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 -