Abstract
Individual deletions of acs and aceA genes in E. coli B (BL21) showed little difference in the metabolite accumulation patterns but deletion of the ackA gene alone or together with pta showed acetic acid gradually accumulated to 3.1 and 1.7 g/l, respectively, with a minimal extended lag in bacterial growth and a higher pyruvate formation. Single poxB deletion in E. coli B (BL21) or additional poxB deletion in the ackA-pta mutants did not change the acetate accumulation pattern. When the acetate production genes (ackA-pta-poxB) were deleted in E. coli B (BL21) acetate still accumulated. This may be an indication that perhaps acetate is not only a by-product of carbon metabolism; it is possible that acetate plays also a role in other cellular metabolite pathways. It is likely that there are alternative acetate production pathways.
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References
Abdel-Hamid AM, Attwood MM, Guest JR (2001) Pyruvate oxidase contributes to the aerobic growth efficiency of Escherichia coli. Microbiology 147:1483–1493
Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97:6640–6645
De May M, De Maeseneire S, Soetaert W et al (2007) Minimizing acetate formation in E. coli fermentations. J Ind Microbiol Biotechnol 34:689–700
Dittrich CR, Bennett GN, San KY (2005) Characterization of the acetate-producing pathways in Escherichia coli. Biotechnol Prog 21:1062–1067
El-Mansi M (2005) Free CoA-mediated regulation of intermediary and central metabolism: an hypothesis which accounts for the excretion of alpha-ketoglutarate during aerobic growth of Escherichia coli on acetate. Res Microbiol 156:874–879
Kang Z, Geng Y, Xia Y et al (2009) Engineering Escherichia coli for an efficient aerobic fermentation platform. J Biotechnol 144(1):58–63
Kumari S, Simel EJ, Wolfe AJ (2000) Sigma (70) is the principal sigma factor responsible for transcription of acs, which encodes acetyl coenzyme A synthetase in Escherichia coli. J Bacteriol 182:551–554
Lee SY (1996) High cell density culture of Escherichia coli. Trends Biotechnol 14:98–105
Miller JH (1992) A short course in bacterial genetics. A laboratory manual and handbook for Escherichia coli and related bacteria. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 268–272
Noronha SB, Yeh HJ, Spande TF et al (2000) Investigation of the TCA cycle and the glyoylate shunt in Escherichia coli BL21 and JM109 using 13C-NMR/MS. Biotechnol Bioeng 68:316–327
Phue JN, Noronha SB, Hattacharyya R et al (2005) Glucose metabolism at high density growth of E. coli B and E. coli K: differences in metabolic pathways are responsible for efficient glucose utilization in E. coli B as determined by microarrays and Northern blot analyses. Biotechnol Bioeng 90:805–820
Shin S, Chang DE, Pan JG (2009) Acetate consumption activity directly determines the level of acetate accumulation during Escherichia coli W3110 growth. J Microbiol Biotechnol 19(10):1127–1134
Yang Y-T, Aristidou AA, San K-Y et al (1999) Metabolic flux analysis of Escherichia coli deficient in the acetate production pathway and expressing the Bacillus subtilis acetolactate synthase. Metab Eng 1:26–34
Acknowledgments
Funding was provided by the Intramural program at the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health. The authors would like to thank D. Livnat for proof reading of the manuscript.
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Purpose of work
Compared with E. coli K-12, E. coli B (BL21) is low acetate producer. To identify new ways to decrease acetate in E. coli K-12, the effect of deleting genes involved in the production and consumption of acetate in E. coli B (BL21) was evaluated.
Je-Nie Phue, Sang Jun Lee, Equal contribution with 1st author.
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Phue, JN., Lee, S.J., Kaufman, J.B. et al. Acetate accumulation through alternative metabolic pathways in ackA − pta − poxB − triple mutant in E. coli B (BL21). Biotechnol Lett 32, 1897–1903 (2010). https://doi.org/10.1007/s10529-010-0369-7
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DOI: https://doi.org/10.1007/s10529-010-0369-7