TEPZZZ84B_T

(19)

(11) EP 2 038 422 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.:

of the grant of the patent: C12P 7/56 (2006.01) C13B 10/08 (2011.01)
26.11.2014 Bulletin 2014/48
(86) International application number:
(21) Application number: 07765568.6 PCT/EP2007/056256

(22) Date of filing: 22.06.2007 (87) International publication number:

WO 2008/000699 (03.01.2008 Gazette 2008/01)

(54) LACTIC ACID PRODUCTION FROM CONCENTRATED RAW SUGAR BEET JUICE

MILCHSUREPRODUKTION AUS KONZENTRIERTEM ROHEM ZUCKERRBENSAFT

PRODUCTION DACIDE LACTIQUE A PARTIR DE JUS BRUT DE BETTERAVE A SUCRE
CONCENTRE

(84) Designated Contracting States: ZAKHAROV, I. P. ET AL: "Production of lactic acid

AT BE BG CH CY CZ DE DK EE ES FI FR GB GR from sugar beet and cases of inactivation of lactic
HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE acid fermentation" MIKROBIOLOGIYA, vol. 15,
SI SK TR no. 1, 1946, pages 57-66, XP008071354 cited in
the application
(30) Priority: 26.06.2006 EP 06115885 HOLLAUS F ET AL: "Experimental studies on
bacterial degradation of sugar in raw juice and in
(43) Date of publication of application: preliming juice." SUCRERIE BELGE, vol. 99, no.
25.03.2009 Bulletin 2009/13 5, 1980, pages 183-193, XP008071302 cited in the
application
(73) Proprietor: PURAC Biochem BV DATABASE CA [Online] CHEMICAL ABSTRACTS
4206 AC Gorinchem (NL) SERVICE, COLUMBUS, OHIO, US; 1918,
BONELLI, A. ET AL: "The industrial preparation
(72) Inventors: of lactic acid from sugar beets" XP002407653
VISSER, Diana retrieved from STN Database accession no. 1919:
NL-4201 EP Gorinchem (NL) 714 cited in the application & IND. CHIM. MIN.
VAN BREUGEL, Jan MET. , 5, 121-4, 1918,
NL-2485 WN Woudrichem (NL) MANZKE E ET AL: "Raw thick juice: manufacture,
DE BRUIJN, Johannes Martinus storage and utilization as feedstock in the
NL-4813 GK Breda (NL) biotechnological industry" ZUCKERINDUSTRIE,
ACAMPO, Paul vol. 117, no. 12, 1992, pages 984-990,
NL-3755 BA Eemnes (NL) XP008071289 cited in the application
GOEKSUNGUR Y ET AL: "Batch and continuous
(74) Representative: Beetz, Tom et al production of lactic acid from beet molasses by
De Vries & Metman Lactobacillus delbrueckii IFO 3202" JOURNAL
Overschiestraat 180 OF CHEMICAL TECHNOLOGY AND
1062 XK Amsterdam (NL) BIOTECHNOLOGY, vol. 69, no. 4, 1 August 1997
(1997-08-01), pages 399-404, XP000702340 ISSN:
(56) References cited: 0268-2575 cited in the application
WO-A-00/56912 WO-A2-2006/001034
EP 2 038 422 B1

US-A- 1 969 237 US-B1- 6 440 222

Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent
Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the
Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been
paid. (Art. 99(1) European Patent Convention).

Printed by Jouve, 75001 PARIS (FR) (Cont. next page)

