5 research outputs found
MOESM2 of High yield 1,3-propanediol production by rational engineering of the 3-hydroxypropionaldehyde bottleneck in Citrobacter werkmanii
Additional file 2: Sequence data confirming different knock-out strains. Green = sequence of P1 primer; blue = sequence of P2 primer; red = FRT scar; purple = chloramphenicol resistance gene
Biphasic Catalysis with Disaccharide Phosphorylases: Chemoenzymatic Synthesis of α‑d‑Glucosides Using Sucrose Phosphorylase
Thanks
to its broad acceptor specificity, sucrose phosphorylase (SP) has
been exploited for the transfer of glucose to a wide variety of acceptor
molecules. Unfortunately, the low affinity (<i>K</i><sub>m</sub> > 1 M) of SP towards these acceptors typically urges the
addition of cosolvents, which often either fail to dissolve sufficient
substrate or progressively give rise to enzyme inhibition and denaturation.
In this work, a buffer/ethyl acetate ratio of 5:3 was identified to
be the optimal solvent system, allowing the use of SP in biphasic
systems. Careful optimization of the reaction conditions enabled the
synthesis of a range of α-d-glucosides, such as cinnamyl
α-d-glucopyranoside, geranyl α-d-glucopyranoside,
2-<i>O</i>-α-d-glucopyranosyl pyrogallol,
and series of alkyl gallyl 4-<i>O</i>-α-d-glucopyranosides. The usefulness of biphasic catalysis was further
illustrated by comparing the glucosylation of pyrogallol in a cosolvent
and biphasic reaction system. The acceptor yield for the former reached
only 17.4%, whereas roughly 60% of the initial pyrogallol was converted
when using biphasic catalysis
MOESM1 of Petroselinic acid purification and its use for the fermentation of new sophorolipids
Additional file 1: Fig. S1. 1H-NMR spectrum for compound 1. Fig. S2. 13C-NMR spectrum for compound 1. Fig. S3. 1H-NMR spectrum for compound 5. Fig. S4. 13C-NMR spectrum for compound 5. Fig. S5. 1H-NMR spectrum for compound 7. Fig. S6. 13C-NMR spectrum for compound 7. Fig. S7. 1H-NMR spectrum for compound 8. Fig. S8. 13C-NMR spectrum for compound 8. Fig. S9. 1H-NMR spectrum for compound 9. Fig. S10. 13C-NMR spectrum for compound 9. Fig. S11. 1H-NMR spectrum for compound 4. Fig. S12. 13C-NMR spectrum for compound 4
SILAC-Based Proteome Analysis of <i>Starmerella bombicola</i> Sophorolipid Production
<i>Starmerella (Candida) bombicola</i> is the biosurfactant-producing
species that caught the greatest deal of attention in the academic
and industrial world due to its ability of producing large amounts
of sophorolipids. Despite its high economic potential, the biochemistry
behind the sophorolipid biosynthesis is still poorly understood. Here
we present the first proteomic characterization of <i>S. bombicola</i> for which we created a <i>lys1</i>Δ mutant to allow
the use of SILAC for quantitative analysis. To characterize the processes
behind the production of these biosurfactants, we compared the proteome
of sophorolipid producing (early stationary phase) and nonproducing
cells (exponential phase). We report the simultaneous production of
all known enzymes involved in sophorolipid biosynthesis including
a predicted sophorolipid transporter. In addition, we identified the
heme binding protein Dap1 as a possible regulator for Cyp52M1. Our
results further indicate that ammonium and phosphate limitation are
not the sole limiting factors inducing sophorolipid biosynthesis
Topological Connection between Vesicles and Nanotubes in Single-Molecule Lipid Membranes Driven by Head–Tail Interactions
Lipid
nanotube–vesicle networks are important
channels for
intercellular communication and transport of matter. Experimentally
observed in neighboring mammalian cells but also reproduced in model
membrane systems, a broad consensus exists on their formation and
stability. Lipid membranes must be composed of at least two molecular
components, each stabilizing low (generally a phospholipid) and high
curvatures. Strong anisotropy or enhanced conical shape of the second
amphiphile is crucial for the formation of nanotunnels. Anisotropic
driving forces generally favor nanotube protrusions from vesicles.
In this work, we report the unique case of topologically connected
nanotubes–vesicles obtained in the absence of directional forces,
in single-molecule membranes, composed of an anisotropic bolaform
glucolipid, above its melting temperature, Tm. Cryo-TEM and fluorescence confocal microscopy show the interconnection
between vesicles and nanotubes in a single-phase region, between 60
and 90 °C under diluted conditions. Solid-state NMR demonstrates
that the glucolipid can assume two distinct configurations, head–head
and head–tail. These arrangements, seemingly of comparable
energy above the Tm, could explain the
existence and stability of the topologically connected vesicles and
nanotubes, which are generally not observed for classical single-molecule
phospholipid-based membranes above their Tm