Bioprocess Engineering 18 (1998) 7577 Springer-Verlag 1998

Continuous production of citric acid with recirculation

of the fermentation broth after product recovery
Part 2: Product recovery and recirculation of the fermentation broth
S. Wieczorek, H. Brauer

Abstract For the production of citric acid a four stage

process consisting of the fermentation stage, extraction
stage, reextraction stage and product stage has been tested.
The present paper discusses the results of the extraction
and the reextraction stage. Further a fermentation with
recirculation of the fermentation broth after the product
recovery is explained. The product recovery is achieved by
a liquidliquid extraction in a spray column.
List of symbols
M kg
mass
t
d
time
V m3
volume
Greek symbol
q kg/m3 concentration
Subscripts
CA citric acid
R reactor

1
Introduction
Citric acid is produced by the fungi Aspergillus niger in an
aerob fermenter. For the product recovery of citric acid the
fermentation medium has to be taken out of the reactor. In
the continuous process it would not be economical and
ecological to discharge the fermentation broth after
product recovery took place. Therefore a recirculation of
the fermentation broth is recommended. For this purpose
a four stage process was created. Part 1 of the article
dealt with the fermentation stage of the process especially the continuous fermentation. This part deals with
the product recovery and the recirculation of the fermentation broth.
2
Extraction stage
The citric acid which is produced by the fungi Aspergillus
niger in the fermentation stage is soluted in the fermenReceived: 4 February 1997
S. Wieczorek, H. Brauer
Technische Universitat Berlin, Institut fur Verfahrenstechnik,
Strae des 17. Juni 136, D-10623 Berlin, Germany
Correspondence to: H. Brauer

tation broth. To separate the aqueous phase from the

biomass a microltration modul is placed inside the reciprocating-jet-bioreactor. The aqueous phase is then
pumped to the extraction stage with a liquid-liquid extraction to separate the citric acid. Main part of the plant is
a spray column with a height of 2000 mm and a diametre
of 40 mm. A schematic view of the plant is given in Fig. 1.
The citric acid containing fermentation broth enters the
column at the top and the citric acid reduced broth is
withdrawn at the bottom. Further the aqueous phase
passes an activated carbon lter and is pumped back to the
fermentation stage as recycled fermentation broth. The
extractant is dispersed at the bottom and the droplets are
collected at the upper end of the column. From there the
loaded extractant is pumped to the reextraction stage to
give off the citric acid and to be regenerated. After regeneration the extractant can be used for the extraction
process.

2.1
Extractant
Tertiary amines are very efcient binders for organic acids. They form an amine-acid complex with the organic
acid. For the experiments tri-dodecylamine was choosen as
a tertiary amine. To improve the solubility of this amineacid complex in the extractant octanol as a polar component was added to the extractant. Low viscosity and low
density promote phase separation. Therefore cerosin was
given to the extractant. Experiments were carried out in
the spray column to nd out the best combination of the
three components for the extraction of citric acid from the
fermentation broth. For the study an articial fermentation broth with the following composition was used: citric
acid 40 kg/m3, fructose 40 kg/m3, glucose 40 kg/m3 and
KH2PO4 0.3 kg/m3. The results can be seen in Fig. 2. The
distribution coefcient is given for different compositions
of the extractant. The curves have basically the same
shape. The distribution coefcient increases with increasing amount of octanol because the amine-acid complex
has to be soluted in the extractant. If the percentage of
octanol gets higher than 35% the distribution coefcient
decreases. This is a result of the smaller percentage of
cerosin as dilutent in the extractant. The curve given for an
extractant with 40% tri-dodecylamine shows the smallest
values for the distribution coefcient because of the
smallest amount of cerosin in the extractant. The highest
distribution coefcient is obtained for the extractant with
30% of tri-dodecylamin. For the following experiments a

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Bioprocess Engineering 18 (1998)

76

Fig. 3. Concentration of citric acid versus fermentation time for

different treatments of the fermentation broth

Fig. 1. Schematic view of the extraction stage

Fig. 2. Distribution coefcient versus percentage of octanol in

the extractant for different percentages of tri-dodecylamine

composition of 35% octanol, 35% cerosin and 30% tridodecylamine was choosen.