 EP 2 038 422 B1

GOEKSUNGUR Y ET AL: "Production of lactic EL-SHERBINY G A ET AL: "Utilization of beet

acid from beet molasses by calcium alginate molasses in the production of lactic acid"
immobilized Lactobacillus delbrueckii IFO 3202" EGYPTIAN JOURNAL OF FOOD SCIENCE, vol.
JOURNAL OF CHEMICAL TECHNOLOGY AND 14, no. 1, 1986, pages 91-100, XP008071367 ISSN:
BIOTECHNOLOGY, vol. 74, no. 2, February 1999 0301-8571 cited in the application
(1999-02), pages 131-136, XP000830050 ISSN: KOTZAMANIDIS C ET AL: "Optimization of lactic
0268-2575 cited in the application acid production from beet molasses by
MONTEAGUDO J M ET AL: "Kinetics of lactic acid Lactobacillus delbrueckii NCIMB 8130." WORLD
fermentation by Lactobacillus delbrueckii grown JOURNAL OF MICROBIOLOGY &
on beet molasses" JOURNAL OF CHEMICAL BIOTECHNOLOGY, vol. 18, no. 5, 2002, pages
TECHNOLOGY AND BIOTECHNOLOGY, vol. 68, 441-448, XP008071357
no. 3, 1 March 1997 (1997-03-01), pages 271-276,
XP000700646 ISSN: 0268-2575 cited in the
application
MONTEAGUDO J M ET AL: "Optimization of the
conditions of the fermentation of beet molasses
to lactic acid by Lactobacillus delbrueckii" ACTA
BIOTECHNOLOGICA, vol. 14, no. 3, 1994, pages
251-260, XP002407024 ISSN: 0138-4988

2
 EP 2 038 422 B1

Description

[0001] The present invention is in the field of the preparation of lactic acid by means of fermentation.
[0002] Lactic acid, its salts and esters have long been used as food additive and in various chemical and pharmaceutical
5 applications. More recently, lactic acid has been used in the making of biodegradable polymers both as a replacement
for present plastic materials as well as various new uses where biodegradability is needed or desired such as for medical
implants, solvable sutures and controlled release drugs.
The production of lactic acid is commonly carried out by fermentation by means of a micro-organism such as bacteria,
yeasts and fungi. The fermentation medium consists of a carbohydrate substrate together with suitable mineral and
10 proteinaceous nutrients. A commonly used fermentation substrate is white sugar. Sugar is the most important contributor
to the manufacturing cost price of lactic acid. Major reductions in the manufacturing cost price of lactic acid can therefore
be accomplished if a less expensive carbohydrate source can be used than white sugar. To this end several research
groups tried to ferment cheaper byproducts and intermediates of a sugar production plant to lactic acid. However, while
the fermentation of these crude sugar sources to ethanol can be done readily, problems are encountered when trying
15 to use these substrates on industrial scale for the fermentation to lactic acid. These problems lie in the field of fermentability,
storage - stability, sensitivity to infections, the purification of the product of fermentation (i.e. the downstream processing)
etcetera.
Examples of prior art wherein the fermentation of sugar beet juice to ethanol is described are Raw thick juice: manufacture,
storage and utilisation as feedstock in biotechnological industry, G. Marke, P.V. Schmidt, R. Rieck, B. Senge, B. Steiner,
20 Zuckerindustrie (1992), 117 (12), 984-90 , The manufacture of alcohol from sugar beets, K. Antal, Zeitschrift fuer Spir-
itusindustrie (1911) 34, 239-40, 252-3 and Combined production of ethanol and white sugar, K. Austmeyer, H Roever,
H. Zuckerindustrie (1988) 113 (9), 765-72.
The literature on fermentation of sugar beet juice to lactic acid on industrial scale is not so abundant.
[0003] For instance, in The industrial preparation of lactic acid from sugar beets, A. Bonelli, G.Gulinelli, Ind. Chim.
