Scale down and High Throughput Technologies

Internal growth control for high density cultures

Scheme of the working principle of enzyme controlled substrate delivery

© Neubauer

New and innovative high cell density culture strategies for microbial shaken cultures are developed on the basis of automatic substrate delivery. Initial solutions have been developed with the Finnish company BioSilta Oy and the University of Oulu and are known as the Enbase® technology.
Enzyme controlled substrate delivery provides a tight growth control in shaken cultures and has been proven to be highly advantageous for recombinant protein production, library screening and plasmid DNA production.

Currently we exploit this new cultivation idea for new organisms and substrates and in the scale up of bioprocesses. Additionally there are novel cultivation strategies in the focus of our research work.


Involved researchers:

Peter Neubauer, Julia Glazyrina, Mirja Krause, Stefan Junne, Friederike Hillig


Cooperation partners:

Biosilta Oy, Oulu, Finland
BPEL, University of Oulu, Oulu, Finland

Cellution Biotech BV, Assen, The Netherlands
Sartorius Stedim Biotech SA, Göttingen, Germany


Recent Publications:

Glazyrina J, Krause M, Junne S, Glauche F, Strom D, Neubauer P. 2011. Glucose-limited high cell density cultivations from small to pilot plant scale using an enzyme-controlled glucose delivery system. New Biotechnology [epub ahead of print]

Pilarek M, Glazyrina J, Neubauer P. 2011. Enhanced growth and recombinant protein production of Escherichia coli by a perfluorinated oxygen carrier in miniaturized fed-batch cultures. Microbial Cell Factories 27 (10):50.

Glazyrina J, Materne EM, Dreher T, Storm D, Junne S, Adams T, Greller G, Neubauer P. 2010. High cell density cultivation and recombinant protein production with Escherichia coli in a rocking-motion-type bioreactor. Microbial Cell Factories 30 (9):42.

Siurkus J, Panula-Perälä J, Horn U, Kraft M, Rimseliene R, Neubauer P. 2010:
Novel approach of high cell density recombinant bioprocess development: optimisation and scale-up from microliter to pilot scales while maintaining the fed-batch cultivation mode of E. coli cultures. Microbial Cell Factories 20 (9):35.

Krause M, Ukkonen K, Haataja T, Ruottinen M, Glumoff T, Neubauer A, Neubauer P, Vasala A. 2010: A novel fed-batch based cultivation method provides high cell-density and improves yield of soluble recombinant proteins in shaken cultures. Microbial Cell Factories 19(9):11.

Panula-Perälä J., Siurkus J., Vasala A., Wilmanowski R., Casteleijn M.G., Neubauer P. 2008. Enzyme controlled glucose auto-delivery system for high cell density cultivations in microplates and shake flasks. Microb Cell Fact 7:31.

Impact of gradients in large scale bioprocesses on the amino acid synthesis in Escherichia coli

Scale down reactor for simulation of concentration gradients in large scale bioprocesses

© Neubauer

Due to insufficient mixing in large-scale (mostly industrial) fermentation, inhomogeneities emerge. The impacts of the inhomogeneities can only be observed in laboratory scale, if the large scale conditions are imitated. Therefore, a combination of a stirred tank reactor and a plug flow reactor is applied, which enables the distribution of the culture broth into two compartments were different conditions exist.

When the microorganisms flow through the plug flow reactor, they experience an imitation of the feeding zone with a high glucose concentration and anaerobiosis. With variations in reactor conditions, the effect of a wide spectrum of fermentation conditions on different microorganisms can be determined. With the according analysis systems the whole spread from genome to metabolome can be investigated and the impacts of bad mixing conditions can be quantified. Research is focused on the observation of the amino acid synthesis in Escherichia coli. Conditions as they appear in large scale bioprocesses can change the product range and quantities of amino acids which are produced, leading to unwanted byproducts.
Beside the observation of the genome and proteome, also metabolic flux analyses are performed to describe and locate the changes on the microorganism’s metabolism.


Involved researchers:

Eva Brand, Peter Neubauer, Stefan Junne

Cooperation partners:

AG Prof. Schweder, Universität Greifswald, Germany


Recent Publications:

1. Junne S, Klingner A, Kabisch J, Schweder T, Neubauer P. 2011.
A two-compartment bioreactor system made of commercial parts for bioprocess scale-down studies: impact of oscillations on Bacillus subtilis fed-batch cultivations. Biotechnology Journal 6 (8):1009-17.

2. Neubauer P, Junne S. 2010. Scale-down simulators for metabolic analysis of large-scale bioprocesses. Current Opinion on Biotechnology 21 (1):114-21.

3. Soini J., Falschlehner C., Liedert C., Bernhard J., Vuoristo J., Neubauer, P. 2008. Norvaline is accumulated after a down-shift of oxygen in Escherichia coli W3110. Microb Cell Fact. 7:30.

4. Soini, J., Ukkonen K., Neubauer, P. 2008. High cell density media for Escherichia coli are generally designed for aerobic cultivations – consequences for large-scale bioprocesses and shake flask cultures. Microbial Cell Factories 7:26.