A transcriptional study of acidogenic chemostat cells of Clostridium acetobutylicum following a single n-butanol pulse
ABSTRACT: In this study the transcriptional behavior of the natural solvent producing bacterium Clostridium acetobutylicum was investigated following n-butanol stress using DNA microarray analysis. Therefore, a phosphate-limited chemostat culture was established and n-butanol stress (0.9%) was added to acidogenic cells at pH 5.7.
Project description:Clostridium acetobutylicum is characterized by its acetone-butanol (AB) fermentation which <br>can be reproducibly established under continuous grow conditions in a chemostat. <br>At pH 5.7 cells show typical acidogenic metabolism and mainly produce the acids <br>acetate and butyrate. After lowering and further control the external pH at 4.5 <br>the exponentially growing cells switch towards stable solvent production with the <br>dominating fermentation products acetone and butanol. <br>Here we present a comprehensive comparison of proteome and transcriptome <br>data of continuously growing cells of C. acetobutylicum in a chemostat culture <br>under phosphate limitation at pH 5.7 (acidogenesis) and pH 4.5 (solventogenesis).
Project description:In this study the transcriptional behavior of the natural solvent producing bacterium Clostridium acetobutylicum was investigated following n butanol stress using DNA microarray analysis. Therefore, a phosphate-limited chemostat culture was established and n-butanol stress (0.9%) was added to acidogenic cells at pH 5.7.
Project description:Artificial electron carriers have been widely used to shift the solvent ratio towards butanol in acetone-butanol-ethanol (ABE) fermentation of solventogenic clostridia according to decreased hydrogen production. In this study, first insights on the molecular level were gained to explore the effect of methyl viologen addition to cultures of Clostridium acetobutylicum. Employing batch fermentation in mineral salts medium, the butanol:acetone ratio was successively increased from 2.3 to 12.4 on a 100 ml scale in serum bottles and from 1.4 to 16.5 on a 1,300 ml scale in bioreactors, respectively. The latter cultures were used for DNA microarray analyses to provide new information on the transcriptional changes referring to methyl viologen exposure and thus, exhibing gene expression patterns according to the manipulation of the cellular redox balance.
Project description:Clostridium acetobutylicum is a Gram-positive, endospore-forming bacterium that is considered as a strict anaerobe. It ferments sugars to the organic acids acetate and butyrate or shifts to formation of the solvents - ethanol, butanol and acetone. In most bacteria the major regulator of iron homeostasis is Fur (ferric uptake regulator). Analysis of the genome of Clostridium acetobutylicum has revealed three genes encoding Fur-like proteins. The amino acid sequece of one of them showed 70% similarity to the Fur protein of the closely related Bacillus subtilis.<br>Thus, to gain insight into the role of Fur and the mechanisms for maintenance of iron homeostasis in this strict anaerobic organism, we determined its transcriptional profile in response to iron limitation and inactivation of fur.
Project description:Clostridium acetobutylicum was grown in a batch-culture with minimal medium containing glucose and xylose as substrate. Diauxie growth was observed after glucose was consumed. Following the organism grows on xylose. Transcriptional analysis was done to pursue the cellular processes during the switch from growth on glucose to growth on xylose. We compared DNA-Microarray data from cells grown during the exponential phase on glucose (A), with cells growing during the start of diauxie growth lag (B), during the end of diauxie growth lag (C) and during exponential growth on xylose (D). We used cells grown in a continuous culture with glucose as substrate as common reference for the samples A-D.
Project description:Clostridium acetobutylicum is well-known for its butanol production. Butanol toxicity is a major drawback for the generation of high-butanol producing strains. Here, the transcriptional response a steady state, acidogenic (pH 6), phosphate-limited Clostridium acetobutylicum chemostat culture to different levels of n-butanol (0.25-1%) was investigated. For the butanol challenge experiments butanol (1-butanol) was added (a) to the supplying medium and (b) to the culture vessel to guarantee an immediate change in the butanol concentration. Addition of butanol to the culture was timed to match the supply of the new medium through the feedlines. The butanol concentration was increased stepwise in intervals of 66.6 h (5 volume changes) to moderate butanol concentrations of 0.25%, 0.5%, 0.75% and 1% (v/v).
Project description:In an effort to begin to delineate the bicarbonate regulon, we used microarray analysis with cells grown to late exponential growth phase (3 hr) and then submitted to a 15 min exposure with 0.1 M NaHCO3. Our goal was to define the first set of genes affected by the presence of bicarbonate.
Project description:Many plant researchers have applied genomic tools to model species to identify abiotic stress responsive genes that might be useful for improving stress tolerance in crops. However, it is unclear whether this translational approach will be successful given the complexity of abiotic stress tolerance. We carried out a functional genomic (ionomic, transcriptomic and metabolomic) comparison of three model and three forage species of the genus Lotus with varying tolerance to salinity. Transcriptome analysis showed that about 60 % of expressed genes were responsive to salt treatment in one or more of the six species tested, but less than 1 % was responsive in all genotypes. Therefore, genotype-specific responses overshadowed conserved transcriptional responses to salinity and represent an impediment to translational genomics. Fortunately, 'triangulation' from multiple species enabled the identification of a core set of conserved and tolerant-specific responses that could provide durable tolerance across genotypes.
Project description:L. japonicus (Regel) Larsen cv, Gifu were initially obtained from Prof. Jens Stougaard<br>(University Aarhus, Denmark) and then self-propagated at the University of Seville.<br>Seeds were scarified and surface-sterilized,<br>germinated in 1% agar Petri dishes, and transferred to pots using a 1:1 (v/v) mixture<br>of vermiculite and sand as solid support.<br>Five seedlings were planted in each pot and grown<br>during 35 days in a growth chamber under 16/8 hours<br>day/night, 20/18ï¿½C, with a photosynthetic photon flux<br>density of 250 ï¿½mol/m2ï¿½s and a constant humidity of 70%.<br> Plants were watered with Hornum nutrient solution.<br>Drought was applied withholding irrigation for the reported period<br>of time and sample plants or leaves were harvested for further analysis.