25 Met. (1918), 5 121-4, the fermentation of raw sugar beet juice to lactic acid is described. This raw sugar beet juice only
has a concentration of about 16 wt% sugar. First of all, as indicated in the publication this carbohydrate source is very
sensitive to infections and often already infected to start with. This is confirmed in for instance, F. Hollaus et AI:
" Experimental studies on bacterial degradation in sugar of sugar in raw juice and preliming juice." Sucrerie belge, vol
99, No 5, (1980), p. 183. With sugar beet factories running in campaigns and only active 3-4 months a year, it is clear
30 that this provides storage problems. Secondly, with a concentration of only 16% the transport and storage costs will
render this process relatively expensive. These factors make raw sugar beet juice unfit as a substrate for fermentation
to lactic acid on an industrial scale.
[0004] In Production of lactic acid from sugar beet and cases of inactivation of lactic acid fermentation, I.P. Zakharov,
M.F. Federova, Mikrobiologiya (1946), 15(No. 1), 57-66, also the fermentation of raw sugar beet juice is described.
35 Zakharov reports that the heating or sterilization of the sugar beet juice has a detrimental effect on the fermentation.
The beet syrup, prepared by evaporation of beet juice to a sugar content of 51% was diluted and fermentation was tried:
virtually no fermentation took place both with and without chalk (probably calcium hydroxide is meant here) addition.
Zakharov concluded that a medium of such syrup proved unsuitable for the fermentation to lactic acid.
[0005] All in all, it must be concluded that the fermentation of these crude substrates such as raw sugar beet juice to
40 lactic acid on industrial scale proves to be much more complicated than a comparable fermentation to ethanol. The
same can be said of the purification (i.e. the downstream processing) of lactic acid prepared by fermentation compared
to the purification of fermentatively prepared ethanol.
[0006] We have found that concentrated raw beet juice having a Brix of at least 60 (i.e. amount of sugar in weight per
100 grams of liquid) is a suitable substrate for the fermentation on industrial scale to lactic acid and or lactate. Further,
45 it appears to be storage-stable and is not very sensitive to infections. Furthermore, fermentation to lactic acid is achievable
with the same yield, chemical purity, optical purity, clarity and taste as lactic acid obtained from fermenting sucrose, i.e.
white sugar.
[0007] When processing sugar beet, the beet is usually washed with water, cut and the resulting cossettes are extracted
with water, from which beet pulp is removed and the resulting raw juice is further processed to sugar by subsequent
50 juice purification [i.e. traditionally by lime addition to a pH of approximately 11.2 in the pre-liming, followed by surplus
lime addition in the main liming (the alkalization), addition of carbon dioxide in two subsequent stages (the carbonation),
wherein after the first carbonation the carbonation sludge/lime is removed] resulting in thin juice, said thin juice is
concentrated resulting in thick juice, said thick juice is subjected to one or more crystallization steps to form various
sugar grades, and molasses as by-product.
55 [0008] The concentrated raw beet juice used in the process according to the invention is prepared by subjecting the
raw juice (approximately 16% sugar) to a heating step at a temperature between 50 and 90 C and concentrating the
raw beet juice to a Brix of at least 60 (that is, without lime addition to a pH of approximately 11.2 and surplus lime addition
followed by carbon dioxide addition in two stages with removal of carbonation sludge/lime). Optionally a liquid-solid

