Czech J. Food Sci. Vol. 22, No.

3: 87–94

Modern Method of Lactic Acid Recovery

from Fermentation Broth

V��� HÁBOVÁ, K���� MELZOCH and M����� RYCHTERA

Department of Fermentation Chemistry and Bioengineering, Institute of Chemical Technology

in Prague, Prague, Czech Republic

Abstract

H����� V., M������ K., R������� M. (2004): Modern method of lactic acid recovery from fermentation broth.
Czech J. Food Sci., 22: 87–94.

Electrodialysis was used for lactic acid recovery from fermentation broth. In the first step, lactate was recovered
and concentrated by desalting electrodialysis, and the second step was electroconversion of lactate to lactic acid
by water-splitting electrodialysis. The final lactic acid concentration of 151 g/l was obtained. Total energy required
in both electrodialysis processes was about 1.5 kWh per 1 kg of lactic acid obtained. The fermentation broth had
to be pretreated prior to the electrodialysis experiments. The pretreatment consisted of ultrafiltration, decolouri-
sation, and the removal of multivalent metal ions.

Keywords: lactic acid recovery; electrodialysis; pretreatment of fermentation broth

Lactic acid (2-hydroxypropionic acid) was Generally, lactic acid can by manufactured
discovered for the first time in 1780 by a Swed- by either biological fermentation or chemical
ish chemist Scheele and since 1881 it has been synthesis. While the chemical synthesis pro-
produced commercially by fermentation. Lactic duces a racemic mixture of lactic acid (�(+)- and
acid is frequently used in food industry, esp. for �(–)-forms), fermentation technology is able to
the beverage production and in pharmaceutical selectively synthesise the desired enantiomer due
industry, chemical industry, or medicine (V��� to the high stereospecifity of several lactic acid
R�� 1985). The recent growing interest for the bacteria. The traditional fermentation process is
manufacture of biodegradable plastics initiates carried out in the batch mode at a constant pH
a high demand for lactic acid as the raw mate- between 5.5 and 6.0 and at temperatures above
rial for polylactate production (D���� et al. 1995; 40°C, with molasses or other sugars as the C-source
S�������� & S���� 2002). (K����� et al. 1996).
Another very promising lactic acid application is There are a lot of studies on lactic acid fermen-
the production of environmentally friendly “green” tation, on the production of �(+) lactic acid, and
solvents (lactate esters). They can replace traditional on the use of the agricultural cellulosic feedstock
solvents made from petrochemical feedstocks (T��� and waste for this purpose (I��� & L�� 1999; B��
et al. 1999). et al. 2003a,b; J�� et al. 2003; W������ et al. 2003).

Supported from EUREKA ∑! 1820 BIOLACTATE and by the Ministry of Education, Youth and Sports of the Czech
Republic, Grant No. CEZ: J19/98: 223300005.

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Vol. 22, No. 3: 87–94 Czech J. Food Sci.

