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discussions, stats, and author profiles for this publication at: http://www.researchgate.net/publication/227927610

Unstructured kinetic model for reuterin and

1,3‐propanediol production by Lactobacillus
reuteri from glycerol/glucose cofermentation

ARTICLE in JOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY · MAY 2009

Impact Factor: 2.49 · DOI: 10.1002/jctb.2098

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Research Article
Received: 11 September 2008 Revised: 28 October 2008 Accepted: 29 October 2008 Published online in Wiley Interscience: 24 December 2008

(www.interscience.wiley.com) DOI 10.1002/jctb.2098

Unstructured kinetic model for reuterin and

1,3-propanediol production by Lactobacillus
reuteri from glycerol/glucose cofermentation
Montserrat Tobajas,∗ Angel F. Mohedano, José Antonio Casas and
Juan José Rodrı́guez

Abstract
BACKGROUND: Lactobacillus reuteri is unable to grow on glycerol as sole carbon and energy source hence, glycerol is used
as an alternative hydrogen acceptor during growth on available carbohydrates. Thus, glycerol is converted to reuterin and
1,3-propanediol (1,3-PDL), both products with interesting industrial applications. These compounds are commonly produced
by using resting cells in two-step fermentation processes.

RESULTS: The glycerol/glucose cofermentation by L. reuteri yields reuterin and 1,3-PDL at a glycerol concentration higher than
100 mmol L−1 . An increase of glycerol concentration from 200 to 400 mmol L−1 showed no additional stimulatory effect on
ethanol and acetate production but consistently reduced the lactate concentration. It was also found that reuterin concentration
reached a maximum value and subsequently decreased due to its conversion to 1,3-PDL. An unstructured kinetic model was
proposed to describe simultaneously microbial growth, substrates consumption and products formation. A multi-response
nonlinear regression analysis based on Marquardt algorithm combined with a Runge-Kutta integration method was used to
obtain the values of the fitting parameters.

CONCLUSIONS: The optimum concentration of glycerol for maximum reuterin and 1,3-PDL production was 200 mmol L−1 . The
complete process was satisfactorily described by the kinetic model proposed.

c 2008 Society of Chemical Industry

Keywords: kinetic model; Lactobacillus reuteri; reuterin; 1,3-propanediol; cofermentation

NOTATION YXS yield of biomass on glucose (g mmol−1 )

−1
A acetate concentration (mmol L )
E ethanol concentration (mmol L−1 )
G glycerol concentration (mmol L−1 )
kX kinetic constant for growth rate in Equation (1) (L
mmol−1 h−1 )
k1 kinetic constant for growth rate in the glycerol/glucose
cofermentation (h−1 )
k2 saturation constant in Equation (6) (mmol L−1 )
k3 kinetic constant for reuterin production (L g−1 h−1 )
k4 kinetic constant for 1,3-propanediol production from
glycerol (L g−1 h−1 )
k5 kinetic constant for 1,3-propanediol production from
reuterin (L g−1 h−1 )
L lactate concentration (mmol L−1 )
P 1,3-propanediol concentration (mmol L−1 )
R reuterin concentration (mmol L−1 )
S glucose concentration (mmol L−1 )
t time (h)
X biomass concentration (g L−1 )
YAX yield of acetate on biomass (mmol g−1 )
YEX yield of ethanol on biomass (mmol g−1 )
YXG yield of biomass on glycerol (g mmol−1 )
INTRODUCTION
Lactobacillus reuteri, a heterofermentative lactic acid bacterium
naturally occurring in the intestinal flora of humans and other
species of animals, transforms glucose to lactate, acetate, ethanol
and carbon dioxide. Furthermore, L. reuteri can use glycerol as an
external hydrogen acceptor source in anaerobic fermentation
of glucose. Thus, glycerol is converted to reuterin and 1,3-
propanediol (1,3-PDL), both products with interesting industrial
applications. Reuterin, which occurs in three different forms (as
a monomer, as a hydrated monomer and as a dimer form of
β –hydroxypropionaldehyde),1 inhibits the growth of potentially
pathogenic microorganisms such as both gram-positive and gram-
negative bacteria, viruses, yeast and fungi.2 Hence, reuterin can
be used in the sterilization and fixation of biological tissues in the
manufacture of biocompatible implants in medicine3 and in the
∗ Correspondence to: Montserrat Tobajas, Sección de Ingenieria Quimica,
FacultaddeCiencias,UniversidadAutonomadeMadrid,CarreteradeColmenar,
km.15, 28049 Madrid, Spain. E-mail: montserrat.tobajas@uam.es
Sección de Ingenieria Quimica, Facultad de Ciencias, Universidad Autonoma
675