3
 EP 2 038 422 B1

separation step is performed between the heating step and the concentration step and/or after the concentration step
to remove residual soil, small beet particles and protein. The concentration step is usually performed by evaporation at
a temperature between 50 and 120 C. It is also possible to combine the heating and concentration step. If the heating
and concentration steps are combined, the optional solid / liquid separation is conducted either prior to the heating/con-
5 centration or after. The concentrated raw beet juice can be added in diluted form to the fermentation. Usually the
concentrated raw beet juice is diluted to a concentration of about 16-30 Brix, more preferably 20-30 Brix.
[0009] The processing of sugar beet to white sugar and the preparation of concentrated raw beet juice from the raw
beet juice which is an intermediate of the beet processing process is schematically illustrated by figure 1.
[0010] It was found that molasses and thick juice prepared during sugar beet processing is not very suitable for the
10 fermentation to lactic acid on industrial scale. Without wishing to be bound to a theory we think that during the processing
of the raw sugar beet juice to thick juice and further to molasses, several impurities are either concentrated in the
molasses and/or thick juice or are introduced by reactions occurring during the alkalization, heat and carbon dioxide
treatments (e.g. the results of Maillard reactions). Said impurities interfere with fermentation reactions to lactic acid on
industrial scale. This is also shown in our experimental data. Sometimes the impurities can be removed by various pre-
15 treatments, but this requires additional, laborious and expensive purification steps.
[0011] Goeksungur y et all: "Batch and continuous production of lactic acid from beet molasses lactobacillus delbrueckii
IFO 3202" Journ. Chem. Techn. and Biotechn., vol 69, No.4, (1997) pp. 399-404, and Goeksungur y et all:"Production
of lactic acid from beet molasses by calcium alginate immobilized lactobacillus delbrueckii IFO 3202" Journ. Chem.
Techn. and Biotechn., vol 74, No.2, (1999) pp. 131-136, describe the production of lactic acid from pretreated beet
20 molasses with specific bacteria. The pretreatment comprises acidification with sulphuric acid, boiling, centrifusion, fil-
tration and clarification, including pasteurization. The experiments were conducted in 250 cm3 and 500 cm3 flasks and
it was indicated that even with a low initial sugar concentration (28.2 g /l) the sugar was not completely utilized. The
authors attribute that to the complex nature of molasses (i.e. impurities).
[0012] Monteagudo J.M. et AI: " Kinetics of lactic acid fermentation by lactobacillus delbruekii grown on beet molasses",
25 Journ. Chem. Techn. And Biotechn., vol 68, No 3, (1997), pp 271-276 describes the experiments of lactic acid fermentation
on beet molasses on lab scale (5 liter flasks). Here also the sugar was not completely utiised.
[0013] EI Sherbiny et al:" Utilisation of beet molasses in the production of lactic acid", Egyptian Journ. Of Food SC.vol
14, No 1 (1986), pp 91-100, also shows that in the fermentation not all sugar is utilized and that several other organic
acids are formed besides lactic acid.
30 [0014] The industrial fermentation to lactic acid requires strict control of temperature and pH. Because of the formation
of lactic acid the pH drops during fermentation. A drop in pH below a critical value, depending on the microorganism
used in the process, could damage the microorganisms metabolic process and bring the fermentation process to a stop.
Therefore, it is common practice to add a neutralizing agent, i.e. a base such as Ca(OH)2 , Mg(OH)2, NaOH, KOH or
ammonia to the fermentation reaction and thus produce a lactate salt such as calcium lactate, sodium lactate etcetera.
35 Normally both lactic acid and lactate salt are present in the fermentation product, depending on the pH of the fermentation
product. The fermentation can be done with conventional lactic acid-producing microorganisms such as bacteria yeasts
and fungi, such as lactobacilli, moderately thermophilic bacilli, Rhizopus and Aspergillus. Preferred are the moderately
thermophilic bacilli such as Bacillus coagulans, Bacillus thermoamylovorans, Geobacillus stearothermophylus and Ba-
cillus smithii, because these types of micro-organisms can ferment at relatively high temperature.
40 [0015] After fermentation, the lactate and lactic acid-containing fermentation product must be separated from the
biomass. Said lactic acid-containing fermentation product is in the liquid form (i.e. liquid or in solution). Usually this
biomass is separated by means of filtration, centrifuging, flocculation, coagulation, flotation or combinations thereof.
After biomass separation, the pH of the fermentation product is decreased by means of acid addition such as sulphuric
acid so that lactic acid and a salt of the neutralizing base and the added acid is formed. For example, if calcium hydroxide
45 is used as neutralizing agent, the fermentation product will comprise both lactic acid and calcium lactate. Upon addition
of sulphuric acid, lactic acid and calcium sulphate (gypsum) will be formed. The gypsum, or any other salt, is removed
and the lactic acid is isolated. The resulting lactic acid can be subjected to further purification steps.
[0016] Conventional subsequent purification steps are distillation including short path distillation and vacuum distilla-
tion, crystallization, salt SWAP, electrodialysis, extraction (both forward and back-extraction), carbon treatment, ion
50 exchange and combinations thereof. These purification steps can be combined with intermediate concentration steps.
[0017] The invention is further illustrated by means of examples which are not to be interpreted as limitative.

