Adipic acid is one of the most important
small molecules in the
modern chemical industry. However, the damaging environmental impact
of the current industrial synthesis of adipic acid has necessitated
the development of greener, biobased approaches to its manufacture.
Herein we report the first one-pot synthesis of adipic acid from guaiacol,
a lignin-derived feedstock, using genetically engineered whole-cells
of Escherichia coli. The reaction is mild, efficient,
requires no additional additives or reagents, and produces no byproducts.
This study demonstrates how modern synthetic biology can be used to
valorize abundant feedstocks into industrially relevant small molecules
in living cells.
Surface
plasmon resonance (SPR) is a widely used method to study
ligand–protein interactions. The throughput and sensitivity
of SPR has made it an important technology for measuring low-affinity,
ultralow weight fragments (<200 Da) in the early stages of drug
discovery. However, the biochemistry of membrane proteins, such as
G-protein-coupled receptors (GPCRs), makes their SPR fragment screening
particularly challenging, especially for native/wild-type, nonthermostabilized
mutant receptors. In this study, we demonstrate the use of SPR-based
biosensors to study the entire human family of adenosine receptors
and present biologically active novel binders with a range of selectivity
to human adenosine 2a receptor (hA2AR) from an ultralow
weight fragment library and the public GlaxoSmithKline (GSK) kinase
library. Thus, we demonstrate the ability of SPR to screen ultra-low-affinity
fragments and identify biologically meaningful chemical equity and
that SPR campaigns are highly effective “chemical filters”
for screening small building block fragments that can be used to enable
drug discovery programs.
The fundamental biology
and application of bacterial exopolysaccharides
is gaining increasing attention. However, current synthetic biology
efforts to produce the major component of Escherichia sp. slime, colanic acid, and functional derivatives thereof have been
limited. Herein, we report the overproduction of colanic acid (up
to 1.32 g/L) from d-glucose in an engineered strain of Escherichia coli JM109. Furthermore, we report that chemically
synthesized l-fucose analogues containing an azide motif
can be metabolically incorporated into the slime layer via a heterologous
fucose salvage pathway from Bacteroides sp. and used
in a click reaction to attach an organic cargo to the cell surface.
This molecular-engineered biopolymer has potential as a new tool for
use in chemical, biological, and materials research.
The fundamental biology and application of bacterial exopolysaccharides is gaining increasing attention. However, current synthetic biology efforts to produce the major component of Escherichia sp. slime, colanic acid, and functional derivatives thereof have been limited. Herein, we report the overproduction of colanic acid (up to 1.32 g/L) from D-glucose in an engineered strain of E. coli JM109. Furthermore, we report that chemically-synthesized L-fucose analogues containing an azide motif can be metabolically incorporated into the slime layer via a heterologous fucose salvage pathway from Bacteroides sp. and used in a click reaction to attach an organic cargo to the cell surface. This molecular engineered bio-polymer possesses enormous potential as a new tool for use in chemical, biological and materials research.
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