YLX yield of lactate on biomass (mmol g−1 ) de Madrid, Carretera de Colmenar, km.15, 28049 Madrid, Spain

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c 2008 Society of Chemical Industry
 www.soci.org
food industry as a biopreservative.4,5 Reuterin is also a precursor in
the production of industrial chemicals such as plastics and acrylic
acid.6 1,3-PDL can be used as a monomer for polycondensations
to produce polyethers, polyurethanes and polyesters, especially
in the manufacture of poly(trimethylene terephthalate) which
exhibits desirable properties in carpets and textiles.7
Reuterin is commonly produced by using resting cells in two-step
fermentation processes. First, L. reuteri cells are grown anaer-
obically on glucose and reuterin is subsequently produced by
washed cells resuspended and incubated in glycerol-containing
media. Currently 1,3-PDL is obtained using two different petro-
chemical processes. Shell developed a chemical route for 1,3-PDL
through the reaction of ethylene oxide with carbon monoxide and
hydrogen while Degussa uses the conventional route based on
the hydrolysis of acrolein followed by catalytic hydrogenation.8
Over recent years, extensive work has been done to increase
the productivity and yield of biotechnological processes able to
generate 1,3-PDL.9 The biological system developed by DuPont
based on transformed E. coli leads to 1,3-PDL using sugar from
corn and other crops, with a yield of 34% after 74 h.10 1,3-PDL can
also be obtained by the fermentation of glycerol with higher yield.
Depending on the price of glycerol, the latter process could still
be favoured over the former. Nowadays, the use of agricultural
areas for producing non-food materials, such as biodiesel, has led
to a simultaneous surplus of glycerol which may reduce its market
price and, hence, improve the economic feasibility of microbial
glycerol-to-propanediol conversion.11
It has been found that growing cells of L. reuteri can also
produce reuterin and 1,3-PDL by glycerol fermentation while
metabolizing carbohydrates, in particular glucose.12 In fact,
glycerol/glucose cofermentation leads to higher production of
1,3-PDL than that achieved by resting cells in glycerol as sole
carbon source.13 The presence of glycerol and its transformation
into 1,3-PDL enables the cells to recover the NAD+ used
during glycolysis. Furthermore, other products with potential
economical applications are also obtained since the bioconversion
of glycerol to reuterin and 1,3-PDL was found to influence the
metabolism of the sugar cofermented by altering the end product
distribution.14,15
Although the production of reuterin and 1,3-PDL by Lactobacilli
has been studied by several authors, the development of kinetic
models for L. reuteri growth, reuterin and 1,3-PDL production has
only been treated previously in resting cells.13 Kinetic models
describing cell growth and products such as lactic acid and
bacteriocins formation rates have been reported for L. reuteri,16
and other Lactobacillus strains17 – 21 as well as for Lactococcus,
Streptococcus and Pedioccocus strains.7,22 Rasch et al.16 developed
a deterministic model which was able to predict the oscillatory
behaviour during reuterin production by L. reuteri in a chemostat
and to describe satisfactorily the evolution of glucose and glycerol
concentrations but not those of biomass and reuterin in a batch
system. Monteagudo et al.17 studied the kinetics of conversion
of beet molasses to lactic acid by L. delbrueckii CECT 286 at
controlled pH and temperature. As they stated, the proposed