Example 1

55 Fermentation to lactic acid using various substrates

[0018] The fermentability of molasses, thick juice and concentrated raw juice substrates originating from a sugar
factory (5 samples with Brix values varying from 59.6 to 73.2, concentrated at temperatures of 55, 60, 75, and 85 C),

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 EP 2 038 422 B1

was tested in separate fermentations. For all substrates, the following procedure was applied: the substrate was diluted
with water to a sucrose concentration of approximately 300 g/l. To this solution, 6.0 g/l nutrients were added and the pH
was adjusted to 6.4 using lime. Subsequently, the medium was heated to 54C and the fermentation was started with
10% (v/v) of a sucrose-grown inoculum of a lactic acid-producing microorganism.
5 [0019] During the fermentation diluted lime was added to keep pH at 6.4 and the temperature was controlled at 54C.
A fermentation in which white sugar (sucrose) was used as substrate, and otherwise applying the same conditions, was
carried out for comparison.
[0020] Table 1 summarizes the results of these fermentations. Fermentations based on molasses hardly showed any
activity; only a small fraction of the sugar was consumed after 45 hrs and the productivity collapsed to virtually zero. The
10 fermentability of thick juice was more active, but still only a part of the sugar was consumed. Concentrated raw juice
could be fermented completely. Compared to the reference sucrose fermentation, the lactic acid yield and byproduct
profile (organic acids) is similar; the chiral purity is lower but still acceptable. The residual sugar content is higher, because
concentrated raw juice contains some sugars that are not/slowly fermented.

15 Table 1. Summary fermentation results for sucrose (reference), molasses, thick juice and concentrated raw juice.
Substrate Composition final broth
Lactate (% w/w of Chiral purity Residual sugars (% Polysaccharides Total organic
max. obtainable) lactate (%S) w/w of sugar input) (% w/w of lactate acids* (%w/w)
20 formed)
Sucrose*** 95% 99.7% 1% 0.9% n.d.
Molasses n.d. n.d. 100% n.d. n.d.
Thick juice n.d. n.d. >50% n.d. n.d.
25
Concentrated 95% 99.2% 3.5% 2.3% <0.2%
raw juice**
* Sum of succinic, pyruvic, formic, 2-hydroxybutyric and acetic acid
** Average of 11 fermentations
30 *** Average of 13 fermentations
n.d. = not determined

Example 2
35
Concentrated raw sugar beet juice in fermentation on industrial scale

[0021] Two semi-industrial fermentations (scale 5000 liters) on concentrated raw beet juice were performed. Of the
final fermentation broth of these two fermentations biomass was removed and the lactate-containing product was acid-
40
ulated.

A. Materials and Methods

Substrate
45
[0022] Sugar beets were cut, washed and extracted in water at 70C. The pulp was eliminated and the raw beet
juice was subjected to heat treatment at 60 C, solid / liquid separation and evaporation at 65C to make concentrated
raw beet juice of a Brix between 65-70.
The density of the batches of concentrated raw beet juice was 1.334 kg/l and 1.335 kg/l at 66 Brix, respectively.
50
Fermentation

[0023] Fermentation of concentrated raw beet juice was carried out in the 5000 l fermenter using a lactic acid-producing
microorganism. The medium composition of the fermentation is shown in table 2.
55

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 EP 2 038 422 B1

Table 2. The 5 m 3 reactor received a medium of the following composition:

Medium composition added amount
Water 2000 l
5
Concentrated raw beet juice (appr. 70Bx) 1000 l
Nutrient 18.16 kg
Start volume 3000 l

10 Inoculum 500 l

[0024] The temperature during the fermentation was kept at 54C. The pH was controlled by adding a slurry of lime.
[0025] Both fermentations started with 275 g/l sucrose (and a total amount of approximately 3 g/l glucose and fructose).