To obtain lactic acid of the required purity, efficient Lactobacillus plantarum L10 as the producent strain
downstream processing is necessary. The conven- (the strain was obtained from the Collection of
tional fermentation process produces calcium lactate Department of Dairy and Fat Technologies, ICT
precipitate which has to be reacidified with a strong Prague, Czech Republic). The lactate concentra-
acid; large amounts of effluents arise, in particular tion ranged from 17 to 88 g/l. The initial medium
gypsum. It was estimated that downstream processes consisted, respectively, of (g/l): glucose, 40–100;
absorb about 80% of the production costs, indicating yeast extract, 5.3; KH2PO4, 2.6; CH3COONa.3H2O,
an urgent need for more economical downstream 6.6; ammonium citrate 2.6; MgSO 4.7H 2O, 0.24;
technologies, and for bioengineers to design and MnSO4.4H2O. The fermentation temperature was
operate them (H���� 2004). 37°C and pH was kept at 5.8 by means of NaOH
Recently, many investigations were focused on (20 g/l).
the downstream processing of lactic acid from Electrodialysis equipment. The electrodialysis
fermentation broth with the aim to reduce costs, laboratory unit BEL-500 (Berghof, Germany)
to simplify the recovery procedure, to reduce the consisted of a control unit (adjustable outputs of
contents of effluents, and to decrease the negative voltage from 0 to 50 V and of current from 0 to
impact on the surrounding environment (T����� et 3.9 A), a measuring device (conductivity and volt-
al. 1994; M�������� et al. 1998; F������� & S������� age), and 3 independent circuits with pumps and
1999; C�� et al. 2002). storage containers (for the diluate, the concentrate,
Membrane techniques, mainly electrodialysis and the electrode solution). The membrane stack
and nanofiltration, have been shown as promis- ED 0 (Mega, Czech Republic) with 20 pairs of ion
ing ways for the lactic acid recovery (J������ et exchange membranes Ralex CMH and Ralex AMH
al. 1996; B����� 2002; C��� et al. 2002). L�� et al. was used for the desalting in the electrodialysis
(1998) used a two-stage electrodialysis method for experiments. The effective membrane area was
the lactic acid recovery and K�� and M��� (2001) 180 cm2, the distance between the membranes was
used one-stage electrodialysis with two- and three- 1 mm. Stack Type 500 with 4 bipolar membranes
compartment water-splitting electrodialysis. L� et Neosepta BP-1 and 5 cation exchange membranes
al. (2004) proposed an electrokinetic membrane Neosepta CMB (Tokuyama Corp., Japan) was used
bioreactor for the lactic acid production that couples for water-splitting electrodialysis, the distance
fermentation with in situ lactic acid separation and between membranes being 0.5 mm. The effective
the product concentration. B�������� et al. (1998) membrane area was 57.6 cm2.
studied an integrated process using electrodialysis Operating conditions. Electrodialysis experi-
for the lactic acid recovery. M��-T��� et al. (2003) ments were carried out in the batch mode.
described the production of �(+) lactic acid by the Desalting electrodialysis: The electrode solution
electrodialysis fermentation. (Na2SO4 – 25 g/l), the concentrate (sodium lactate
– initial concentration 3–104 g/l), and the diluate
MATERIAL AND METHODS (the pretreated fermentation broth – the initial
concentration from 15 to 88 g/l) were circulated
Chemicals. Sodium lactate (p. a. purity) was through the corresponding compartments of the
from Sigma, the other chemicals were chemi- desalting stack at the flow rate of 2.4 l/min. For
cally pure products of Lachema (Brno, Czech the constant current period, voltage of 1.5 V per
Republic). Demineralised water (resistivity of one pair and current density of 7.8 mA/cm2 were
18.2 MΩ.cm) was prepared from distilled water used. For the constant voltage period, voltage of
in the device Millipore – Q gradient (Molsheim, 18 V was used. The experiments were terminated
France). Granulated active charcoal Purolite AC when the lactate concentration in the diluate
20G (Purolite International Ltd., UK) was used for dropped to 1–2 g/l.
decolourisation of the fermentation broth. Chelat- In the case of the two-level electrodialysis, only
ing resin Purolite S 940 (Purolite International Ltd., the depleted diluate was replaced by another
UK) and the weak cation exchanger Duolite C-433 fresh fermentation broth after the first level. The
(Fluka, Switzerland) were used for the recovery conditions of the second level were the same as
of multivalent metal ions. mentioned above.
Fermentation broth. Real fermentation broth was For water-splitting electrodialysis, the follow-
obtained through lactic acid fermentation using ing solutions were used: NaOH – 20 g/l (electrode

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Czech J. Food Sci. Vol. 22, No. 3: 87–94

solution), NaOH – 1 g/l (concentrate), and the metal ions were determined by AAS method, and
concentrate obtained from desalting electrodi- the biomass as a dry weight at 105°C.
alysis (diluate). Current density of 67.7 mA/cm 2 Calculations. The equations were taken from
and voltage of 12 V were applied. The circulation L�� et al. (1998).
flow rate was 2 l/min. When conductivity in the
diluate reached the value of 5 mS/cm, the experi- RESULTS AND DISCUSSION
ments were terminated.
Ultrafiltration. UF cartridge TFF 300 kD (Mil- A sequence of five separative steps for the lactic
lipore, USA) was used for the clarification of the acid recovery from the fermentation broth was
fermented broth. proposed and tested. The separation consisted of
Decolourisation, the removal of multivalent three pre-treating steps (ultrafiltration, decolouri-
metal ions. These operations were carried out in sation, the removal of multivalent metal ions),
glass columns filled with the above mentioned ma- and of two electrodialysis procedures (desalting
terials. The flow rate of the cell-free fermentation electrodialysis and water-splitting electrodialysis)
broth through the column was 1 bed volume per (Figure 1).
hour (decolourisation), resp. 2 bed volumes per
hour (removing of multivalent metal ions). Pretreatment of fermentation broth
Analytical methods. Lactate was determined
by HPLC (Laboratorní přístroje Praha, Czech The fermentation broth had to be pretreated
Republic); column – Ostion LG KS in H-cycle; prior to the electrodialysis experiment due to the
refractometer detector RIDK 101; mobile phase high demands of the electrodialysis membranes,
– H2SO4 (c = 0.005 mol/l), flow rate of the mobile especially bipolar ones, on the quality of the feed
phase 0.5 ml/min; column temperature 85°C. The solution used. The cells can be deposited on the
concentration of lactic acid and the concentration membrane surface and create clusters in the space
of NaOH were determined by titration with stand- between the membranes, the dyes from the fermen-
ard solutions of NaOH (c = 0.025 mol/l) or HCl tation broth can be adsorbed on the membranes.
(c = 1 mol/l), respectively, using phenolphthalein as Both phenomena significantly decrease the elec-
indicator. The colour intensity of the fermentation trodialysis efficiency (C����� et al. 1987; H������
broth was measured by spectrophotometer at the et al. 1993). Multivalent metal ions (Ca, Mg, Fe,
wavelength of 400 nm relative to water. Multivalent etc.) cause irreversible damage of the bipolar

