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>_m > 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