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Monosaccharide precursors for boosting chondroitin-like capsular polysaccharide production

  • Applied Microbial and Cell Physiology
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Abstract

Chondroitin sulfate is a well-known bioactive molecule, widely used as an anti-osteoarthritis drug, that is nowadays mainly produced by animal tissue sources with unsafe extraction procedures. Recent studies have explored an integrated biotechnological–chemical strategy to obtain a chondroitin sulfate precursor from Escherichia coli K4 capsular polysaccharide, demonstrating the influence of environmental and growth conditions on capsule synthesis. In this research work, the flexibility of the strain biosynthetic machinery was investigated to enhance the K4 capsular polysaccharide production by supplementing the growth medium with the monosaccharides (glucuronic acid, galactosamine and fructose) that constitute the chain. Shake flask experiments were performed by adding the sugars singularly or together, by testing monosaccharide different concentrations and times of addition and by observing the bacterial sugar consumption. A K4 capsular polysaccharide production enhancement, compared to the control, was observed in all cases of supplementation and, in particular, significant 68 and 57 % increases were observed when adding 0.385 mM glucuronic acid plus galactosamine or 0.385 mM fructose, respectively. Increased expression levels of the gene kfoC, coding for a K4 polymerase, evaluated in different growth conditions, confirmed the results at the molecular level. Furthermore, batch fermentations, performed in lab-scale reactors (2 L), allowed to double the K4 capsular polysaccharide production values obtained in shake flask conditions, by means of a strict control of the growth parameters.

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Acknowledgments

This research was supported by Ministero dell’ Istruzione, dell’Università e della Ricerca (MIUR), L297 project: “Produzione biotecnologica di condroitina”.

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Authors and Affiliations

  1. Centro Servizi di Ateneo “Grandi Apparecchiature”, Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy

    Odile Francesca Restaino, Irene di Lauro & Elisabetta Carlino

  2. Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy

    Donatella Cimini, Mario De Rosa & Chiara Schiraldi

Authors
  1. Odile Francesca Restaino
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  2. Irene di Lauro
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  3. Donatella Cimini
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  4. Elisabetta Carlino
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  5. Mario De Rosa
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  6. Chiara Schiraldi
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Correspondence to Chiara Schiraldi.

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Restaino, O.F., di Lauro, I., Cimini, D. et al. Monosaccharide precursors for boosting chondroitin-like capsular polysaccharide production. Appl Microbiol Biotechnol 97, 1699–1709 (2013). https://doi.org/10.1007/s00253-012-4343-2

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  • DOI: https://doi.org/10.1007/s00253-012-4343-2

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