For evaluation of the fermentation performance in medium containing D-xylose and glucose, the batch fermentations were performed in complex medium in 300 mL shake flasks with a working volume of 200 mL at 35°C.
The feeding medium [ 17] contained D-glucose at an initial concentration of 3 g/L, a value low enough to make the culture carbon-limited.
After 120 h, the medium was replaced by fresh medium containing D-luciferin (150 μg/ml).
The respective DS68625 strains were pre-grown on 2% maltose and transferred to minimal medium containing 2% D-glucose or D-xylose.
To validate observations from the bioreactor cultures, the strains were grown in 1 L Erlenmeyer flasks in 400 mL of YP medium containing 10 g/L D-glucose and 20 g/L D-xylose at 30°C, 250 rpm.
The rat insulinoma INS-1 cells were cultured in RPMI-1640 medium containing 1 mM d-glucose and supplemented with 10%% fetal calf serum, 100 U/mL penicillin, 100 mg/mL streptomycin, 10 M HEPES, 2 M l-glutamine, 1 M sodium pyruvate, and 50 μM β-mercaptoethanol [20].
On the other hand, in HUVEC incubated in a medium containing 5.5 mmol/L D-glucose, the pro-inflammatory cytokine IL-1β enhanced the cell surface expression of both ICAM-1 and VCAM-1 in a concentration-dependent manner, with a sub-maximal effect observed at 5 ng/mL (Figures 1A-1B).
Cells were thawed and propagated in DMEM (Dulbecco's modified Eagle's medium) containing 4.5 g/l D-glucose and 10% fetal bovine serum, and three passages later frozen in aliquots of 1 ml.
HEK293 and U2OS cells were cultured in Dulbecco's-modified Eagle's medium, containing 4.5 g/l D-glucose and GlutaMAX (Life Technologies, Carlsbad, CA, USA), and supplemented with 10% fetal bovine serum.
(A) T. reesei QM6aΔ tmus53 (blue) and the xpp1 deletion strain (green) were grown in MA medium containing 1% (w/v) D-glucose (squares, solid lines) or lactose (triangles, dashed lines), or CMC (circles, dotted lines) for 18, 24, 30, 38, and 50 hours.
