Abstract
The present work presents a novel approach for the dynamic quantification of respiration rates on a small scale by using lysine-producing Corynebacterium glutamicum ATCC 21253. Cells sampeld from batch cultures at different times were incubated ina 12-ml scale bioreactor equipped with a membrane mass spectrometer. Under dynamic conditions, gas exchange across the gas-liquid phase, specific respiration rates, and RQ values were precisely measured. For this purpose, suitable mass balances were formulated. The transport coefficients for $O_2$ and $CO_2$, crucial for calculating the respiration activity, were determined as $k_La_{O2}=9.18h^{-1}$ and $k_La_{CO2}=5.10h^{-1}$ at 400 rpm. The application of the proposed method to batch cultures of C. glutamicum ATCC 21253 revealed the maximum specific respiration rates of $q_{O2}=8.4\;mmol\;g^{-1}h^{-1}\;and\;q_{CO2}=8.7\;mmol\;g^{-1}h^{-1}$ in the middle of the exponential growth phase after 5 h of cultivation. When the cells changed from growth to lysine production due to the depletion of the essential amino acids theonine, methionine, and leucine, $q_{O2}\;and\;q_{CO2}$ decreased significantly and RQ increased. The respiration data exhibited an excellent agreement with previous cultivations of the strain [13]. This confirms the potential of the developed approach to realistically reflect the metabolic activities of cells at their point of sampling. The short-term influence of added threonine, methionine, and leucine was highest during the shift from growth to lysine production, where $q_{O2}\;and\;q_{CO2}$ increased 50% within one minute after the pulse addition of these compounds. Non-growing, yet lysine-producing cells taken from the end of the batch cultivation revealed no metabolic stimulation with the addition of threonine, methionine, and leucine.