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Fermentation system
For this study, we used the DASbox Mini Bioreactor
System for microbial fermentation applications by
Eppendorf. Additionally, we connected a GA4 exhaust
gas analyzer unit including humidity sensors (Figure 1).
The GA4 measures the two off-gas components
oxygen (O2
) and carbon dioxide (CO2
The used GA4 version measures the O2
between 1 % and 50 % using a zirconium
dioxide sensor and CO2
is measured by a two-beam
infrared sensor. Together with the installed mass flow
sensor and humidity sensor, they allow for the online
calculation and tracking of the metabolic data of the
fermentation (OTR, CTR and RQ). For more detailed
information please refer to the GA4 manual, and the
Eppendorf Short Protocol 48. Several compensation
methods are available for humidity, volume, and
pressure compensation. To prevent liquid or foam
from entering the GA4 module and harming the
sensitive sensors we installed additional 250 mL
Schott bottles as foam traps between the gas outlet
of each DASbox mini bioreactor and the GA4.
Medium Preparation
We prepared three types of media for different
applications in this study (Mattanovich et al., 2012):
> One complex medium for inoculum preparation
in shake flasks (Table 1)
> One defined minimal medium for the batch
stage in the DASbox Mini Bioreactor System
(Table 2 and 3)
> One defined feeding medium for the fed-batch
stage (Table 4)
Medium for inoculum preparation
YNB-medium is prepared by fully dissolving the
components per liter DI water (Table 1) and sterile
filtration using a Ø 0.22 μm sterile filter.
PTM1 trace elements solution was filter sterilized
using a 0.22 μm filter.
We transferred 95 mL BSM medium to the DASbox
mini bioreactors and individually added 0.03 mL
Antifoam 204 (Sigma-Aldrich®, A6426) to each vessel,
which corresponds to an amount of 0.03 % (v/v).
Next, we autoclaved the vessels using a Systec
VE-95 autoclave. After cooling to room temperature,
the pH of the growth medium was adjusted to 5.0
GENengnews.com | 15
Media for batch fermentation
Basal salts medium (BSM) containing per liter (Table 2)
and PTM1 trace elements solution containing per liter
(Table 3):
https://www.eppendorf.com/fileadmin/knowledgebase/asset/OC-en/987404.pdf http://www.GENengnews.com


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