as positive control. Under our experimental conditions, ACPL 8 was eluted at 22.2 min, whereas the lactonized derivatives of ACT 1, γ-actinorhodin (γ-ACT, 11) and γ′-actinorhodin (γ′-ACT) were eluted at 23.7 and 22.9 min, respectively. In (d), an enlarged chromatogram depicting a peak of 11 is also shown.
See also Table S3.
Chemistry & Biology  , DOI: ( /j.chembiol )
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6. Figure 4
Production of DHK-OH by Coexpression of the Minimal Gene Set for DDHK Biosynthesis and the FMOs under Study
HPLC analyses of metabolites produced by S. coelicolor CH999/pIJ5659 with (a) gra-ORF21, (b) actVA-ORF5, (c) med-ORF7, and (d) vector control. Chromatograms monitored by absorbance at 450 nm are shown. See also Figure S4.
Chemistry & Biology  , DOI: ( /j.chembiol )
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7. Figure 5 Structures of Compounds 12, 13, and 15
(A) The elucidated structure of compound 12 as 4R-(N-acetyl-cysteinyl)-dihydrokalafungin (4R-AcCys-DHK).
(B) The deduced structure of compound 13 from compound 15, 4R-(N-acetyl-cysteinyl)-kalafungin (4R-AcCys-KAL).
(C) A proposed mechanism of AcCys moiety addition to DHK2 recruiting mycothiol17.
See also Figures S2 and S3; Tables S1 and S2.
Chemistry & Biology  , DOI: ( /j.chembiol )
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8. Figure 6
Proposed Later Tailoring Steps in the Biosyntheses of ACT, GRA, and MED
Chemistry & Biology  , DOI: ( /j.chembiol )
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