15 Biomass removal

[0026] Flocculation, to remove bacterial cells, metals, proteins etc., was performed with sugar free final fermentation
broth by alkalizing the broth at a temperature of 75 C with lime and adding flocculant.
The clear top layer was siphoned off as soon as the settling of the biomass was complete. The clear alkaline top layer
20 was subsequently transferred to a second 5 m 3 reactor for acidulation. The biomass containing bottom layer was dis-
carded.

Acidulation

25 [0027] The clear alkaline top layer was continuously stirred and sulphuric acid (96%) was added slowly using a pump.
This was continued until a pH of 2.2 and a conductivity of approximately 5 mS was reached. Precipitation of calcium
was checked using an ammonium oxalate solution (35.5 g/l). The acidulated broth was removed from the reactor and
isolated from the formed gypsum.
The clear crude lactic acid solution was subjected to conventional purification steps to make final S-lactic acid (90%).
30

Results

[0028] Both fermentations performed as expected. The culture followed the growth phase and the stationary or pro-
duction phase until sugar free. The first fermentation took 24 hours. The second fermentation finished after 21 hours.
35 Final fermentation broth of the first fermentation had a substantial higher amount of polysaccharides than final fermen-
tation broth of the second fermentation. The difference in the amount of total nitrogen in final fermentation broth from
these two batches, is negligible. Optical purity of final fermentation broth was 99.3 % for the first fermentation and 99.4
% for the second fermentation.
The results of the analyses of final fermentation broth samples are listed in table 3. The results show that concentrated
40 raw sugar beet juice can be fermented to lactic acid on industrial scale.

Table 3. Lactate, total residual sugars, polysaccharides and optical purity of final fermentation broth
5m3 final fermentation Lactate (% w/w Total residual sugars Polysaccharides (% w/w of Chiral purity
broth of max. (% w/w of input) lactate) (%S-lactate)
45
obtainable)
1st 95% 4.2% 4.0% 99.3%
2nd 95% 6.5% 2.6% 99.4%

50
[0029] After purification, the resulting lactic acid appeared to fulfill the specifications set for commercial lactic acid for
food applications with respect to color, taste and optical purity.

55 Claims

1. Process for the preparation of lactic acid and/or lactate wherein a concentrated raw beet juice, i.e. beet juice which
has not been submitted to lime addition to a pH of approximately 11.2 and surplus lime addition followed by carbon

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 EP 2 038 422 B1

dioxide addition in two stages with removal of carbonation sludge/lime, having a concentration of at least 60 g
sucrose per 100 gram liquid (at least a Brix of 60) is used as a substrate for the fermentation to lactic acid and/or
lactate by means of a lactic acid-producing micro-organism.

5 2. Process according to claim 1 wherein the concentrated raw beet juice has a Brix of at least 70.

3. Process according to claim 1 or 2 wherein the concentrated raw beet juice is diluted to a Brix between 16 and 30
prior to addition to the fermentation.

10 4. Process according to any one of the preceding claims wherein the microorganism is a moderately thermophilic
Bacillus.

5. Process according to any one of the preceding claims wherein the concentrated raw beet juice is prepared by a
procedure comprising heat treating a raw beet juice at a temperature between 50 and 90 degrees Celsius and
15 concentrating the raw beet juice to at least 60 Brix.

6. Process according to claim 5 wherein the concentrated raw beet juice is prepared by a procedure comprising the
following steps:

20 a. washing and cutting sugar beet to obtain cossettes and extracting the cossettes with water,
b. removing beet pulp from the resulting raw beet juice, and
c. heat treating the raw beet juice at a temperature between 50 and 90 degrees Celsius, and
d. concentrating the raw beet juice to at least 60 Brix.

25 7. Process according to claims 5 or 6 wherein the heat treatment step and the concentration step are combined.

8. Process according to claims 5, 6 or 7 wherein prior to the heat treatment step and/or between the heat treatment
step and the concentration step, and/or after the concentration step a liquid / solid separation step is conducted.

30 9. Process according to any one of the preceding claims wherein the lactic acid and/or lactate resulting from the
fermentation is subjected to one or more purification steps.

