Electrooptical measurements for monitoring metabolite fluxes in acetone–butanol–ethanol fermentations

Anisotropy of electrical polarizability in Clostridium acetobutylicum cells during pH 5 controlled acetone butanol ethanol fermentations was observed. Cell length was determined from the electrooptical data. Mean length was determined as being 2.5 μm in the growth phase and 3.5 μm in the early stationary phase. Based on the obtained frequency dispersion of polarizability anisotropy (FDPA) in the range of 190 to 2,100 kHz, the switch from the acidogenic to the solventogenic phase could be monitored. The slope of polarizability versus the frequency made it possible to differentiate between phases of dominating acid and solvent production. Metabolite fluxes determined from concentration measurements correlated well to the polarizability. A partial least‐squares (PLS) model was established and validated by applying data from several fermentations. The root mean square error of calibration (RMSEC) was 0.09 for the acid fluxes and 0.11 for the solvent fluxes. The root mean square error of prediction (RMSEP) was 0.20 for acid fluxes and 0.24 for solvent fluxes. The ratio of polarizability at high and low frequencies correlated to the ongoing sporulation process. At ratios below 0.25, spore formation in the cells became visible under the microscope. The advantage of using electrooptical measurements is the ability to observe metabolite fluxes rather than concentrations, which provides useful information on productivity during a bioprocess. Biotechnol. Bioeng. 2008;99: 862–869. © 2007 Wiley Periodicals, Inc.

[1]  R. Pethig,et al.  Viability of Giardia intestinalis Cysts and Viability and Sporulation State of Cyclospora cayetanensis Oocysts Determined by Electrorotation , 2001, Applied and Environmental Microbiology.

[2]  D. T. Jones,et al.  Acetone-butanol fermentation revisited. , 1986, Microbiological reviews.

[3]  A. Angersbach,et al.  Electro-optical analysis of bacterial cells , 2006 .

[4]  Bernhard Lendl,et al.  Mid-infrared spectroscopy coupled to sequential injection analysis for the on-line monitoring of the acetone–butanol fermentation process , 2001 .

[5]  M. Blanco,et al.  Analytical monitoring of alcoholic fermentation using NIR spectroscopy. , 2004, Biotechnology and bioengineering.

[6]  R. V. D. van den Heuvel,et al.  Development of markers for product formation and cell cycle in batch cultivation of Clostridium acetobutylicum ATCC 824 , 1998, Applied Microbiology and Biotechnology.

[7]  Paul Geladi,et al.  Principal Component Analysis , 1987, Comprehensive Chemometrics.

[8]  S. Arnold,et al.  Use of at‐line and in‐situ near‐infrared spectroscopy to monitor biomass in an industrial fed‐batch Escherichia coli process , 2002, Biotechnology and bioengineering.

[9]  P. Soucaille,et al.  Regulation of Clostridium acetobutylicum metabolism as revealed by mixed-substrate steady-state continuous cultures: role of NADH/NAD ratio and ATP pool , 1994, Journal of bacteriology.

[10]  Eva R. Kashket,et al.  Intracellular Conditions Required for Initiation of Solvent Production by Clostridium acetobutylicum , 1986, Applied and environmental microbiology.

[11]  E. Papoutsakis,et al.  The genes for butanol and acetone formation in Clostridium acetobutylicum ATCC 824 reside on a large plasmid whose loss leads to degeneration of the strain , 1997, Journal of bacteriology.

[12]  B. Kowalski,et al.  Partial least-squares regression: a tutorial , 1986 .

[13]  Hans P. Blaschek,et al.  Effect of Butanol Challenge and Temperature on Lipid Composition and Membrane Fluidity of Butanol-Tolerant Clostridium acetobutylicum , 1987, Applied and environmental microbiology.

[14]  E. Papoutsakis,et al.  Stoichiometric modeling of Clostridium acetobutylicum fermentations with non-linear constraints. , 1999, Journal of biotechnology.

[15]  E. Papoutsakis,et al.  Thiolase from Clostridium acetobutylicum ATCC 824 and Its Role in the Synthesis of Acids and Solvents , 1988, Applied and environmental microbiology.

[16]  S. Junne,et al.  Transcriptional analysis of product‐concentration driven changes in cellular programs of recombinant Clostridium acetobutylicumstrains , 2003, Biotechnology and bioengineering.

[17]  P. Dürre Formation of Solvents in Clostridia , 2005 .

[18]  D. Ivnitski,et al.  Electrooptical analysis of the Escherichia coli-phage interaction. , 2004, Analytical biochemistry.

[19]  Nasib Qureshi,et al.  Continuous production of butanol from starch-based packing peanuts , 2003 .

[20]  L. Ornella,et al.  De novo fatty acid synthesis is required for establishment of cell type‐specific gene transcription during sporulation in Bacillus subtilis , 1998, Molecular microbiology.