Abstract
During dual-phase fermentations using Escherichia coli engineered for succinic acid production, the productivity and viable cell concentration decrease as the concentration of succinic acid increases. The effects of succinic acid on the fermentation kinetics, yield, and cell viability were investigated by resuspending cells in fresh media after selected fermentation times. The cellular succinic acid productivity could be restored, but cell viability continuously decreased throughout the fermentations by up to 80% and subsequently the volumetric productivity was reduced. Omitting complex nutrients in the resuspension media had no significant effect on cellular succinate productivity and yield, although the viable cell concentration and thus the volumetric productivity was reduced by approximately 20%. By resuspending the cells, the amount of succinate produced during a 100-h fermentation was increased by more than 60%. The results demonstrate that by product removal succinic acid productivity can be maintained at high levels for extended periods of time.
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Werpy T, Petersen G (2004) Top value added chemicals from biomass, volume I. US Department of Energy, http://www1.eere.energy.gov/biomass/pdfs/35523.pdf
Zeikus JG, Jain MK, Elankovan P (1999) Biotechnology of succinic acid production and markets for derived industrial products. Appl Microbiol Biotechnol 51:545–552
Chatterjee R, Millard CS, Champion K, Clark DP, Donnelly MI (2001) Mutation of the ptsG gene results in increased production of succinate in fermentation of glucose by Escherichia coli. Appl Environ Microbiol 67:148–154
Lin H, Bennett GN, San KY (2005) Fed-batch culture of a metabolically engineered Escherichia coli strain designed for high-level succinate production and yield under aerobic conditions. Biotechnol Bioeng 90:775–779
Samuelov NS, Datta R, Jain MK, Zeikus JG (1999) Whey fermentation by Anaerobiospirillum succiniciproducens for production of a succinate-based animal feed additive. Appl Environ Microbiol 65:2260–2263
Kim DY, Yim SC, Lee PC, Lee WG, Lee SY, Chang HN (2004) Batch and continuous fermentation of succinic acid from wood hydrolysate by Mannheimia succiniciproducens MBEL55E. Enzyme Microb Technol 35:648–653
Agarwal L, Isar J, Meghwanshi GK, Saxena RK (2006) A cost effective fermentative production of succinic acid from cane molasses and corn steep liquor by Escherichia coli. J Appl Microbiol 100:1348–1354
Lee PC, Lee SY, Hong SH, Chang HN (2002) Isolation and characterization of a new succinic acid-producing bacterium, Mannheimia succiniciproducens MBEL55E, from bovine rumen. Appl Microbiol Biotechnol 58:663–668
Lin H, Bennett GN, San KY (2005) Chemostat culture characterization of Escherichia coli mutant strains metabolically engineered for aerobic succinate production: a study of the modified metabolic network based on metabolite profile, enzyme activity, and gene expression profile. Metab Eng 7:337–352
Donnelly MI, Millard CS, Clark DP, Chen MJ, Rathke JW (1998) A novel fermentation pathway in an Escherichia coli mutant producing succinic acid, acetic acid, and ethanol. Appl Biochem Biotechnol 70–2:187–198
Jantama K, Haupt MJ, Svoronos SA, Zhang XL, Moore JC, Shanmugam KT, Ingram LO (2008) Combining metabolic engineering and metabolic evolution to develop nonrecombinant strains of Escherichia coli C that produce succinate and malate. Biotechnol Bioeng 99:1140–1153
Vemuri GN, Eiteman MA, Altman E (2002) Effects of growth mode and pyruvate carboxylase on succinic acid production by metabolically engineered strains of Escherichia coli. Appl Environ Microbiol 68:1715–1727
Andersson C, Helmerius J, Hodge D, Berglund KA, Rova U (2009) Inhibition of succinic acid production in metabolically engineered Escherichia coli by neutralizing agent, organic acids, and osmolarity. Biotechnol Prog 25:116–123
