Characterization of JadH as an FAD‐ and NAD(P)H‐Dependent Bifunctional Hydroxylase/Dehydrase in Jadomycin Biosynthesis

Chen, Yihua; Fan, Keqiang; He, Yong‐Xing; Xu, Xinping; Peng, Yonghan; Yu, Tingting; Jia, Cui-Juan; Yang, Keqian

doi:10.1002/cbic.201000178

Cited by 32 publications

(41 citation statements)

References 33 publications

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“…Identical findings and conclusions were reported previously for JadH, the homologous enzyme in the jadomycin biosynthesis pathway. [16, 17] Contrary to our previous findings in in vivo experiments, an enzyme mixture that consists of the enzymes GilOI, GilOII, GilM, GilMT, GilR, and Fre [15] was able to completely convert 16 into 1 (see Table S4 in the Supporting Information). This result confirms that 16 is a true intermediate in the gilvocarcin biosynthesis pathway (Figure 3, trace D).…”

contrasting

confidence: 66%

Enzymatic Total Synthesis of Defucogilvocarcin M and Its Implications for Gilvocarcin Biosynthesis

et al. 2011

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contrasting

confidence: 66%

Enzymatic Total Synthesis of Defucogilvocarcin M and Its Implications for Gilvocarcin Biosynthesis

et al. 2011

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“…These studies identified GilOI as the enzyme responsible for the next reaction, the oxidation of 12 to dehydrorabelomycin ( 21 ), proving that GilOI is a bifunctional enzyme which performs a 4a,12b-dehydration and C-12 oxygenation. Identical observations and conclusions were reported previously for JadH, the homologous enzyme of the jadomycin pathway [86,91,92], and now also for LanE (see above).…”

Section: Introductionsupporting

confidence: 88%

Delineation of gilvocarcin, jadomycin, and landomycin pathways through combinatorial biosynthetic enzymology

Kharel

Rohr

2012

Current Opinion in Chemical Biology

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“…Purified AptC and AdaC were thermally denatured and the prosthetic groups were confirmed to be FAD by LC-MS analysis as previously described (Figure S12). 32 To generate malonyl-CoA in situ for use in polyketide turnover assays, we used the malonyl-CoA synthetase MatB from Rhizobium trifolii to produce the extender units from malonate, ATP and CoA. 28 …”

Section: Resultsmentioning

confidence: 99%

Comparative Characterization of Fungal Anthracenone and Naphthacenedione Biosynthetic Pathways Reveals an α-Hydroxylation-Dependent Claisen-like Cyclization Catalyzed by a Dimanganese Thioesterase

et al. 2011

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The linear tetracyclic TAN-1612 (1) and BMS-192548 (2) were isolated from different filamentous fungal strains, and have been examined as potential neuropeptide Y and neurokinin-1 receptor antagonist respectively. Although the biosynthesis of fungal aromatic polyketides has attracted much interest in recent years, the biosynthetic mechanism for such naphthacenedione-containing products has not been established. Using a targeted genome mining approach, we first located the ada gene cluster responsible for the biosynthesis of 1 in Aspergillus niger ATCC 1015. The connection between 1 and the ada pathway was verified through overexpression of the Zn2Cys6-type pathway-specific transcriptional regulator AdaR and subsequent gene expression analysis. The enzymes encoded in the ada gene cluster share high sequence similarities to the known apt pathway linked to the biosynthesis of anthraquinone asperthecin 3. Subsequent comparative investigation of these two highly homologous gene clusters by heterologous pathway reconstitution in Saccharomyces cerevisiae revealed a novel α-hydroxylation-dependent Claisen cyclization cascade, which involves a flavin-dependent monooxygenase (FMO) that hydroxylates the α-carbon of an ACP-bound polyketide and a bifunctional metallo-β-lactamase-type thioesterase (MβL-TE). The bifunctional MβL-TE catalyzes the fourth ring cyclization to afford the naphthacenedione scaffold upon α-hydroxylation, while performs hydrolytic release of an anthracenone product in the absence of α-hydroxylation. Through in vitro biochemical characterizations and metal analyses, we verified that the apt MβL-TE is a dimanganese enzyme and requires both Mn2+ cations for the observed activities. The MβL-TE is the first example of TE in polyketide biosynthesis that catalyzes the Claisen-like condensation without an α/β hydrolase fold and forms no covalent bond with the substrate. These mechanistic features should be general to the biosynthesis of tetracyclic naphthacenedione compounds in fungi.

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Characterization of JadH as an FAD‐ and NAD(P)H‐Dependent Bifunctional Hydroxylase/Dehydrase in Jadomycin Biosynthesis

Cited by 32 publications

References 33 publications

Enzymatic Total Synthesis of Defucogilvocarcin M and Its Implications for Gilvocarcin Biosynthesis

Enzymatic Total Synthesis of Defucogilvocarcin M and Its Implications for Gilvocarcin Biosynthesis

Delineation of gilvocarcin, jadomycin, and landomycin pathways through combinatorial biosynthetic enzymology

Comparative Characterization of Fungal Anthracenone and Naphthacenedione Biosynthetic Pathways Reveals an α-Hydroxylation-Dependent Claisen-like Cyclization Catalyzed by a Dimanganese Thioesterase

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