Pretratment of Lactic acid

fermentation broth: recovery:
PRETREATED
FERMENTATION BROTH FERMENTATION BROTH

Ultrafiltration Desalting
electrodialysis
CELL FREE PERMEATE

Decolourisation CONCENTRATED SODIUM LACTATE

DECOLOURISED PERMEATE
Water-splitting
Removing of electrodialysis
multivalent metal ions

Figure 1. Scheme of lactic acid recovery from

PRETREATED LACTIC ACID
FERMENTATION BROTH fermentation broth

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Vol. 22, No. 3: 87–94 Czech J. Food Sci.

membranes, they require, consequently, less than Table 1. Comparison of resins used for removal of mul-
1 mg multivalent metal ions per litre. tivalent metal ions (flow rate was 2 bed volumes/h)

Ion exchanger Purolite S 940 Duolite C-433

Ultrafiltration
Initial bivalent ions
1.845 1.845
The broth after lactic acid fermentation contained concentration (mmol/l)
3.35 g/l of biomass, the cells having been removed
Final bivalent ions
from it by a spiral-wound module. The cell free concentration (mmol/l)
0.0025 0.05
permeate and concentrate containing 51.1 g/l of
biomass were obtained. Relatively considerable Treated bed volumes 42 24
fouling of membranes was found. To use a differ- Retained ion
ent type of the membrane cartridge would seem 99.8 97.2
efficiency (%)
advisable.
Convenient for process yes no

Decolourisation

The renewable granulated active charcoal Purolite The compositions of the initial fermentation
AC 20 was chosen and used for decolourisation. The broth and of that treated with resin are given in
results were reported earlier (H����� et al. 2001). Table 2. Depleted broth contained less than 1 mg/l
The flow of the fermentation broth through the of multivalent metals and the bipolar membrane
column influenced significantly the decolourisa- could be used without risk of irreversible mem-
tion degree. If the flow rate was as slow as 1 bed brane damage.
volume per hour only, the amount of the fermenta-
tion broth corresponding to the 10-fold bed volume Electrodialysis
was decolourised by 90%. At a higher flow rate,
the decolourisation capacity decreased. When the Two-stage electrodialysis method was used for
decolourisation was carried out in a batch mode, lactic acid recovery. Sodium lactate was removed
the decolourisation degree was equal to that men- from the pretreated fermentation broth and was
tioned above; more decolourising agent, however, concentrated by desalting electrodialysis. Further
was used up. it was converted to lactic acid by water-splitting
Regeneration was done successively by water electrodialysis using bipolar membranes. Lactic
flow, by boiling with 4% HCl, by water flow, by acid was recovered from the model solutions and
boiling with 4% HCl and by water flow. Regenerated from the pretreated fermentation broth. The ex-
charcoal showed practically the same decolourisa- periments with model solutions were focused on
tion capacity as the initial batch and its value after the determination of suitable conditions for the
15 decolourising cycles was lower by 5%. electrodialysis experiments and on the investiga-
tion of the time course under different conditions.
Removal of multivalent metal ions The results obtained with model solutions were
published earlier (H����� et al. 2001, 2004).
Two ion exchangers were tested. The chelating
resin Purolite AC 20 G showed better results than Desalting electrodialysis
the weak cation exchanger Duolite C-433 (Table 1).
Purolite AC 20 G was used for the removal of mul- For desalting electrodialysis the fermentation
tivalent metal ions from the fermentation broth broth was used with three different levels of lactate
designated for the electrodialysis experiments. concentration: 18, 36, and 87 g/l. The results of the

Table 2. Composition of initial and treated fermentation broth (mg/l)

Ca Mg Fe Mn Zn Na K
Initial fermentation broth 28.3 38.8 0.64 4.36 0.44 16 500 935
Treated fermentation broth 0.39 0.47 0.04 0.02 0.02 16 800 868

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150 Figure 2. The time course of two

Lactate concentration (g/l)

level desalting electrodialysis

Concentrate of the pretreated fermentation
100 broth
Addition of the fresh
fermentation broth
50 into diluate circuit

Diluate

0
0 50 100 150 200 250 300 350 400

Time (min)

