%0 Journal Article %1 ISI:000366406900002 %A Vallon, Tobias %A Simon, Oliver %A Rendgen-Heugle, Beate %A Frana, Sabine %A Mueckschel, Bjoern %A Broicher, Alexander %A Siemann-Herzberg, Martin %A Pfannenstiel, Jens %A Hauer, Bernhard %A Huber, Achim %A Breuer, Michael %A Takors, Ralf %C 111 RIVER ST, HOBOKEN 07030-5774, NJ USA %D 2015 %I WILEY-BLACKWELL %J ENGINEERING IN LIFE SCIENCES %K Butanol; Chemostat Omics; Stress culture; myown response} {Biofuels; %N 8 %P 760-771 %R 10.1002/elsc.201400051 %T Applying systems biology tools to study n-butanol degradation in Pseudomonas putida KT2440 %U https://doi.org/10.1002/elsc.201400051 %V 15 %X To smoothen the process of n-butanol formation in Pseudomonas putida KT2440, detailed knowledge of the impact of this organic solvent on cell physiology and regulation is of outmost importance. Here, we conducted a detailed systems biology study to elucidate cellular responses at the metabolic, proteomic, and transcriptional level. Pseudomonas putida KT2440 was cultivated in multiple chemostat fermentations using n-butanol either as sole carbon source or together with glucose. Pseudomonas putida KT2440 revealed maximum growth rates (mu) of 0.3 h(-1) with n-butanol as sole carbon source and of 0.4 h(-1) using equal C-molar amounts of glucose and n-butanol. While C-mole specific substrate consumption and biomass/substrate yields appeared equal at these growth conditions, the cellular physiology was found to be substantially different: adenylate energy charge levels of 0.85 were found when n-butanol served as sole carbon source (similar to glucose as sole carbon source), but were reduced to 0.4 when n-butanol was coconsumed at stable growth conditions. Furthermore, characteristic maintenance parameters changed with increasing n-butanol consumption. C-13 flux analysis revealed that central metabolism was split into a glucose-fueled Entner-Doudoroff/pentose-phosphate pathway and an n-butanol-fueled tricarboxylic acid cycle when both substrates were coconsumed. With the help of transcriptome and proteome analysis, the degradation pathway of n-butanol could be unraveled, thus representing an important basis for rendering P. putida KT2440 from an n-butanol consumer to a producer in future metabolic engineering studies.

@article{ISI:000366406900002, abstract = {{To smoothen the process of n-butanol formation in Pseudomonas putida KT2440, detailed knowledge of the impact of this organic solvent on cell physiology and regulation is of outmost importance. Here, we conducted a detailed systems biology study to elucidate cellular responses at the metabolic, proteomic, and transcriptional level. Pseudomonas putida KT2440 was cultivated in multiple chemostat fermentations using n-butanol either as sole carbon source or together with glucose. Pseudomonas putida KT2440 revealed maximum growth rates (mu) of 0.3 h(-1) with n-butanol as sole carbon source and of 0.4 h(-1) using equal C-molar amounts of glucose and n-butanol. While C-mole specific substrate consumption and biomass/substrate yields appeared equal at these growth conditions, the cellular physiology was found to be substantially different: adenylate energy charge levels of 0.85 were found when n-butanol served as sole carbon source (similar to glucose as sole carbon source), but were reduced to 0.4 when n-butanol was coconsumed at stable growth conditions. Furthermore, characteristic maintenance parameters changed with increasing n-butanol consumption. C-13 flux analysis revealed that central metabolism was split into a glucose-fueled Entner-Doudoroff/pentose-phosphate pathway and an n-butanol-fueled tricarboxylic acid cycle when both substrates were coconsumed. With the help of transcriptome and proteome analysis, the degradation pathway of n-butanol could be unraveled, thus representing an important basis for rendering P. putida KT2440 from an n-butanol consumer to a producer in future metabolic engineering studies.}}, added-at = {2018-01-25T13:38:08.000+0100}, address = {{111 RIVER ST, HOBOKEN 07030-5774, NJ USA}}, affiliation = {{Takors, R (Reprint Author), Univ Stuttgart, Inst Biochem Engn, Allmandring 31, D-70569 Stuttgart, Germany. Vallon, Tobias; Rendgen-Heugle, Beate; Frana, Sabine; Broicher, Alexander; Siemann-Herzberg, Martin; Takors, Ralf, Univ Stuttgart, Inst Biochem Engn, D-70569 Stuttgart, Germany. Simon, Oliver; Pfannenstiel, Jens; Huber, Achim, Univ Hohenheim, Life Sci Ctr, Prote Core Facil, Stuttgart, Germany. Mueckschel, Bjoern; Hauer, Bernhard, Univ Stuttgart, Inst Tech Biochem, D-70569 Stuttgart, Germany. Breuer, Michael, BASF SE, Fine Chem \& Biocatalysis Res, Ludwigshafen, Germany.}}, author = {Vallon, Tobias and Simon, Oliver and Rendgen-Heugle, Beate and Frana, Sabine and Mueckschel, Bjoern and Broicher, Alexander and Siemann-Herzberg, Martin and Pfannenstiel, Jens and Hauer, Bernhard and Huber, Achim and Breuer, Michael and Takors, Ralf}, author-email = {{takors@ibvt.uni-stuttgart.de}}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2652fb4ff4b5092a491db6399dff0fe9d/siemannherzberg}, da = {{2018-01-25}}, doc-delivery-number = {{CY4VM}}, doi = {{10.1002/elsc.201400051}}, eissn = {{1618-2863}}, funding-acknowledgement = {{German Federal Ministry of Education and Research (BMBF) {[}FKZ: 0315406]; BASF SE}}, funding-text = {{This study was supported by the German Federal Ministry of Education and Research (BMBF; FKZ: 0315406) and BASF SE.}}, interhash = {4b90fac7e2fb842634a098f46bc45a2b}, intrahash = {652fb4ff4b5092a491db6399dff0fe9d}, issn = {{1618-0240}}, journal = {{ENGINEERING IN LIFE SCIENCES}}, journal-iso = {{Eng. Life Sci.}}, keywords = {Butanol; Chemostat Omics; Stress culture; myown response} {Biofuels;}, keywords-plus = {{AROMATIC CATABOLIC PATHWAYS; COMPLETE GENOME SEQUENCE; IN-VIVO ANALYSIS; SP STRAIN VLB120; SOLVENT-TOLERANT; TOLUENE METABOLISM; PROTEOMIC ANALYSIS; CARBON METABOLISM; STATISTICAL-MODEL; MASS-SPECTROMETRY}}, language = {{English}}, month = {{NOV}}, number = {{8}}, number-of-cited-references = {{55}}, pages = {{760-771}}, publisher = {{WILEY-BLACKWELL}}, research-areas = {{Biotechnology \& Applied Microbiology}}, times-cited = {{6}}, timestamp = {2018-01-25T12:38:18.000+0100}, title = {{Applying systems biology tools to study n-butanol degradation in Pseudomonas putida KT2440}}, type = {{Article}}, unique-id = {{ISI:000366406900002}}, url = {https://doi.org/10.1002/elsc.201400051}, usage-count-last-180-days = {{2}}, usage-count-since-2013 = {{7}}, volume = {{15}}, web-of-science-categories = {{Biotechnology \& Applied Microbiology}}, year = {{2015}} }