Establishing recombinant production of pediocin PA-1 in Corynebacterium glutamicum
2021
Goldbeck, Oliver | Desef, Dominique N. | Ovchinnikov, Kirill V. | Perez-Garcia, Fernando | Christmann, Jens | Sinner, Peter | Crauwels, Peter | Weixler, Dominik | Cao, Peng | Becker, Judith | Kohlstedt, Michael | Kager, Julian | Eikmanns, Bernhard J. | Seibold, Gerd M. | Herwig, Christoph | Wittmann, Christoph | Bar, Nadav S. | Diep, Dzung B. | Riedel, Christian U.
Bacteriocins are antimicrobial peptides produced by bacteria to inhibit competitors in their natural environments. Some of these peptides have emerged as commercial food preservatives and, due to the rapid increase in antibiotic resistant bacteria, are also discussed as interesting alternatives to antibiotics for therapeutic purposes. Currently, commercial bacteriocins are produced exclusively with natural producer organisms on complex substrates and are sold as semi-purified preparations or crude fermentates. To allow clinical application, efficacy of production and purity of the product need to be improved. This can be achieved by shifting production to recombinant microorganisms.Here, we identify Corynebacterium glutamicum as a suitable production host for the bacteriocin pediocin PA-1. C. glutamicum CR099 shows resistance to high concentrations of pediocin PA-1 and the bacteriocin was not inactivated when spiked into growing cultures of this bacterium. Recombinant C. glutamicum expressing a synthetic pedACDCᵍˡ operon releases a compound that has potent antimicrobial activity against Listeria monocytogenes and Listeria innocua and matches size and mass:charge ratio of commercial pediocin PA-1. Fermentations in shake flasks and bioreactors suggest that low levels of dissolved oxygen are favorable for production of pediocin. Under these conditions, however, reduced activity of the TCA cycle resulted in decreased availability of the important pediocin precursor l-asparagine suggesting options for further improvement. Overall, we demonstrate that C. glutamicum is a suitable host for recombinant production of bacteriocins of the pediocin family.
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