Fig. 4: Results of time-dependent batch experiments.
From: Timed batch inputs unlock substantially higher yields for enzymatic cascades
a, Three time series showing each addition of input species within the ERN: HK, G6PDH, 6PGDH, PRI, PRPPS, ATP, NADP and GLU. The y axis shows the volume of the additions to the reactor in µl and the x axis the time of additions. The algorithm allows an input to be given every 5 min, and each experiment (1–3) samples from different stock solutions. b, The time series plots show the prediction of the model for the time-dependent batch experiment (orange) and its corresponding control (green), (n = 10, 95% confidence intervals). c, A bar plot of the concentrations in μM of one of the final products AMP as measured on the HPLC. The time-dependent batch experiments (orange) have substantially higher final AMP concentrations compared with the control (green) as predicted in b and final concentration with a 5.6-fold increase compared with the control for experiment 3 (number above orange bar). Additional control experiments show that the spontaneous ATP hydrolysis does not affect outcomes (Supplementary Fig. 13). d, The yields indicate that a notably larger percentage of the initial substrate, glucose, was converted to AMP in all experiments compared with the control (yield = 100 n(AMP)final/n(glucose)added). Notably, in experiment 2, nearly half of the input substrate was transformed into the product.