10. Process according to claim 9 wherein the purification steps comprise biomass separation, extraction, salt SWAP,
distillation, ion exchange, carbon treatment, extraction, and/or concentration and combinations thereof.
35

Patentansprche

1. Verfahren zur Herstellung von Milchsure und/oder Lactat, wobei ein konzentrierter roher Rbensaft, d.h. Rbensaft,
40 der keiner Zugabe von Kalk auf einen pH von etwa 11,2 und Zugabe von berschssigem Kalk, gefolgt von der
Zugabe von Kohlendioxid in zwei Stufen unter Entfernung von Carbonisierungsschlamm/Kalk, unterzogen wurde,
mit einer Konzentration von mindestens 60 g Saccharose pro 100 Gramm Flssigkeit (einem Brix von mindestens
60) als Substrat zur Fermentierung zu Milchsure und/oder Lactat mit Hilfe eines Milchsure produzierenden Mi-
kroorganismus verwendet wird.
45
2. Verfahren nach Anspruch 1, wobei der konzentrierte rohe Rbensaft einen Brix von mindestens 70 aufweist.

3. Verfahren nach Anspruch 1 oder 2, wobei der konzentrierte rohe Rbensaft vor der Zugabe zur Fermentierung auf
einen Brix zwischen 16 und 30 verdnnt wird.
50
4. Verfahren nach einem der vorhergehenden Ansprche, wobei der Mikroorganismus ein mig thermophiler Bacillus
ist.

5. Verfahren nach einem der vorhergehenden Ansprche, wobei der konzentrierte rohe Rbensaft durch ein Verfahren
55 hergestellt wird, das Wrmebehandlung eines rohen Rbensaftes bei einer Temperatur zwischen 50 und 90C und
Konzentrieren des rohen Rbensaftes auf mindestens 60 Brix umfasst.

6. Verfahren nach Anspruch 5, wobei der konzentrierte rohe Rbensaft durch ein Verfahren hergestellt wird, das die

7
 EP 2 038 422 B1

folgenden Schritte umfasst:

a. Waschen und Schneiden von Zuckerrbe, um Schnitzel zu erhalten, und Extrahieren der Schnitzel mit Wasser,
b. Entfernen von Rbenpulpe aus dem resultierenden rohen Rbensaft, und
5 c. Wrmebehandeln des rohen Rbensaftes bei einer Temperatur zwischen 50 und 90C, und
d. Konzentrieren des rohen Rbensaftes auf mindestens 60 Brix.

7. Verfahren nach den Ansprchen 5 oder 6, wobei der Wrmebehandlungsschritt und der Konzentrationsschritt kom-
biniert sind.
10
8. Verfahren nach den Ansprchen 5, 6 oder 7, wobei vor dem Wrmebehandlungsschritt und/oder zwischen dem
Wrmebehandlungsschritt und dem Konzentrationsschritt und/oder nach dem Konzentrationsschritt ein Flssig-
Fest-Trennschritt durchgefhrt wird.

15 9. Verfahren nach einem der vorhergehenden Ansprche, wobei die Milchsure und/oder das Lactat, die aus der
Fermentation resultieren, einem oder mehreren Reinigungsschritten unterzogen werden.

10. Verfahren nach Anspruch 9, wobei die Reinigungsschritte Biomassentrennung, Extraktion, Salzaustausch, Destil-
lation, Ionenaustausch, Behandlung mit Kohlenstoff, Extraktion und/oder Konzentration und Kombinationen davon
20 umfassen.

Revendications

25 1. Procd de prparation dacide lactique et/ou de lactate, caractris en ce quun jus de betterave brut concentr,
cest--dire un jus de betterave qui na pas t soumis laddition de citron vert un pH denviron 11,2 et laddition
de citron vert en surplus suivie par laddition de dioxyde de carbone en deux tapes avec llimination du citron
vert/boue de carbonatation, ayant une concentration dau moins 60 g de saccharose pour 100 g de liquide (au moins
60 Brix) est utilis comme substrat pour la fermentation en acide lactique et/ou lactate au moyen dun micro-
30 organisme de production dacide lactique.

