Details
| Original language | English |
|---|---|
| Pages (from-to) | 723-731 |
| Number of pages | 9 |
| Journal | Engineering in life sciences |
| Volume | 17 |
| Issue number | 7 |
| Publication status | Published - 19 Jan 2017 |
Abstract
Polysialic acid (polySia), consisting of α-(2,8)-linked N-acetylneuraminic acid monomers plays a crucial role in many biological processes. This study presents a novel process for the production of endogenous polySia using Escherichia coli K1 in a disposable bag reactor with wave-induced mixing. Disposable bag reactors provide easy and fast production in terms of regulatory requirements as GMP, flexibility, and can easily be adjusted to larger production capacities not only by scale up but also by parallelization. Due to the poor oxygen transfer rate compared to a stirred tank reactor, pure oxygen was added during the cultivation to avoid oxygen limitation. During the exponential growth phase the growth rate was 0.61 h−1. Investigation of stress-related product release from the cell surface showed no significant differences between the disposable bag reactor with wave-induced mixing and the stirred tank reactor. After batch cultivation a cell dry weight of 6.8 g L−1 and a polySia concentration of 245 mg L−1 were reached. The total protein concentration in the supernatant was 132 mg L−1. After efficient and time-saving downstream processing characterization of the final product showed a protein content of below 0.04 mgprotein/gpolySia and a maximal chain length of ∼90 degree of polymerization.
Keywords
- Disposable bag reactor, Escherichia coli K1, Oxygen limitation, Polysialic acid, Stirred tank reactor
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
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In: Engineering in life sciences, Vol. 17, No. 7, 19.01.2017, p. 723-731.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Polysialic acid production using Escherichia coli K1 in a disposable bag reactor
AU - de Vries, Ingo
AU - Busse, Christoph
AU - Kopatz, Jens
AU - Neumann, Harald
AU - Beutel, Sascha
AU - Scheper, Thomas
N1 - Funding information: This work was financially supported by the German Research Foundation (DFG, Funding numbers: DFG-NE507/14-1 and DFG-SCHE279/35-1) and the German Federal Ministry of Education and Research (BMBF, Funding numbers: BMBF-03VP00271 and BMBF-03VP00273). We acknowledge all associates for helpful discussions and technical support. Drs. Jens Kopatz and Harald Neumann are named inventors on patent applications related to the use of polysialic acid for neurodegenerative diseases filed by the Universities of Bonn and Cologne.
PY - 2017/1/19
Y1 - 2017/1/19
N2 - Polysialic acid (polySia), consisting of α-(2,8)-linked N-acetylneuraminic acid monomers plays a crucial role in many biological processes. This study presents a novel process for the production of endogenous polySia using Escherichia coli K1 in a disposable bag reactor with wave-induced mixing. Disposable bag reactors provide easy and fast production in terms of regulatory requirements as GMP, flexibility, and can easily be adjusted to larger production capacities not only by scale up but also by parallelization. Due to the poor oxygen transfer rate compared to a stirred tank reactor, pure oxygen was added during the cultivation to avoid oxygen limitation. During the exponential growth phase the growth rate was 0.61 h−1. Investigation of stress-related product release from the cell surface showed no significant differences between the disposable bag reactor with wave-induced mixing and the stirred tank reactor. After batch cultivation a cell dry weight of 6.8 g L−1 and a polySia concentration of 245 mg L−1 were reached. The total protein concentration in the supernatant was 132 mg L−1. After efficient and time-saving downstream processing characterization of the final product showed a protein content of below 0.04 mgprotein/gpolySia and a maximal chain length of ∼90 degree of polymerization.
AB - Polysialic acid (polySia), consisting of α-(2,8)-linked N-acetylneuraminic acid monomers plays a crucial role in many biological processes. This study presents a novel process for the production of endogenous polySia using Escherichia coli K1 in a disposable bag reactor with wave-induced mixing. Disposable bag reactors provide easy and fast production in terms of regulatory requirements as GMP, flexibility, and can easily be adjusted to larger production capacities not only by scale up but also by parallelization. Due to the poor oxygen transfer rate compared to a stirred tank reactor, pure oxygen was added during the cultivation to avoid oxygen limitation. During the exponential growth phase the growth rate was 0.61 h−1. Investigation of stress-related product release from the cell surface showed no significant differences between the disposable bag reactor with wave-induced mixing and the stirred tank reactor. After batch cultivation a cell dry weight of 6.8 g L−1 and a polySia concentration of 245 mg L−1 were reached. The total protein concentration in the supernatant was 132 mg L−1. After efficient and time-saving downstream processing characterization of the final product showed a protein content of below 0.04 mgprotein/gpolySia and a maximal chain length of ∼90 degree of polymerization.
KW - Disposable bag reactor
KW - Escherichia coli K1
KW - Oxygen limitation
KW - Polysialic acid
KW - Stirred tank reactor
UR - http://www.scopus.com/inward/record.url?scp=85012981962&partnerID=8YFLogxK
U2 - 10.1002/elsc.201600220
DO - 10.1002/elsc.201600220
M3 - Article
AN - SCOPUS:85012981962
VL - 17
SP - 723
EP - 731
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 7
ER -