First in vitro directed biosynthesis of new compounds by a minimal type II polyketide synthase: evidence for the mechanism of chain length determinationElectronic supplementary information (ESI) available: LCMS spectra and detailed experimental procedures for all reported experiments. See http://www.rsc.org/suppdata/cc/b3/b300847a/

2003 ◽  
pp. 686-687 ◽  
Author(s):  
T. P. Nicholson ◽  
C. Winfield ◽  
J. Westcott ◽  
J. Crosby ◽  
T. J. Simpson ◽  
...  
Keyword(s):  
Chain Length ◽  
Polyketide Synthase ◽  
Supplementary Information ◽  
Type Ii ◽  
Directed Biosynthesis ◽  
Minimal Type ◽  
New Compounds

scholarly journals Evaluating precursor-directed biosynthesis towards novel erythromycins through in vitro studies on a bimodular polyketide synthase

1998 ◽  
Vol 5 (12) ◽  
pp. 743-754 ◽  
Author(s):  
Kira J. Weissman ◽  
Matthew Bycroft ◽  
Annabel L. Cutter ◽  
Ulf Hanefeld ◽  
Elizabeth J. Frost ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
In Vitro Studies ◽  
Directed Biosynthesis

scholarly journals Aromatic polyketide biosynthesis: fidelity, evolution and engineering

2019 ◽  
Author(s):  
Zhiwei Qin ◽  
Rebecca Devine ◽  
Matthew I. Hutchings ◽  
Barrie Wilkinson
Keyword(s):  
Biosynthetic Pathway ◽  
Polyketide Synthase ◽  
Single Gene ◽  
Biosynthetic Gene Cluster ◽  
Type Ii ◽  
Trace Level ◽  
Aromatic Polyketides ◽  
Pyridine Moiety ◽  
Catalytic Role ◽  
New Compounds

AbstractWe report the formicapyridines which are structurally and biosynthetically related to the pentacyclic fasamycin and formicamycin aromatic polyketides but comprise a rare pyridine moiety. These new compounds are trace level metabolites formed by derailment of the major biosynthetic pathway. Inspired by evolutionary logic we show that rational mutation of a single gene in the biosynthetic gene cluster leads to a significant increase both in total formicapyridine production and their enrichment relative to the fasamycins/formicamycins. Our observations broaden the polyketide biosynthetic landscape and identify a non-catalytic role for ABM superfamily proteins in type II polyketide synthase assemblages for maintaining biosynthetic pathway fidelity.

scholarly journals Widening the bottleneck: Heterologous expression, purification, and characterization of the Ktedonobacter racemifer minimal type II polyketide synthase in Escherichia coli

2020 ◽  
Author(s):  
Joshua G. Klein ◽  
Yang Wu ◽  
Bashkim Kokona ◽  
Louise K. Charkoudian
Keyword(s):  
Heterologous Expression ◽  
Polyketide Synthase ◽  
Analytical Ultracentrifugation ◽  
Acyl Carrier Protein ◽  
Unmet Need ◽  
Type Ii ◽  
E Coli ◽  
Potential Impact

AbstractEnzyme assemblies such as type II polyketide synthases (PKSs) produce a wide array of bioactive secondary metabolites. While the molecules produced by type II PKSs have found remarkable success in the clinic, the biosynthetic prowess of these enzymes has been stymied by: 1) the inability to reconstitute the bioactivity of the minimal PKS enzymes in vitro and 2) limited exploration of type II PKSs from diverse phyla. Towards filling this unmet need, we expressed, purified, and characterized the ketosynthase chain length factor (KSCLF) and acyl carrier protein (ACP) from Ktedonobacter racemifer. Using E. coli as a heterologous host, we obtained soluble proteins in titers representing significant improvements over previous KSCLF heterologous expression efforts. Characterization of these enzymes reveals that KrACP has self-malonylating activity. Sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis of holo-KrACP and KrKSCLF indicates that these enzymes do not interact in vitro, suggesting that the acylated state of these proteins might play an important role in facilitating biosynthetically relevant interactions. Given the potential impact of obtaining soluble core type II PKS biosynthetic enzymes to enable in vitro characterization studies, these results lay important groundwork for optimizing the interaction between KrKSCLF and KrACP and exploring the biosynthetic potential of other non-actinomycete type II PKSs.