2. Procd selon la revendication 1, caractris en ce que le jus de betterave brut concentr a un Brix dau moins 70.

3. Procd selon la revendication 1 ou 2, caractris en ce que le jus de betterave brut concentr est dilu un Brix
35 entre 16 et 30 Brix avant laddition pour la fermentation.

4. Procd selon lune quelconque des revendications prcdentes, caractris en ce que le micro-organisme est
un bacille modrment thermophile.

40 5. Procd selon lune quelconque des revendications prcdentes, caractris en ce que le jus de betterave brut
concentr est prpar par une procdure comprenant le traitement thermique dun jus de betterave brut une
temprature entre 50 et 90 C et la concentration du jus de betterave brut au moins 60 Brix.

6. Procd selon la revendication 5, caractris en ce que le jus de betterave brut concentr est prpar par une
45 procdure comprenant les tapes suivantes :

a. laver et couper la betterave sucre pour obtenir des cossettes et extraire les cossettes avec de leau,
b. retirer la pulpe de betterave du jus de betterave brut rsultant, et
c. traiter thermiquement le jus de betterave brut une temprature entre 50 et 90 C, et
50 d. concentrer le jus de betterave brut au moins 60 Brix.

7. Procd selon la revendication 5 ou 6, caractris en ce que ltape de traitement thermique et ltape de con-
centration sont combines.

55 8. Procd selon la revendication 5, 6 ou 7, caractris en ce que, avant ltape de traitement thermique et/ou entre
ltape de traitement thermique et ltape de concentration, et/ou aprs ltape de concentration, une tape de
sparation liquide/solide est ralise.

8
 EP 2 038 422 B1

9. Procd selon lune quelconque des revendications prcdentes, caractris en ce que lacide lactique et/ou le
lactate rsultant de la fermentation est soumis une ou plusieurs tapes de purification.

10. Procd selon la revendication 9, caractris en ce que les tapes de purification comprennent la sparation de
5 la biomasse, lextraction, lchange de sel, la distillation, lchange dions, le traitement carbonique, lextraction et/ou
la concentration et des combinaisons de ces tapes.

10

15

20

25

30

35

40

45

50

55

9
EP 2 038 422 B1

10
 EP 2 038 422 B1
REFERENCES CITED IN THE DESCRIPTION
This list of references cited by the applicant is for the readers convenience only. It does not form part of the European
patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be
excluded and the EPO disclaims all liability in this regard.
Non-patent literature cited in the description
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K. ANTAL. The manufacture of alcohol from sugar
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239-40, 252-3 [0002]
K. AUSTMEYER ; H ROEVER. Combined produc-
tion of ethanol and white sugar. H. Zuckerindustrie,
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A. BONELLI ; G.GULINELLI. The industrial prepa-
ration of lactic acid from sugar beets. Ind. Chim. Met.,
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F. HOLLAUS. Experimental studies on bacterial deg-
radation in sugar of sugar in raw juice and preliming
juice. Sucrerie belge, 1980, vol. 99 (5), 183 [0003]
11
I.P. ZAKHAROV ; M.F. FEDEROVA. Production of
lactic acid from sugar beet and cases of inactivation
of lactic acid fermentation. Mikrobiologiya, 1946, vol.
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GOEKSUNGUR Y. Batch and continuous production
of lactic acid from beet molasses lactobacillus del-
brueckii IFO 3202. Journ. Chem. Techn. and Bio-
techn., 1997, vol. 69 (4), 399-404 [0011]
GOEKSUNGUR Y. Production of lactic acid from
beet molasses by calcium alginate immobilized lacto-
bacillus delbrueckii IFO 3202. Journ. Chem. Techn.
and Biotechn., 1999, vol. 74 (2), 131-136 [0011]
MONTEAGUDO J.M. Kinetics of lactic acid fermen-
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EI SHERBINY et al. Utilisation of beet molasses in
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