Details
| Original language | English |
|---|---|
| Article number | e00562 |
| Journal | Biotechnology Reports |
| Volume | 28 |
| Early online date | 18 Nov 2020 |
| Publication status | Published - Dec 2020 |
Abstract
Polysialic acid (polySia) are α2,8- and/or α2,9-linked homopolymers with interesting properties for meningococcal vaccine development or the cure of human neurodegenerative disorders. With the goal to avoid large scale production of pathogenic bacteria, we compare in the current study the efficacy of conventional polySia production to recombinant approaches using the engineered laboratory safety strain E. coli BL21. High cell density cultivation (HCDC) experiments were performed in two different bioreactor systems. Increased cell densities of up to 11.3 (±0.4) g/L and polySia concentrations of up to 774 (±18) mg/L were reached in E. coli K1. However, cultivation of engineered E. coli BL21 strains delivered comparable cell densities but a maximum of only 133 mg/L polySia. Using established downstream procedures, host cell DNA and proteins were removed. All recombinant polySia products showed an identical degree of polymerization >90. Polymers with different glycosidic linkages could be successfully differentiated by nuclear magnetic resonance spectroscopy.
Keywords
- Escherichia coli, High cell density, NMR spectroscopy, Polysialic acid
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
Sustainable Development Goals
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In: Biotechnology Reports, Vol. 28, e00562, 12.2020.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Accelerated production of α2,8- and α2,9-linked polysialic acid in recombinant Escherichia coli using high cell density cultivation
AU - Bartling, Bastian
AU - Brüchle, Nora C.
AU - Rehfeld, Johanna S.
AU - Boßmann, Daniel
AU - Fiebig, Timm
AU - Litschko, Christa
AU - Fohrer, Jörg
AU - Gerardy-Schahn, Rita
AU - Scheper, Thomas
AU - Beutel, Sascha
N1 - Funding Information: This work was financially supported by the German Federal Ministry of Education and Research (BMBF-VIP+, grant numbers: BMBF-03VP00271 and BMBF-03VP00273). Furthermore, we would like to thank Monika Berger (Institute of Clinical Chemistry) for excellent technical assistance.
PY - 2020/12
Y1 - 2020/12
N2 - Polysialic acid (polySia) are α2,8- and/or α2,9-linked homopolymers with interesting properties for meningococcal vaccine development or the cure of human neurodegenerative disorders. With the goal to avoid large scale production of pathogenic bacteria, we compare in the current study the efficacy of conventional polySia production to recombinant approaches using the engineered laboratory safety strain E. coli BL21. High cell density cultivation (HCDC) experiments were performed in two different bioreactor systems. Increased cell densities of up to 11.3 (±0.4) g/L and polySia concentrations of up to 774 (±18) mg/L were reached in E. coli K1. However, cultivation of engineered E. coli BL21 strains delivered comparable cell densities but a maximum of only 133 mg/L polySia. Using established downstream procedures, host cell DNA and proteins were removed. All recombinant polySia products showed an identical degree of polymerization >90. Polymers with different glycosidic linkages could be successfully differentiated by nuclear magnetic resonance spectroscopy.
AB - Polysialic acid (polySia) are α2,8- and/or α2,9-linked homopolymers with interesting properties for meningococcal vaccine development or the cure of human neurodegenerative disorders. With the goal to avoid large scale production of pathogenic bacteria, we compare in the current study the efficacy of conventional polySia production to recombinant approaches using the engineered laboratory safety strain E. coli BL21. High cell density cultivation (HCDC) experiments were performed in two different bioreactor systems. Increased cell densities of up to 11.3 (±0.4) g/L and polySia concentrations of up to 774 (±18) mg/L were reached in E. coli K1. However, cultivation of engineered E. coli BL21 strains delivered comparable cell densities but a maximum of only 133 mg/L polySia. Using established downstream procedures, host cell DNA and proteins were removed. All recombinant polySia products showed an identical degree of polymerization >90. Polymers with different glycosidic linkages could be successfully differentiated by nuclear magnetic resonance spectroscopy.
KW - Escherichia coli
KW - High cell density
KW - NMR spectroscopy
KW - Polysialic acid
UR - http://www.scopus.com/inward/record.url?scp=85097002512&partnerID=8YFLogxK
U2 - 10.1016/j.btre.2020.e00562
DO - 10.1016/j.btre.2020.e00562
M3 - Article
C2 - 33304838
AN - SCOPUS:85097002512
VL - 28
JO - Biotechnology Reports
JF - Biotechnology Reports
M1 - e00562
ER -