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Supplementary information
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Quantitative physiology of Lactococcus lactis at extreme low-growth rates
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Onur Ercan1,2,3,4, Eddy J. Smid2,6, Michiel Kleerebezem2,4,5,*
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Running title: L. lactis zero-growth physiology
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Keywords: Lactococcus lactis, zero-growth, quantitative physiology, maintenance energy, modeling
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Kluyver Centre for Genomics of Industrial Fermentation, P.O. Box 5057, 2600 GA Delft, The
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Netherlands,
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Top Institute Food and Nutrition, Nieuwe Kanaal 9A, 6709 PA Wageningen, The Netherlands,
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Laboratory of Microbiology, Wageningen University, P.O. Box 8033, 6700 EJ Wageningen, The
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Netherlands,
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NIZO food research, P.O. Box 20, 6710 BA Ede, The Netherlands,
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Host Microbe Interactomics, Wageningen University, P.O. Box 338, 6700 AH Wageningen, The
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Netherlands,
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The Netherlands.
Laboratory of Food Microbiology, Wageningen University, P.O. Box 8129, 6700 EV Wageningen,
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Correspondence: Michiel Kleerebezem, Host Microbe Interactomics Group, Wageningen University,
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De Elst 1, 6708 WD Wageningen, The Netherlands. Phone: +31-317-486125; Fax: +31-317-483962;
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E-mail: michiel.kleerebezem@wur.nl
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Day 35
Day 42
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Fig. S1: Flow cytometric analysis of L. lactis KF147 in samples taken on days 35 and 42 under
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retentostat cultivation. Bacterial cells were stained with a mixture of SYTO9 and PI, and then analyzed
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at red fluorescence (PE-Texas Red-A) versus green fluorescence (FITC-A) on a flow cytometer.
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B
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Fig. S2: Kinetics of specific ATP production and carbon consumption rates in retentostat cultures of L.
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lactis KF147. A steady-state anaerobic chemostat culture was switched to retentostat mode at time
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zero. (A) Specific ATP production rate (qATP) and (B) specific carbon consumption rate (qcarbon). The
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dashed lines indicate the average of energy (mATP)- and substrate-dependent maintenance coefficient
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(ms), respectively, calculated from retentostat 1 (diamonds) and retentostat 2 (squares) cultivations.
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Fig. S3: Plot of the specific substrate consumption rate qs against μ in anaerobic, glucose-limited
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chemostat cultures of L. lactis KF147. Each data point represents the average of data from three
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different chemostat cultures.
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Day 0
Day 21
Day 2
Day 28
Day 7
Day 35
Day 14
Day 42
Fig. S4: Bright field images of L. lactis KF 147 cells from retentostat 1 in the beginning of the
retentostat, days 2, 7, 14, 21, 28, 35, and 42. Scale bar 1 cm.
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