Abstract
Acetate is a potential low-cost carbon source and can be used for microbial production of valuable chemicals in Escherichia coli. In this study, separate and simultaneous inactivation of the ptsG, ptsI, and ptsP genes involved in the phosphoenolpyruvate:carbohydrate phosphotransferase system was performed in E. coli and the effects on cell growth and acetate assimilation were evaluated. The mutant strain with double deletion of ptsG and ptsP exhibited faster acetate use than the other mutants. Inactivation of ptsI seriously reduced acetate consumption. This work provides a novel engineering target for improving acetate use rates.
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References
Bettenbrock K, Sauter T, Jahreis K, Kremling A, Lengeler JW, Gilles ED (2007) Correlation between growth rates, EIIACrr phosphorylation, and intracellular cyclic AMP levels in Escherichia coli K-12. J Bacteriol 189(19):6891–6900
Bhatia SK, Yang YH (2017) Microbial production of volatile fatty acids: current status and future perspectives. Rev Environ Sci Biotechnol 16(6):1–19
Chen J, Li W, Zhang ZZ, Tan TW, Li ZJ (2018) Metabolic engineering of Escherichia coli for the synthesis of polyhydroxyalkanoates using acetate as a main carbon source. Microb Cell Fact 17(1):102
Cherepanov PP, Wackernagel W (1995) Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158(1):9–14
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(12):6640–6645
Gosset G (2005) Improvement of Escherichia coli production strains by modification of the phosphoenolpyruvate:sugar phosphotransferase system. Microb Cell Fact 4(1):14
Hara KY, Shimodate N, Ito M, Baba T, Mori H, Mori H (2009) Systematic genome-wide scanning for genes involved in ATP generation in Escherichia coli. Metab Eng 11(1):1–7
Hayden JD, Ades SE (2008) The extracytoplasmic stress factor, sigmaE, is required to maintain cell envelope integrity in Escherichia coli. PLoS One 3(2):e1573
Jahn S, Haverkorn van Rijsewijk BR, Sauer U, Bettenbrock K (2013) A role for EIIA(Ntr) in controlling fluxes in the central metabolism of E. coli K12. Biochim Biophys Acta 1833(12):2879–2889
Kuhlman TE, Cox EC (2010) Site-specific chromosomal integration of large synthetic constructs. Nucleic Acids Res 38(6):e92
Lee CR, Cho SH, Yoon MJ, Peterkofsky A, Seok YJ (2007) Escherichia coli enzyme IIANtr regulates the K+ transporter TrkA. Proc Natl Acad Sci USA 104(10):4124–4129
Li Y, Huang B, Wu H, Li Z, Ye Q, Zhang YP (2016) Production of succinate from acetate by metabolically engineered Escherichia coli. ACS Synth Biol 5(11):1299–1307
Liang Q, Zhang F, Li Y, Zhang X, Li J, Yang P, Qi Q (2015) Comparison of individual component deletions in a glucose-specific phosphotransferase system revealed their different applications. Sci Rep 5:13200
Luttmann D, Gopel Y, Gorke B (2012) The phosphotransferase protein EIIA(Ntr) modulates the phosphate starvation response through interaction with histidine kinase PhoR in Escherichia coli. Mol Microbiol 86(1):96–110
Ninfa AJ (2011) Unnecessary signaling: poorly named? J Bacteriol 193(18):4571–4573
Noh MH, Lim HG, Woo SH, Song J, Jung GY (2018) Production of itaconic acid from acetate by engineering acid-tolerant Escherichia coli W. Biotechnol Bioeng 115(3):729–738
Rabus R, Reizer J, Paulsen I, Saier MH Jr (1999) Enzyme INtr from Escherichia coli. A novel enzyme of the phosphoenolpyruvate-dependent phosphotransferase system exhibiting strict specificity for its phosphoryl acceptor, NPr. J Biol Chem 274(37):26185–26191
Sigala JC, Flores S, Flores N, Aguilar C, de Anda R, Gosset G, Bolivar F (2009) Acetate metabolism in Escherichia coli strains lacking phosphoenolpyruvate: carbohydrate phosphotransferase system; evidence of carbon recycling strategies and futile cycles. J Mol Microbiol Biotechnol 16(3–4):224–235
Wu QL, Guo WQ, Zheng HS, Luo HC, Feng XC, Yin RL, Ren NQ (2016) Enhancement of volatile fatty acid production by co-fermentation of food waste and excess sludge without pH control: the mechanism and microbial community analyses. Bioresour Technol 216:653–660
Xiao Y, Ruan Z, Liu Z, Wu SG, Varman AM, Liu Y, Tang YJ (2013) Engineering Escherichia coli to convert acetic acid to free fatty acids. Biochem Eng J 76(28):60–69
Xu X, Xie M, Zhao Q, Xian M, Liu H (2018) Microbial production of mevalonate by recombinant Escherichia coli using acetic acid as a carbon source. Bioengineered 9(1):116–123
Acknowledgements
This work was financially supported by the National Natural Science Foundation of China (31600066, 31100088, 31870105), the Shandong Provincial Natural Science Foundation (ZR2016CB20, ZR2016CL02), and the State Key Laboratory of Microbial Technology Open Projects Fund (M2016-10).
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Gu, P., Niu, H., Fan, X. et al. Engineering of phosphoenolpyruvate: carbohydrate phosphotransferase system increased acetate assimilation in Escherichia coli. 3 Biotech 9, 77 (2019). https://doi.org/10.1007/s13205-019-1600-4
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DOI: https://doi.org/10.1007/s13205-019-1600-4