ABSTRACT
The glycosylation of natural product scaffolds with highly modified deoxysugars is often essential for their biological activity, being responsible for specific contacts to molecular targets and significantly affecting their pharmacokinetic properties. In order to provide tools for the targeted alteration of natural product glycosylation patterns, significant strides have been made to understand the biosynthesis of activated deoxysugars and their transfer. We report here efforts towards the production of plasmid-borne biosynthetic gene cassettes capable of producing TDP-activated forms of D-mycaminose, D-angolosamine and D-desosamine. We additionally describe the transfer of these deoxysugars to macrolide aglycones using the glycosyl transferases EryCIII, TylMII and AngMII, which display usefully broad substrate tolerance.
Subject(s)
Glucosamine/analogs & derivatives , Macrolides/chemistry , Macrolides/metabolism , Cloning, Molecular , Genetic Engineering , Glucosamine/chemistry , Glucosamine/metabolism , Molecular Structure , Multigene Family/genetics , Sequence Analysis , Streptomyces/chemistry , Streptomyces/genetics , Streptomyces/metabolismABSTRACT
An acetophenone containing PNA-based reagent was designed for the direct and site-specific synthesis of a cis-syn thymidine dimer lesion in genomic DNA.
Subject(s)
Acetophenones/chemistry , DNA/chemical synthesis , Peptide Nucleic Acids/chemistry , Pyrimidine Dimers/chemical synthesis , Acetophenones/chemical synthesis , Bacteriophages/genetics , Base Sequence , DNA/chemistry , DNA/genetics , Deoxyribonucleases, Type II Site-Specific/chemistry , Horseradish Peroxidase/chemistry , Molecular Sequence Data , Streptavidin/chemistry , Substrate SpecificityABSTRACT
PNA:DNA strands were prepared containing a flavin electron donor and a thymine dimer acceptor, which gives a strand break upon single electron reduction. With these constructs, it was confirmed that an excess electron transfer through the base stack can be efficient in an interstrand fashion. The effect of an increased distance, a changed sequence, and stacking was explored.
