- RadH: A Versatile Halogenase for Integration into Synthetic Pathways
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Flavin-dependent halogenases are useful enzymes for providing halogenated molecules with improved biological activity, or intermediates for synthetic derivatization. We demonstrate how the fungal halogenase RadH can be used to regioselectively halogenate a range of bioactive aromatic scaffolds. Site-directed mutagenesis of RadH was used to identify catalytic residues and provide insight into the mechanism of fungal halogenases. A high-throughput fluorescence screen was also developed, which enabled a RadH mutant to be evolved with improved properties. Finally we demonstrate how biosynthetic genes from fungi, bacteria, and plants can be combined to encode a new pathway to generate a novel chlorinated coumarin “non-natural” product in E. coli.
- Menon, Binuraj R. K.,Brandenburger, Eileen,Sharif, Humera H.,Klemstein, Ulrike,Shepherd, Sarah A.,Greaney, Michael F.,Micklefield, Jason
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supporting information
p. 11841 - 11845
(2017/09/20)
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- Specific chlorination of isoquinolines by a fungal flavin-dependent halogenase
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Rdc2 is the first flavin-dependent halogenase identified from fungi. Based on the reported structure of the bacterial halogenase CmlS, we have built a homology model for Rdc2. The model suggests an open substrate binding site that is capable of binding the natural substrate, monocillin II, and possibly other molecules such as 4-hydroxyisoquinoline (1) and 6-hydroxyisoquinoline (2). In vitro and in vivo halogenation experiments confirmed that 1 and 2 can be halogenated at the position ortho to the hydroxyl group, leading to the synthesis of the chlorinated isoquinolines 1a and 2a, respectively, which further expands the spectrum of identified substrates of Rdc2. This work revealed that Rdc2 is a useful biocatalyst for the synthesis of various halogenated compounds.
- Zeng, Jia,Lytle, Anna K.,Gage, David,Johnson, Sean J.,Zhan, Jixun
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p. 1001 - 1003
(2013/03/13)
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- CYCLOALKYLAMINE SUBSTITUTED ISOQUINOLINE DERIVATIVES
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The invention relates to 6-substituted isoquinoline derivatives of the Formula (I) useful for the treatment and/or prevention of diseases associated with Rho-kinase and/or Rho-kinase mediated phosphorylation of myosin light chain phosphatase, and compositions containing such compounds.
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Page/Page column 44
(2008/12/06)
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- ISOQUINOLINE DERIVATIVES AS INHIBITORS OF RHO-KINASE
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The invention relates to 6-piperidinyl-substituted isoquinoline derivatives of the formula (I); useful for the treatment and/or prevention of diseases associated with Rho-kinase and/or Rho-kinase mediated phosphorylation of myosin light chain phosphatase, and compositions containing such compounds.
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Page/Page column 35-36
(2008/06/13)
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