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Biosynthetic chlorination of the piperazate residue in kutzneride biosynthesis by KthP.
- Source :
-
Biochemistry [Biochemistry] 2011 Jul 12; Vol. 50 (27), pp. 6063-72. Date of Electronic Publication: 2011 Jun 20. - Publication Year :
- 2011
-
Abstract
- Kutznerides 2 and 8 of the cyclic hexadepsipeptide family of antifungal natural products from the soil actinomycete Kutzneria sp. 744 contain two sets of chlorinated residues, a 6,7-dichlorohexahydropyrroloindole moiety derived from dichlorotryptophan and a 5-chloropiperazate moiety, as well as a methylcyclopropylglycine residue that may arise from isoleucine via a cryptic chlorination pathway. Previous studies identified KtzD, KtzQ, and KtzR as three halogenases in the kutzneride pathway but left no candidate for installing the C5 chlorine on piperazate. On the basis of analysis of the complete genome sequence of Kutzneria, we now identify a fourth halogenase in the pathway whose gene is separated from the defined kutzneride cluster by 12 open reading frames. KthP (kutzneride halogenase for piperazate) is a mononuclear nonheme iron halogenase that acts on the piperazyl ring tethered by a thioester linkage to the holo forms of thiolation domains. MS analysis of the protein-bound product confirmed chlorination of the piperazate framework from the (3S)- but not the (3R)-piperazyl-S-pantetheinyl thiolation proteins. After thioesterase-mediated release, nuclear magnetic resonance was used to assign the free imino acid as (3S,5S)-5-chloropiperazate, distinct from the 3S,5R stereoisomer reported in the mature kutznerides. These results demonstrate that a fourth halogenase, KthP, is active in the kutzneride biosynthetic pathway and suggest further processing of the (3S,5S)-5-chloropiperazate during subsequent incorporation into the kutzneride depsipeptide frameworks.
- Subjects :
- Actinomycetales genetics
Actinomycetales metabolism
Antifungal Agents chemical synthesis
Antifungal Agents metabolism
Depsipeptides genetics
Indoles metabolism
Isoleucine chemistry
Multigene Family
Piperazine
Piperazines metabolism
Proline chemistry
Pyrroles metabolism
Soil Microbiology
Sulfhydryl Compounds chemical synthesis
Tryptophan analogs & derivatives
Tryptophan chemical synthesis
Tryptophan chemistry
Actinomycetales chemistry
Depsipeptides chemical synthesis
Depsipeptides metabolism
Halogenation genetics
Indoles chemical synthesis
Piperazines chemical synthesis
Pyrroles chemical synthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1520-4995
- Volume :
- 50
- Issue :
- 27
- Database :
- MEDLINE
- Journal :
- Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 21648411
- Full Text :
- https://doi.org/10.1021/bi200656k