Fig. 7

Influence of targeted supplements on the decreased CSL performance after 23 months storage time, measured in a µTOM device. (A-B) Oxygen transfer rate of O. polymorpha RB11 pC10-FMD (P_FMD-GFP) on CSL medium enriched with different supplements. For clarity, the different supplements are grouped into two separate graphs (A and B). Shown in A and B are two reference CSL media containing CSL that was stored at different times: 16.5 months and 23 months, respectively. Except for the reference CSL medium containing CSL that was stored for 16.5 months, all other media contained 1.0 g/L ammonium sulfate. (A) Effect of different chemical solutions, based on Syn-6 synthetic MES medium, on CSL performance. (B) Influence of different compounds of the employed microelements solution on CSL performance. For clarity, the data for MnSO₄ and ZnSO₄ are not shown, as the course of the OTRs over time was nearly identical to the reference and the curve for FeSO₄, respectively. The storage time of the CSL batch employed in (A-B) was 23 months and the CSL batch was stored at 4 °C. Data points shown are mean values measured from 4–6 individual wells. Shadows, indicating the respective standard deviation, are barely visible, caused by the low standard deviations < 0.5. Experimental parameters: round deep-well 96-well microtiter plate (MTP), CSL 1 concentration \(\:{c}_{CSL,1}\:\)= 2.5 g dry substance/L, glucose concentration \(\:{c}_{glucose}\) = 5 g/L, ammonium sulfate concentration \(\:{c}_{(N{H}_{4}{)}_{2}S{O}_{4}}\) = 1.0 g/L, MES buffer concentration \(\:{c}_{MES}\) = 0.1 M, initial pH = 6.0, culture volume V_L = 200 µL/well, shaking frequency n = 350 rpm, shaking diameter d₀ = 50 mm, humidity = 80%, temperature T = 37 °C and initial OD₆₀₀ = 0.1