PECULIARITIES OF THE CULTIVATION OF ESCHERICHIA COLI BACTERIA DURING LEVOCARNITINE BIOSYNTHESIS
DOI:
https://doi.org/10.31073/foodresources2023-21-15Keywords:
cultivation, Escherichia coli, Levocarnitine, nutrient medium, nitrogen, glucose, promoter activityAbstract
Subject. Levocarnitine is a replaceable amino acid that performs an important function in the human body. The increase in commercial demand for L-carnitine has led to a surge in research on its biosynthesis using various strains of microorganisms. Because of the transience of cultivation conditions, it is quite difficult to study the laws of growth in dynamically changing environments. Purpose. Study the main properties of Escherichia coli bacteria when cultivated on different nutrient media. Methods. Bacterial growth rate, promoter activity, and substrate level were measured by robotic analysis with high temporal resolution throughout the growth curve in batch culture. Results. Cultivation of Escherichia coli for levocarnitine biosynthesis was carried out on media. Active biomass growth on McConkey Agar was observed after 72 hours of cultivation, while on bile agar biomass growth was less than 50%. Levocarnitine biosynthesis processes were studied at different concentrations of nitrogen nutrients in the form of NH4Cl or glucose carbon in the media. It was found that at low nitrogen concentrations (0,24 mM), cells suddenly stop growing, going from maximum to zero growth within 27 minutes. At the highest nitrogen concentration of 18,7 mM, when nitrogen is not limiting, cells gradually slow down and enter a stationary phase in about 4 hours. At intermediate nitrogen concentrations, cells first gradually slow down growth, and then suddenly stop it, which corresponds to Mono's law. Similar results were obtained using glucose as a limiting factor. The study of the promoter activity of various genes revealed that a sharp stoppage of growth is accompanied by a pulse regulation of gene expression in the corresponding pathways of assimilation of nutrients. Promoter activity of nitrogen and carbon assimilation genes increases sharply in the last generation before growth ceases. Scope of results. The research results will be used to establish the conditions for the cultivation of microorganisms under a limited concentration of substrate nutrients.
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Alhasaniah A. H. (2023). l-carnitine: Nutrition, pathology, and health benefits. Saudi J Biol Sci. 2023 Feb;30(2):103555. https://doi.org/10.1016/j.sjbs.2022.103555. Epub 2022 Dec 30. PMID: 36632072; PMCID: PMC9827390.
Fielding, R., Riede, L., Lugo, J. P., Bellamine, A. (2018). l-Carnitine Supplementation in Recovery after Exercise. Nutrients. Mar, 13;10(3):349. https://doi.org/10.3390/nu10030349. Erratum in: Nutrients. 2018 Apr 26;10(5): PMID: 29534031; PMCID: PMC5872767.
Jen-Hong, Chu. (2023). Effects of Various Lipid Sources and L-Carnitine Supplementation on the Growth Performance, Body Composition, and Antioxidants Enzyme Ability of Dog Conch Larvae, Laevistrombus canarium. Aquaculture Research. Vol. 2023, Article ID 5336474, 12 p. https://doi.org/10.1155/2023/5336474.
Britannica, The Editors of Encyclopaedia. (2020). "pure culture". Encyclopedia Britannica, 11 Feb. 2020, https://www.britannica.com/science/pure-culture. Accessed 21 September 2023.
Wanner U, Egli T: Dynamics of microbial growth and cell composition in batch culture. FEMS Microbiol Rev. 1990. № 6. P. 19–43.
Zaslaver, A., Bren, A., Ronen, M., Itzkovitz, S., Kikoin, I., Shavit, S., Liebermeister, W., Surette, M.G., Alon, U. (2006). A comprehensive library of fluorescent transcriptional reporters for Escherichia coli. Nat Methods. № 3. С. 623–628. https://doi.org/10.1038/nmeth895.
Bren, A., Hart, Y., Dekel, E. et al. (2013). The last generation of bacterial growth in limiting nutrient. BMC Syst Biol 7, 27 (2013). https://doi.org/10.1186/1752-0509-7-27.
Fink, J.W., Held, N.A., Manhart, M. (2023). Microbial population dynamics decouple growth response from environmental nutrient concentration. Proc Natl Acad Sci USA. 2023 Jan 10;120(2):e2207295120. https://doi.org/10.1073/pnas.2207295120. Epub 2023 Jan 4. PMID: 36598949; PMCID: PMC9926246.
Kim, M., Zhang, Z., Okano, H., Yan, D., Groisman, A., Hwa, T. (2012). Need-based activation of ammonium uptake in Escherichia coli. Mol Syst Biol. 2012;8:616. https://doi.org/10.1038/msb.2012.46. PMID: 23010999; PMCID: PMC3472687.
Jahn, S., Haverkorn van Rijsewijk, B .R., Sauer, U., Bettenbrock, K. (2013). A role for EIIA(Ntr) in controlling fluxes in the central metabolism of E. coli K12. Biochim Biophys Acta. 2013 Dec;1833(12):2879-2889. https://doi.org/10.1016/j.bbamcr.2013.07.011. Epub 2013 Jul 23. PMID: 23886628.
Ninfa, A. J., Magasanik, B. (1986). Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proceedings of the National Academy of Sciences. 83. P. 5909–5913. https://doi.org/10.1073/pnas.83.16.5909.
Serra, D. O., Hengge R. (2014). Stress responses go three dimensional – the spatial order of physiological differentiation in bacterial macrocolony biofilms. Environ Microbiol. 2014 Jun;16(6):1455-71. https://doi.org/10.1111/1462-2920.12483. Epub 2014 May 5. PMID: 24725389; PMCID: PMC4238805.
Hart Y, Madar D, Yuan J, Bren A, Mayo AE, Rabinowitz JD, Alon U. Robust control of nitrogen assimilation by a bifunctional enzyme in E. coli. Mol Cell. 2011.№ 41. Р. 117–127. https://doi.org/10.1016/j.molcel.2010.12.023.
Mo, R., Liu, Y., Chen, Y., Mao, Y., Gao, B. (2022). Evolutionary Principles of Bacterial Signaling Capacity and Complexity. mBio. 2022 Jun 28;13(3):e0076422. https://doi.org/10.1128/mbio.00764-22. Epub 2022 May 10. PMID: 35536007; PMCID: PMC9239204.
Serra, D. O., Hengge, R. (2019). A c-di-GMP-Based Switch Controls Local Heterogeneity of Extracellular Matrix Synthesis which Is Crucial for Integrity and Morphogenesis of Escherichia coli Macrocolony Biofilms. J Mol Biol. 2019 Nov 22;431(23):4775-4793. https://doi.org/10.1016/j.jmb.2019.04.001. Epub 2019 Apr 4. PMID: 30954572.
Iyer, M. S, Pal, A., Venkatesh, K. V. (2023). A Systems Biology Approach To Disentangle the Direct and Indirect Effects of Global Transcription Factors on Gene Expression in Escherichia coli. Microbiol Spectr. 2023 Feb 7;11(2):e0210122. https://doi.org/10.1128/spectrum.02101-22. Epub ahead of print. PMID: 36749045; PMCID: PMC10100776.
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