Methyltransferase Contingencies in the Pathway of Everninomicin D Antibiotics and Analogues

Everninomicins are orthoester oligosaccharide antibiotics with potent activity against multidrug-resistant bacterial pathogens. Everninomicins act by disrupting ribosomal assembly in a distinct region in comparison to clinically prescribed drugs. We employed microporous intergeneric conjugation with Escherichia coli to manipulate Micromonospora for targeted gene-replacement studies of multiple putative methyltransferases across the octasaccharide scaffold of everninomicin effecting the A₁, C, F, and H rings. Analyses of gene-replacement and genetic complementation mutants established the mutability of the everninomicin scaffold through the generation of 12 previously unreported analogues and, together with previous results, permitted assignment of the ten methyltransferases required for everninomicin biosynthesis. The in vitro activity of A₁- and H-ring-modifying methyltransferases demonstrated the ability to catalyze late-stage modification of the scaffold on an A₁-ring phenol and H-ring C-4’ hydroxy moiety. Together these results establish the potential of the everninomicin scaffold for modification through mutagenesis and in vitro modification of advanced biosynthetic intermediates.     

Controllability Analysis of Process Alternatives for Biobutanol Purification

Biobutanol has characteristics similar to petroleum fuel and is considered as a superior biofuel compared to ethanol. The development of technologies for biobutanol production by fermentation has resulted in higher final biobutanol concentrations together with less energy‐intensive separation and purification techniques. These new technological developments have the potential to provide a production process for biobutanol that is economically viable in comparison to the petrochemical pathway for its production. The control properties of four different possible process designs for biobutanol purification are analyzed. The results, using the singular value decomposition technique, indicated that the scheme where only biobutanol flow is purified, and both ethanol and acetone leaving the purification process mixed with water and biobutanol traces, showed the best control properties.