Abstract
Forty-eight single-use stirred tank bioreactors on a 10-mL scale operated in a magnetically inductive driven bioreaction block and automated with a liquid handler were applied for discrimination of different riboflavin producing Bacillus subtilis strains based on their performances in the parallel fed-batch processes. It was shown that a discrimination of the B. subtilis riboflavin producer strains can efficiently be achieved within one parallel fermentation run based on the integral riboflavin yield after 48 h. The possibility to perform replicates within the parallel fermentation run allows for a robust statistical analysis and is a prerequisite for the discrimination of producer strains under fed-batch process conditions. Within the estimation error, all of the riboflavin producing B. subtilis strains under study showed the same fed-batch process performances on the litre scale compared to the millilitre scale.
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References
Betts J, Doig S, Baganz F (2006) Characterization and application of a miniature 10 mL stirred-tank bioreactor, showing scale-down equivalence with a conventional 7 L reactor. Biotechnol Prog 22:681–688
Harms P, Kostov Y, French J, Soliman M, Anjanappa M, Ram A, Rao G (2006) Design and performance of a 24-station high throughput microbioreactor. Biotechnol Bioeng 93:6–13
Hümbelin M, Griesser V, Keller T, Schurter W, Haiker M, Hohmann H-P, Ritz H, Richter G, Bacher A, van Loon APGM (1999) GTP cyclohydrolase II and 3, 4-dihydroxy-2-butanone 4-phosphate synthase are rate-limiting enzymes in riboflavin synthesis of an industrial Bacillus subtilis strain used for riboflavin production. J Indust Microbiol Biotechnol 22:1–7
Knorr B, Schlieker H, Hohmann H-P, Weuster-Botz D (2007) Scale-down and parallel operation of the riboflavin production process with Bacillus subtilis. Biochem Eng J 33:263–274
Kostov Y, Harms P, Randers-Eichhorn L, Rao G (2001) Low-cost microbioreactor for high-throughput bioprocessing. Biotechnol Bioeng 72:346–352
Kusterer A, Krause C, Kaufmann K, Arnold M, Weuster-Botz D (2008) Fully automated single-use stirred-tank bioreactors for parallel microbial cultivations. Bioproc Biosyst Eng 31:207–215
Lamping SR, Zhang H, Allen B, Shamlou PA (2003) Design of a prototype miniature bioreactor for high throughput automated bioprocessing. Chem Eng Sci 58:747–758
Lehmann M (2007): Modified transketolase and use thereof. European patent number WO 2007/051552 A1).
Link H, Weuster-Botz D (2006) Genetic algorithm for multi-objective experimental optimization. Bioproc Biosyst Eng 29:385–390
Perkins J, Sloma A, Hermann T, Thieriault K, Zachgo E, Erdenberger T, Hannet N, Chatterjee N, Williams V, Rufo G, Hatch R, Pero J (1999) Genetic engineering of Bacillus subtilis for the commercial production of riboflavin. J Indust Microbiol Biotechnol 22:8–12
Puskeiler R, Kaufmann K, Weuster-Botz D (2005a) Development, parallelization and automation of a gas-inducing milliliter-scale bioreactor for high-throughput bioprocess design (HTBD). Biotechnol Bioeng 89:512–523
Puskeiler R, Kusterer A, John G, Weuster-Botz D (2005b) Miniature bioreactors for automated high-throughput bioprocess design (HTBD): Reproducibility of parallel fed-batch cultivations with Escherichia coli. Biotechnol Appl Biochem 42:227–235
Rudolf M, Kulisch W (2008) Biostatistik. Pearson Studium, München
Vallejos J, Kostov Y, Ram A, French J, Marten M, Rao G (2006) Optical analysis of liquid mixing in a minibioreactor. Biotechnol Bioeng 93:906–911
Weuster-Botz D, Puskeiler R, Kusterer A, Kaufmann K, John G, Arnold M (2005) Methods and milliliter scale devices for high-throughput bioprocess design. Bioproc Biosyst Eng 28:109–119
Acknowledgements
The authors gratefully acknowledge the technical assistance of Eva Chervenkova, Fei Xue and Katharina Kinast in the labs at the Technische Universität München and the funding of the research work by DSM Nutritional Products, Basel, Switzerland.
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Vester, A., Hans, M., Hohmann, HP. et al. Discrimination of riboflavin producing Bacillus subtilis strains based on their fed-batch process performances on a millilitre scale. Appl Microbiol Biotechnol 84, 71–76 (2009). https://doi.org/10.1007/s00253-009-1966-z
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DOI: https://doi.org/10.1007/s00253-009-1966-z