1176834-99-3Relevant academic research and scientific papers
In vitrocharacterization of 3-chloro-4-hydroxybenzoic acid building block formation in ambigol biosynthesis
Kresna, I Dewa Made,Linares-Otoya, Luis,Milzarek, Tobias,Duell, Elke R.,Mir Mohseni, Mahsa,Mettal, Ute,K?nig, Gabriele M.,Gulder, Tobias A. M.,Sch?berle, Till F.
, p. 2302 - 2311 (2021)
The cyanobacteriumFischerella ambiguais a natural producer of polychlorinated aromatic compounds, the ambigols A-E. The biosynthetic gene cluster (BGC) of these highly halogenated triphenyls has been recently identified by heterologous expression. It consists of 10 genes namedab1-10. Two of the encoded enzymes,i.e.Ab2 and Ab3, were identified byin vitroandin vivoassays as cytochrome P450 enzymes responsible for biaryl and biaryl ether formation. The key substrate for these P450 enzymes is 2,4-dichlorophenol, which in turn is derived from the precursor 3-chloro-4-hydroxybenzoic acid. Here, the biosynthetic steps leading towards 3-chloro-4-hydroxybenzoic acid were investigated byin vitroassays. Ab7, an isoenzyme of a 3-deoxy-7-phosphoheptulonate (DAHP) synthase, is involved in chorismate biosynthesis by the shikimate pathway. Chorismate in turn is further converted by a dedicated chorismate lyase (Ab5) yielding 4-hydroxybenzoic acid (4-HBA). The stand alone adenylation domain Ab6 is necessary to activate 4-HBA, which is subsequently tethered to the acyl carrier protein (ACP) Ab8. The Ab8 bound substrate is chlorinated by Ab10 inmetaposition yielding 3-Cl-4-HBA, which is then transfered by the condensation (C) domain to the peptidyl carrier protein and released by the thioesterase (TE) domain of Ab9. The released product is then expected to be the dedicated substrate of the halogenase Ab1 producing the monomeric ambigol building block 2,4-dichlorophenol.
A mechanism-based fluorescence transfer assay for examining ketosynthase selectivity
Prasad, Gitanjeli,Borketey, Lawrence S.,Lin, Tsung-Yi,Schnarr, Nathan A.
experimental part, p. 6717 - 6723 (2012/09/22)
Since their discovery, polyketide synthases have received massive attention from researchers hoping to harness their potential as a platform for generating new and improved therapeutics. Despite significant strides toward this end, inherent specificities within the enzymes responsible for polyketide production have severely limited these efforts. We have developed a mechanism-based, fluorescence transfer assay for a key enzyme component of all polyketide synthases, the ketosynthase domain. As demonstrated, this method can be used with both ketosynthase-containing didomains and full modules. As proof of principle, the ketosynthase domain from module 6 of the 6-deoxyerythronolide synthase is examined for its ability to accept a variety of simple thioester substrates. Consistent with its natural hexaketide substrate, we find that this ketosynthase prefers longer, α-branched thioesters and its ability to distinguish these structural features is quite remarkable. Substrate electronics are also tested via a variety of p-substituted aromatic groups. In all, we expect this technique to find considerable use in the field of polyketide biosynthesis and engineering due to its extraordinary simplicity and very distinct visible readout.
Unnatural polyketide analogues selectively target the her signaling pathway in human breast cancer cells
Kwon, Seok Joon,Kim, Moon Il,Ku, Bosung,Coulombel, Lydie,Kim, Jin-Hwan,Shawky, Joseph H.,Linhardtd, Robert J.,Dordick, Jonathan S.
experimental part, p. 573 - 580 (2011/02/22)
Receptor tyrosine kinases are critical targets for the regulation of cell survival. Cancer patients with abnormal receptor tyrosine kinases (RTK) tend to have more aggressive disease with poor clinical outcomes. As a result, human epidermal growth factor
In vitro precursor-directed synthesis of polyketide analogues with coenzyme a regeneration for the development of antiangiogenic agents
Kim, Moon I.I.,Kwon, Seok Joon,Dordick, Jonathan S.
supporting information; experimental part, p. 3806 - 3809 (2009/12/09)
Polyketide analogues are produced via in vitro reconstruction of a precursor-directed polyketide biosynthetic pathway. Malonyl-CoA synthetase (MCS) was used in conjunction with chalcone synthase (CHS), thereby allowing efficient use of synthetic starter molecules and malonate as extender. Coenzyme-A was recycled up to 50 times. The use of a simple immobilization procedure resulted in up to a 30-fold higher yield of pyrone CHS products than that obtained with the free enzyme solutions.
