Supplementing mixture X (TmXyn10A xylanase and BpXyn43 β-xylosidase) to mixture C led to a significant increase (by 25to71%1%, t-test: P < 0.01) in xylose release from the pith of the H58 clone with lower lignin content, whereas the increase was less significant (t-test: 0.01 < P < 0.03 up to 48 h) in the interface fraction of H58.
Richard Wurtman invented the supplement mixture, known as Souvenaid, which appears to stimulate growth of new synapses.
The results suggested that a dietary supplement mixture of 0.7% arginine and 1% glutamine improved the growth performance and the intestinal mucosa development of weaned piglets and that the effect was greater than with the single supplementation of either arginine or glutamine.
Agar plates of synthetic medium (6.7 g/l yeast nitrogen base w/o aminoacids, 0.75 g/l complete supplement mixture w/o uracil, 2% glucose, 2% agar, pH 6) or YPD (2% glucose, 2% peptone, 1% yeast extract, 2% agar) supplemented with 200 mg/l of G418, were used for selection of the strains.
Selection and cultivation of yeast transformants were conducted on Synthetic Dextrose (SD) medium lacking uracil (SD-URA plates: 6.9 g/L yeast nitrogen base (YNB) without amino acids (Formedium), 0.77 g/L complete supplement mixture without uracil (Formedium), 20 g/L glucose and 20 g/L agar).
Counterselection of the URA3 marker was performed by cultivation on 5-fluoroorotic acid containing plates [5-FOA plates: 6.9 g/L YNB without amino acids (Formedium), 0.77 g/L complete supplement mixture (Formedium), 20 g/L glucose and 0.8 g/L 5-fluoroorotic acid (Sigma)].
Plasmid containing yeast strains were selected on synthetic dextrose (SD) medium, prepared with 6.7 g L-1 yeast nitrogen base without amino acids (YNB-AA) (Formedium, Hunstanton, UK) and 20 g L-1 glucose with complete supplement mixture (CSM) lacking uracil and/or histidine (Formedium) where appropriate.
Complete synthetic medium contained 0.67% yeast nitrogen base without amino acids (YNB), 1× complete supplement mixture (CSM) (Bio 101), and 2% glucose.
Saccharomyces cerevisiae cells BY4741 ssk1Δ (BY4741; Mat a; his3Δ1; leu2Δ0; met15Δ0; ura3Δ0; ssk1::kanMX4, from the Saccharomyces Genome Deletion Project) were grown in synthetic complete medium (1x Difco™ YNB base, 1x Formedium™ Complete Supplement Mixture, 0.5% ammonium sulfate, 2% glucose) on a rotary shaker at 225 rpm at 30°C until reaching an optical density of 1.0 measured at 600 nm.
This strain was transformed by various pRS416-derived plasmids using a simplified lithium acetate method (Gietz et al, 1992) and selected on complete supplement mixture lacking uracil (CSM-ura; a synthetic medium supplemented with CSM) containing 2% glucose.
