Impact of different CO2/HCO3- levels on metabolism and regulation in Corynebacterium glutamicum

We investigated the growth kinetics and transcriptional responses of Corynebacterium glutamicum in environments with low (pCO(2) < 40 mbar) and high (pCO(2) >= 300 mbar) CO2/HCO3 levels compared to standard conditions. When cultivated at high CO2/HCO3--levels, C. glutamicum showed increased (63%) biomass to substrate yields during the initial growth phase. Other kinetic parameters such as growth rate (mu), specific glucose consumption rate (q(s)), and selected enzymatic activities of anaplerotic reactions, the pentose phosphate pathway and the tricarboxylic acid cycle were similar to standard conditions. However, microarray hybridization disclosed a complex transcriptional response involving 117 differentially expressed genes. Among those, 60 genes were assigned to the complete DtxR/RipA regulon controlling iron homeostasis in C. glutamicum. Impaired growth of a Delta dtxR mutant at high CO2/HCO3- levels validated the relevance of this master regulator to cope with excessive CO2/HCO3- availability. At low CO2/HCO3- levels, C. glutamicum grew in a bi-level manner with three distinct growth phases. Differential analyses revealed approximately doubled activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase accompanied by the formation of L-alanine and L-valine during the lowest mu occurring in mid-phase of the cultivation. DNA microarray analysis revealed more than 100 differentially expressed genes in growth phase II compared to phase I including almost all thiamin pyrophosphate (TPP) biosynthesis genes, which were significantly up regulated. Concluding, we hypothesize that C. glutamicum counteracts the lack of CO2/HCO3- by triggering TPP biosynthesis for increasing the activities of TPP-dependent enzymes involved in CO2 formation. (C) 2013 Elsevier B.V. All rights reserved.