Warnecke T, Gill RT (2005) Organic acid toxicity, tolerance, and production in Escherichia coli biorefining applications. Microb Cell Fact 4:25
Yedur S, Berglund KA, Dunuwila DD (2001) Succinic acid production and purification. U.S. Patent 6,265,190
Huh YS, Jun YS, Hong YK, Song H, Lee SY, Hong WH (2006) Effective purification of succinic acid from fermentation broth produced by Mannheimia succiniciproducens. Process Biochemistry 41:1461–1465
Schügerl K (2000) Integrated processing of biotechnology products. Biotechnol Adv 18:581–599
Cao N, Du J, Gong CS, Tsao GT (1996) Simultaneous production and recovery of fumaric acid from immobilized Rhizopus oryzae with a rotary biofilm contactor and an adsorption column. Appl Environ Microbiol 62:2926–2931
Meynial-Salles I, Dorotyn S, Soucaille P (2008) A new process for the continuous production of succinic acid from glucose at high yield, titer, and productivity. Biotechnol Bioeng 99:129–135
Sauer M, Porro D, Mattanovich D, Branduardi P (2008) Microbial production of organic acids: expanding the markets. Trends Biotechnol 26:100–108
Andersson C, Hodge D, Berglund KA, Rova U (2007) Effect of different carbon sources on the production of succinic acid using metabolically engineered Escherichia coli. Biotechnol Prog 23:381–388
Donnelly MI, Sanville-Millard CY, Nghiem NP (2004) Method to produce succinic acid from raw hydrolysates. U.S. Patent 6,743,610
Vemuri GN, Eiteman MA, Altman E (2002) Succinate production in dual-phase Escherichia coli fermentations depends on the time of transition from aerobic to anaerobic conditions. J Ind Microbiol Biotechnol 28:325–332
Lin H, Bennett GN, San KY (2005) Metabolic engineering of aerobic succinate production systems in Escherichia coli to improve process productivity and achieve the maximum theoretical succinate yield. Metab Eng 7:116–127
Lee SJ, Song H, Lee SY (2006) Genome-based metabolic engineering of Mannheimia succiniciproducens for succinic acid production. Appl Environ Microbiol 72:1939–1948
Sanchez AM, Bennett GN, San KY (2005) Novel pathway engineering design of the anaerobic central metabolic pathway in Escherichia coli to increase succinate yield and productivity. Metab Eng 7:229–239
Foster JW (2004) Escherichia coli acid resistance: tales of an amateur acidophile. Nat Rev Microbiol 2:898–907
Lin SKC, Du C, Koutinas A, Wang R, Webb C (2008) Substrate and product inhibition kinetics in succinic acid production by Actinobacillus succinogenes. Biochem Eng J 41:128–135
Corona-González RI, Bories A, González-Álvarez V, Pelayo-Ortiz C (2008) Kinetic study of succinic acid production by Actinobacillus succinogenes ZT-130. Process Biochem 42:1047–1053
van Maris AJA, Winkler AA, Porro D, van Dijken JP, Pronk JT (2004) Homofermentative lactate production cannot sustain anaerobic growth of engineered Saccharomyces cerevisiae: possible consequence of energy-dependent lactate export. Appl Environ Microbiol 70:2898–2905
van Maris AJA, Konings WN, van Dijken JP, Pronk JT (2004) Microbial export of lactic and 3-hydroxypropanoic acid: implications for industrial fermentation processes. Metab Eng 6:245–255
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The authors acknowledge the financial support of Norrbottens forskningsråd (The Research Council of Norrbotten) and the Kempe Foundation.
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Andersson, C., Petrova, E., Berglund, K. et al. Maintaining high anaerobic succinic acid productivity by product removal. Bioprocess Biosyst Eng 33, 711–718 (2010). https://doi.org/10.1007/s00449-009-0393-y
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DOI: https://doi.org/10.1007/s00449-009-0393-y