scholarly journals Comparison of the Antiviral Activities of Alkoxyalkyl and Alkyl Esters of Cidofovir against Human and Murine Cytomegalovirus Replication In Vitro

2005 ◽  
Vol 49 (2) ◽  
pp. 656-662 ◽  
Author(s):  
William B. Wan ◽  
James R. Beadle ◽  
Caroll Hartline ◽  
Earl R. Kern ◽  
Stephanie L. Ciesla ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Chain Length ◽  
Murine Cytomegalovirus ◽  
Alkyl Esters ◽  
Highly Active ◽  
Antiviral Activities ◽  
Alkyl Ethers ◽  
New Compounds ◽  
Increased Activity

ABSTRACT Alkoxyalkyl esters of cidofovir (CDV) have substantially greater antiviral activity and selectivity than unmodified CDV against herpesviruses and orthopoxviruses in vitro. Enhancement of antiviral activity was also noted when cyclic CDV was esterified with alkoxyalkanols. In vitro antiviral activity of the most active analogs against human cytomegalovirus (HCMV) and orthopoxviruses was increased relative to CDV up to 1,000- or 200-fold, respectively. Alkyl chain length and linker structure are important potential modifiers of antiviral activity and selectivity. In this study, we synthesized a series of alkoxyalkyl esters of CDV or cyclic CDV with alkyl chains from 8 to 24 atoms and having linker moieties of glycerol, propanediol, and ethanediol. We also synthesized alkyl esters of CDV which lack the linker to determine if the alkoxyalkyl linker moiety is required for activity. The new compounds were evaluated in vitro against HCMV and murine CMV (MCMV). CDV or cyclic CDV analogs both with and without linker moieties were highly active against HCMV and MCMV, and their activities were strongly dependent on chain length. The most active compounds had 20 atoms esterified to the phosphonate of CDV. Both alkoxypropyl and alkyl esters of CDV provided enhanced antiviral activities against CMV in vitro. Thus, the oxypropyl linker moiety is not required for enhanced activity. CDV analogs having alkyl ethers linked to glycerol or ethanediol linker groups also demonstrated increased activity against CMV.

scholarly journals Phosphorimidate Derivatives of Acyclovir; Antiviral Activity Against Canine Parvovirus In Vitro

2019 ◽  
Author(s):  
Payal M Oak ◽  
Akash S. Mali ◽  
Atul R Chopade ◽  
Govind Shukla
Keyword(s):  
Antiviral Drug ◽  
Canine Parvovirus ◽  
Type Ii ◽  
Vitro Assay ◽  
Vaccination Programs ◽  
New Compounds ◽  
Derivatives Of ◽  
Micromolar Range ◽  
New Chemical Entity

AbstractCanine parvovirosis is a very transmissible, severe and often deadly infectious disease of dogs caused by Type II canine parvovirus (CPV-2). Recently shows that Parvoviruses are very resistant to Acyclovir which is often used for HSV Chemotherapy in humans and various virucidal purposes in animal diseases in veterinary clinics and animal housing facilities. If acquiescence with vaccination programs and with appropriate decontamination plans is guaranteed, there should be no continuous, nor common, CPV-2 outbreaks. However, a continuous spread of CPV-2 infections is observed, even in shelters where an appropriate vaccination program is applied, and this is reason to provide an antiviral drug therapy. The aim of the present study was to Development of antiviral drugs with determination the effect of concentration of new chemical entity and analyze Acyclovir analogous against CPV-2 strains. A sensitive in vitro assay capable of measuring the infectivity of CPV-2 was employed to determine the efficacy of three different concentrations of 9-(2-hydroxyethomethyl)guanine phosphoromonomorpholidate. We successfully show that new compounds inhibit CPV-2 replication with exhibiting 50% inhibitory concentrations (IC50s) in the low-micromolar range (50μM).

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