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Open Access Methodology

Development of a modified Respiration Activity Monitoring System for accurate and highly resolved measurement of respiration activity in shake flask fermentations

Sven Hansen, Ioanna Hariskos, Bettina Luchterhand and Jochen Büchs*

Author Affiliations

AVT. Biochemical Engineering, RWTH Aachen University, Worringerweg 1, Aachen 52074, Germany

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Journal of Biological Engineering 2012, 6:11  doi:10.1186/1754-1611-6-11

Published: 17 August 2012

Abstract

Background

The Respiration Activity Monitoring System (RAMOS) is an established device to measure on-line the oxygen transfer rate (OTR), thereby, yielding relevant information about metabolic activities of microorganisms and cells during shake flask fermentations. For very fast-growing microbes, however, the RAMOS technique provides too few data points for the OTR. Thus, this current study presents a new model based evaluation method for generating much more data points to enhance the information content and the precision of OTR measurements.

Results

In cultivations with E.coli BL21 pRSET eYFP-IL6, short diauxic and even triauxic metabolic activities were detected with much more detail compared to the conventional evaluation method. The decline of the OTR during the stop phases during oxygen limitations, which occur when the inlet and outlet valves of the RAMOS flask were closed for calibrating the oxygen sensor, were also detected. These declines reflected a reduced oxygen transfer due to the stop phases. In contrast to the conventional calculation method the new method was almost independent from the number of stop phases chosen in the experiments.

Conclusions

This new model based evaluation method unveils new peaks of metabolic activity which otherwise would not have been resolved by the conventional RAMOS evaluation method. The new method yields substantially more OTR data points, thereby, enhancing the information content and the precision of the OTR measurements. Furthermore, oxygen limitations can be detected by a decrease of the OTR during the stop phases.

Keywords:
Oxygen transfer rate; RAMOS; Shake flask; Bioprocess monitoring; Diauxic growth; Oxygen limitation