Journal: Nature Chemical Biology
Article Title: Spontaneously established syntrophic yeast communities improve bioproduction
doi: 10.1038/s41589-023-01341-2
Figure Lengend Snippet: ( a ) BY4741-derived deletion strains were complemented with pHLUM to partially restore wild-type alleles. Since BY4741-derived strains containing pHLUM do not require histidine, leucine, methionine, or uracil to grow in minimal media, any observed growth deficiencies could be linked to other uncomplemented deletion mutations. ( b ) Starting from 5,185 mutants, the library was initially refined to 157, based on previous experimental observations from Mülleder et al. and then to 92 strains, based on an experiment to measure the growth characteristics of the 157 strains in synthetic minimal media. ( c ) BY4741-derived mutants transformed with pHLUM were cultivated in synthetic minimal media for 18 h. Strains were compared with two known prototrophs: BY4741-derived strains with met17∆ or his3∆ deletion mutations (that is, BY4741 mutants with all auxotrophic markers restored by pHLUM and thus no fitness disadvantage; green bars). The red dotted red line represents 20% of average prototrophic growth, which was used as the threshold to choose between mutants that could not grow in synthetic minimal media (purple) and were therefore suitable for the pairwise auxotroph complementation screen and those that were unsuitable (light brown) (see Fig. and Extended Data Figs. – ). Error bars (panel c) denote standard error around the mean.
Article Snippet: OD of all five output plates was measured again at 48 h. Strains were grown in YPD (2% (wt/vol) glucose (Sigma-Aldrich, G8270), 20 g l −1 peptone (Bacto, 211677) and 10 g l −1 yeast extract (Bacto, 212750)); SM media (2% (wt/vol) glucose and 6.7 g l −1 yeast nitrogen base without amino acids (Sigma-Aldrich, Y1251)); SC-His (2% (wt/vol) glucose, 6.8 g l −1 yeast nitrogen base, 0.56 g l −1 CSM-His-Leu-Met-Trp-Ura (powder; MP Biomedicals), 60 mg l −1 leucine, 20 mg l −1 methionine, 40 mg l −1 tryptophan and 20 mg l −1 uracil) or SC-Ura (2% (wt/vol) glucose, 6.8 g l −1 yeast nitrogen base, 0.56 g l −1 CSM-His-Leu-Met-Trp-Ura (powder; MP Biomedicals, 114550422), 60 mg l −1 leucine (Thermo Fisher Scientific, AC125125000), 20 mg l −1 methionine (Thermo Fisher Scientific, A1031836), 40 mg l −1 tryptophan (Thermo Fisher Scientific, 140591000) and 20 mg l −1 histidine (Thermo Fisher Scientific, 166155000)).
Techniques: Derivative Assay, Transformation Assay
