%0 Journal Article %1 ISI:000327843200005 %A Blombach, Bastian %A Buchholz, Jens %A Busche, Tobias %A Kalinowski, Joern %A Takors, Ralf %C PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS %D 2013 %I ELSEVIER SCIENCE BV %J JOURNAL OF BIOTECHNOLOGY %K myown %N 4 %P 331-340 %R 10.1016/j.jbiotec.2013.10.005 %T Impact of different CO2/HCO3- levels on metabolism and regulation in Corynebacterium glutamicum %U https://doi.org/10.1016/j.jbiotec.2013.10.005 %V 168 %X 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.

@article{ISI:000327843200005, abstract = {{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.}}, added-at = {2018-06-08T13:06:36.000+0200}, address = {{PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}}, affiliation = {{Takors, R (Reprint Author), Univ Stuttgart, Inst Biochem Engn, Allmandring 31, D-70569 Stuttgart, Germany. Blombach, Bastian; Buchholz, Jens; Takors, Ralf, Univ Stuttgart, Inst Biochem Engn, D-70569 Stuttgart, Germany. Busche, Tobias; Kalinowski, Joern, Ctr Biotechnol CeBiTec Microbial Genom \& Biotechn, D-33615 Bielefeld, Germany.}}, author = {Blombach, Bastian and Buchholz, Jens and Busche, Tobias and Kalinowski, Joern and Takors, Ralf}, author-email = {{blombach@ibvt.uni-stuttgart.de takors@ibvt.uni-stuttgart.de}}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/28b4cbd032db5253898ef69405b0a4533/ralftakors}, da = {{2018-01-26}}, doc-delivery-number = {{263YF}}, doi = {{10.1016/j.jbiotec.2013.10.005}}, eissn = {{1873-4863}}, funding-acknowledgement = {{Deutsche Forschungsgemeinschaft (DFG) {[}TA 241/5-1]}}, funding-text = {{We thank Mira Lenfers-Lucker for the assistance with the HPLC analyses, Michaela Graf, Michael Ghosh, and Andreas Ulmer for the support in the bioreactor studies and the strain construction, Michael Kraml and Salaheddine Laghrami for the technical support, as well as Andreas Freund for the technical advice in carrying out the fermentation experiments. We also thank Eva Schulte-Berndt for the technical assistance in the microarray analyses. The authors further gratefully acknowledge the funding of this work by the Deutsche Forschungsgemeinschaft (DFG), grant TA 241/5-1.}}, interhash = {75535e85947aaa1c453c03c26dfd2dfc}, intrahash = {8b4cbd032db5253898ef69405b0a4533}, issn = {{0168-1656}}, journal = {{JOURNAL OF BIOTECHNOLOGY}}, journal-iso = {{J. Biotechnol.}}, keywords = {myown}, keywords-plus = {{DISSOLVED CARBON-DIOXIDE; PYRUVATE-QUINONE OXIDOREDUCTASE; RECOMBINANT ESCHERICHIA-COLI; AMINO-ACIDS; BIOCHEMICAL-CHARACTERIZATION; FERMENTATION PROCESSES; DEHYDROGENASE COMPLEX; NUTRIENT CONSUMPTION; PHYLOGENETIC ASPECTS; MOLECULAR ANALYSIS}}, language = {{English}}, month = {{DEC}}, number = {{4}}, number-of-cited-references = {{68}}, pages = {{331-340}}, publisher = {{ELSEVIER SCIENCE BV}}, research-areas = {{Biotechnology \& Applied Microbiology}}, times-cited = {{12}}, timestamp = {2018-06-08T13:06:36.000+0200}, title = {{Impact of different CO2/HCO3- levels on metabolism and regulation in Corynebacterium glutamicum}}, type = {{Article}}, unique-id = {{ISI:000327843200005}}, url = {https://doi.org/10.1016/j.jbiotec.2013.10.005}, usage-count-last-180-days = {{2}}, usage-count-since-2013 = {{14}}, volume = {{168}}, web-of-science-categories = {{Biotechnology \& Applied Microbiology}}, year = {{2013}} }