Open AccessPolysialic acid production using Escherichia coli K1 in a disposable bag reactor
Author(s) -
Vries Ingo,
Busse Christoph,
Kopatz Jens,
Neumann Harald,
Beutel Sascha,
Scheper Thomas
Publication year - 2017
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.201600220
Subject(s) - polysialic acid , escherichia coli , chemistry , continuous stirred tank reactor , mixing (physics) , bioreactor , biochemistry , chromatography , monomer , polymerization , downstream processing , chemical engineering , food science , cell , organic chemistry , polymer , quantum mechanics , gene , engineering , neural cell adhesion molecule , cell adhesion , physics
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 mg protein /g polySia and a maximal chain length of ∼90 degree of polymerization.