Table 3. Desalting electrodialysis experiments

Concentration (g/l)
Experiment

time (min)
Operating

Current Energy
VD /VK initial final initial final efficiency consumption
0 0
diluate diluate concentrate concentrate (%) (kWh/kg)

DM 1 120 1.87 18.1 1.3 3.9 29.2 53 0.41

DM 2 165 1.87 17.3 1.3 28.7 45.4 55 0.54
DM 3 195 1.87 22.2 1.2 43.0 60.6 39 0.53
DM 4 195 7.50 36.3 2.0 2.8 108 66 0.32
DM 5 180 7.50 36.9 2.8 104 152 69 0.32
DM 6 215 7.50 36.6 0.9 4.8 113 64 0.34
DM 7 195 7.50 36.2 1.2 107 147 64 0.34
DM 8 215 4.38 88.3 3.7 96.4 185 82 0.21
DM 9 220 4.38 85.8 1.2 93.6 175 81 0.22

VD – initial volume of diluate, VK – initial volume of concentrate

0 0

desalting electrodialysis experiments are shown in Nevertheless, at higher lactate concentrations in

Table 3. The highest lactate concentration reached the fermentation broth a very good agreement with
was 185 g/l. The comparison of the lactate recovery the model solutions was found.
from the model solutions and from the fermenta- Figure 2 shows the time course of two-level elec-
tion broth showed that, in the case of fermentation trodialysis of fermentation broth. The initial lactate
broth with lower initial lactate concentrations, the concentration was 36.6 g/l while the final one of
transport rate decreased, the energy consumption 146 g/l was obtained in the concentrate stream.
increased, and the current efficiency decreased. About 1 g lactate per litre remained in the diluate

Table 4. Water-spli�ing electrodialysis

Concentration (g/l) Energy

Operating Current
Experiment consumption
time (min) initial lactate final lactic acid final NaOH efficiency (%)
(kWh/kg)
WM 1 390 145 121 43 66 1.16
WM 2 430 171 151 53 79 0.95

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Vol. 22, No. 3: 87–94 Czech J. Food Sci.

150 Figure 3. The typical time

Lactate course of water-splitting
Lactic acid electrodialysis
Concentration (g/l)

100

50 NaOH

0
0 50 100 150 200 250 300 350 400

Time (min)

at the end of each level. The concentration of lactate The results confirm that two stage electrodialysis
obtained during the two-level electrodialysis was is a suitable and efficient technique for the recovery
about 4-times higher than that in the fermentation of lactate ions from the pretreated fermentation
broth. The current efficiency was 64% and the en- broth and the subsequent conversion into lactic
ergy consumption was 0.34 kWh/kg. acid with respect to environmental aspects.
Due to a high lactate concentration and the lowest In the first electrodialysis step, the final lactate
concentration of multivalent metal ions, concen- concentration of up to 175 g/l was obtained while the
trates DM 7 and DM 9 were used for the water- final lactic acid concentration of 151 g/l was reached
splitting electrodialysis experiments. The other in the second step. The total energy required in
concentrates had too low lactate concentrations both electrodialysis processes representing the
or contained more than 1 mg of multivalent metal energy consumption for the lactate transfer and
ions per litre and could not be used, consequently, for its electroconversion to lactic acid was about
for the bipolar membrane treatment. 1.5 kWh/kg of lactic acid obtained.

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Received for publication February 2, 2004
Accepted a�er corrections April 7, 2004

Souhrn

H����� V., M������ K., R������� M. (2004): Nová metoda separace kyseliny mléčné z fermentačního média.
Czech J. Food Sci., 22: 87–94.

Pro separaci kyseliny mléčné z fermentačního média byla použita dvoustupňová elektrodialýza. V prvním stupni
byl laktát sodný separován a nakoncentrován pomocí odsolovací elektrodialýzy a následně ve druhém stupni
byl pomocí elektrodialýzy s bipolárními membránami konvertován na kyselinu mléčnou. V prvním stupni bylo
v 1 litru získáno až 175 g laktátu a z něj ve druhém stupni 151 g kyseliny mléčné. Celkově bylo třeba na zisk 1 kg

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Vol. 22, No. 3: 87–94 Czech J. Food Sci.

kyseliny mléčné vynaložit 1,5 kWh elektrické energie. Před elektrodialýzou muselo být fermentační médium
upravováno. Úprava spočívala v ultrafiltraci, odbarvování a odstranění vícemocných iontů kovů.

Klíčová slova: izolace kyseliny mléčné; elektrodialýza; předúprava fermentačního média

Corresponding author:

Ing. V��� H�����, Vysoká škola chemicko-technologická v Praze, Fakulta potravinářské a biochemické
technologie, Ústav kvasné chemie a bioinženýrství, Technická 5, 166 28 Praha 6, Česká republika
tel.: + 420 224 354 127, fax: + 420 224 355 051, e-mail: [email protected]

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