Extended Data Fig. 1: Target screening for co-culture potential using 52 pairs of adenine-exchanged metabolite cross-feeding co-cultures.
From: A molecular toolkit of cross-feeding strains for engineering synthetic yeast communities
a. 52 pairs of adenine-exchanged metabolite (ade-em) two-member cross-feeding co-cultures were created for each target, including ade-em I, overexpression of em and ade in each member; ade-em II, overexpression of ade only in blue; ade-em III, overexpression of em only in red; ade-em IV, no overexpression in each member. Em includes arg, cys, his, leu, lys, met, phe, tyr, ser, thr, trp, ura, val, ile. BY4741-pHLUM was used as the positive control (+ve). These co-cultures were tested with the initial ratio of 1:1 and initial cell density of each member was OD700nm 0.078 by a Tecan Spark plate reader. b. OD700nm values of negative controls for monocultures at 72 h. The strain adeΔem (red) was duplicated in the diagram to correspond with the numbers of co-culture combinations. c. Estimated maximum OD700nm values of red population (ROD), blue population (BOD) and total OD values (ROD + BOD) in ade-em co-cultures within 72 h. The standard curves of estimating the fluorescent intensities of BFP and RFP to OD values were shown in Supplementary Table 3. d. The maximal measured total OD700nm values of 52 pairs of adenine-exchanged metabolite co-cultures at 72 h are close to the sum of estimated values of BOD and ROD. N = three biologically independent samples, and data are presented as mean values +/− SD. Two-way ANOVA, followed by Bonferroni’s multiple comparisons test with 95% confidence intervals were performed using Prism 9.5.0 (GraphPad) software, and p values were noted.
