Briefly, isolated bright cells located over the culture monolayer of culture flasks were obtained after rinsing and manual shaking, followed by centrifugation of the supernatant at 350 g for 5 min. After resuspension, microglial cells were plated and maintained in glial medium as described previously.
All culture plates and flasks were obtained from TPP (Zolstrasse, Trasadingen, Switzerland).
For 3D6scFv-Fc, the maximal product yield in shake flask screening cultures was obtained from a strain which was selected on 500 μg mL 1 Zeocin (Supporting information, Fig. S1A).
After two rounds of genome shuffling, a high-yield recombinant YSF2-19 sthatn that exhibited 3.2- and 3.3-fold increases in glutathione production in shake flask and fermenter respectively was obtained.
A glycerol concentration of 6.8 g l−1 was obtained in flask culture.
Cells were grown in T25 flasks until 60% of confluency was obtained.
From 11 different compositions of dual co-cultures grown on Avicel in flasks, the highest ethanol production was obtained with a co-culture comprising Caldicellulosiruptor DIB 087C and Thermoanaerobacter DIB 097X: 13.8 mM ethanol and 34.8 mol% of ethanol fraction within the total organic products.
Interestingly, only 0.17 mol% 4HB in the copolymer was obtained during shake flask studies.
Under these best suited conditions, a concentration of 11g/L of arabitol and 19 g/L of ethanol was obtained in shake flask fermentations.
For each substrate, C count as the background value was obtained from a flask without cells.
POx was recombinantly produced in E. coli as reported in ref. [ 14], while CDH was obtained from shaken flask cultures of P. pastoris basically as described by Sygmund [ 15], however, omitting the trace element solution from the medium.