2.2
Treatment of the recycled fermentation broth
The extractant has a low solubility in water and therefore
parts of it are transferred into the fermentation reactor
after the extraction process. To examine the inuence of
each component of the extractant on citric acid production
experiments in shaken asks were carried out. The resulting two phase system was mixed with a magnetic
stirrer to reach the equilibrium state and only the aqueous
phase was given to the shaken asks. The fermentations in
the shaken asks were started with an inoculum of Aspergillus niger. To see the inuence of the components on
citric acid production a standard fermentation without any
extractant components prepared. In Fig. 3 one can see the
citric acid concentration in the different asks as a function of fermentation time. The highest citric acid concentration of 22 kg/m3 was reached by the standard

fermentation. A slight decrease with an acid concentration

of about 20 kg/m3 could be observed in the case of the
cerosin addition. The tri-dodecylamine reduced the citric
acid concentration down to about 10 kg/m3. Nearly no
citric acid was produced by the fungi which were in contact with octanol. The results show that the citric acid
production in the fermentation stage is strongly inuenced
by the extractant. One possibility to clean fermentation
broth of the extractant components is the use of activated
carbon. The fermentation media with the different components of the extractant were cleaned with an activated
carbon lter and were used for fermentation. The results
can be seen in Fig. 3 as well. All the fermentations reached
nearly the same citric acid concentration and the results
are presented in one curve. The citric acid concentration is
nearly as high as the concentration for the standard fermentation. It can be concluded that the cleaning step with
activated carbon is successful. Experiments showed that
about 1% of the sugar in the fermentation broth was retained in the lter.

3
Reextraction stage
In the reextraction stage the citric acid is removed from
the extractant and the extractant is regenerated. The
loaded extractant is mixed with an aqueous solution of trisodiumcitrat. One or two sodium ions leave the tri-sodiumcitrat to displace the amine in the amine-acid complex.
The two phase system consists of the unloaded extractant
and a mixture of mono- and di-sodiumcitrat. They are
separated in a settler. The aqueous phase is mixed with
soda lye to form tri-sodiumcitrat. A part of it is used for
the reextraction and the other part is converted into
crystallin citric acid in the product stage. In the reextraction stage the citric acid concentration in the extractant
could be reduced from 34.5 kg/m3 to 0.6 kg/m3. To be
recycled into the extraction stage the extractant needs
further cleaning. The regeneration is done by mixing the
extractant with an aqueous solution of strong soda lye.
Then the two phases are separated in a settler and the
aqueous phase is discharged as waste. For an economic
process it is of importance that the extractant can be used
a lot of times without loosing its ability to extract citric

S. Wieczorek, H. Brauer: Continuous production of citric acid

ous fermentation lasts for 66 days and the relative mass of

produced citric acid is 232 kg/m3. The batchwise fermentation reaches the maximum of 53 kg/m3 after 13 days. If
one would do ve batchwise fermentations after each other
a total mass of citric acid of 253 kg/m3 after 65 days would
be obtained. This is marked as a point at the end of the
doted line. Up to the 55th day the continuous fermentation
is as good as the ve successive done batchwise fermentations. Only at the end of the continous fermentation the
productivity of the biomass is strongly reduced. During
the continuous process over 66 days about 150 dm3 of the
fermentation medium were recirculated. This is about
seven times of the reactor volume. The volume of waste in
form of used fermentation broth could be reduced by the
continuous fermentation with recirculation. At the end of
a batchwise and continuous fermentation the whole reacFig. 4. Concentration of citric acid versus the number of
tor volume of 20 dm3 is waste. This means ve successive
regenerations of the extractant
done batch fermentations lead to 100 dm3 of waste. The
continuous fermentation with three dilution processes of
acid. In laboratory experiments the extractant was loaded 16 dm3 each built up only 68 dm3 of waste. In addition the
and regenerated several times. The maximum citric acid batch fermentations need four more times intensive
concentration in the extractant was measured for each
preparations of the inoculum compared to the continuous
extraction process. The results can be seen in Fig. 4. The process.
fresh extractant can take a citric acid concentration of
34.5 kg/m3. Even after 30 regeneration processes the ex5
tractant can take the same amount of citric acid.

Conclusion
It could be shown that a continuous fermentation with
4
recirculation of the fermentation broth after product reContinuous fermentation with recirculation
The details of a continuous fermentation have been dis- covery was run for 55 days with very good results. The
cussed in part one of the article. To see the inuence of the extraction stage causes the problem that parts of the exrecirculation of the fermentation broth the fermentation tractant dissolve in the aqueous phase. The necessary
cleaning of the recirculated fermentation broth was
has to run for a long time. This can be achieved by the
fermentation process with dilution of the biosuspension achieved with activated carbon. A regeneration of the extractant could be done for more than 30 times.
which has been explained in part one. The dilution is
3
3
achieved by replacing 16 dm of the 20 dm biosuspension
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