model simplified the true process. The biokinetics of cell growth
and bacteriocin production by L. amylovorus DCE 471 in SSM was
described as a function of the temperature and pH.21 Vázquez
and Murado7 proposed a simple model to describe the kinetics
of biomass growth, bacteriocins and lactic acid production by
Lactococcus lactis and Pediococcus acidilactici in a batch system.
However, the development of kinetic models describing microbial
676
growth, substrates consumption and products formation during
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c 2008 Society of Chemical Industry
M. Tobajas et al.
glycerol/glucose cofermentation by L. reuteri or other Lactobacilli
has not been found in the literature.
The aim of this work is to optimize reuterin and 1,3-PDL
production from the cofermentation of glycerol and glucose by
L. reuteri PRO 137. A kinetic model describing simultaneously cell
growth, substrates consumption and products formation during
glycerol/glucose cofermentation by L. reuteri is proposed.
MATERIALS AND METHODS
Microbiological methods
Lactobacillus reuteri PRO 137, from the Instituto Nacional de
Investigación y Tecnologı́a Agraria y Alimentaria (INIA, Spain),
was used in this study. Stock cultures were maintained at −40 ◦ C
in MRS medium23 supplemented with 15% (v/v) glycerol. L. reuteri
was transferred to a 250 mL Erlenmeyer flask containing 100 mL of
MRS broth and propagated at 37 ◦ C and 200 rpm under anaerobic
conditions in a thermostatized orbital shaker (Julabo mod. SW2L,
Seelbach, Germany).
Batch cultures were inoculated with 2% (v/v) cells from the late
growth phase. The culture medium employed was MRS broth
containing glucose (20 g L−1 ) as carbon source and various
concentrations of glycerol, ranging from 100 to 400 mmol L−1 .
MRS medium also contains 4 g yeast extract, 10 g bactopeptone,
8 g meat extract, 2 g K2 HPO4 , 2 g di-ammonium hydrogen citrate,
5 g sodium acetate, 200 mg MgSO4 .7H2 O, 50 mg MnSO4 .4H2 O
and 1 g Tween 80 L−1 .
Batch cofermentation runs of glycerol/glucose were carried
out in a 2 L stirred tank bioreactor (BIOSTAT B2, B. Braun Biotech
International, Melsungen, Germany) with pH, temperature, stirring
rate and dissolved oxygen concentration control. The operation
conditions were 40 ◦ C, 250 rpm and pH 5.5, which were found
to be optimum for growth of L. reuteri PRO 137 in a previous
work.13 The pH was maintained at 5.5 by automatic addition of
3 mol H3 PO4 L−1 or 3 mol NaOH L−1 . Anaerobic conditions were
established by continuously flushing with filter-sterilized nitrogen.
Cultures were carried out for 12 h and samples were taken at
regular intervals to evaluate biomass, substrates and products
concentrations. The cells were removed by centrifugation (Orto
Alresa, mod. Digicen, Madrid, Spain) at 4300 g for 10 min and the
supernatants were filtered through 0.22 µm pore size filters and
stored at −40 ◦ C for subsequent analyses.
Analytical methods
Biomass concentration was determined by optical density mea-
surements at 600 nm using a spectro-photometer (Varian, mod.
Cary 50 conc., California, USA). It was converted to cell dry weight
(CDW) using a previously obtained calibration curve. The pres-
ence of reuterin in the cell-free supernatant was detected by the
colorimetric method of Circle et al.24 as modified by Lüthi-Peng
et al.25 Absorbance reading (A560 nm ) was determined and reuterin
was quantified by comparing the values obtained with those of
an acrolein standard curve.26 Glucose, glycerol, ethanol, acetate,
lactate and 1,3-PDL were analyzed by high performance liquid
chromatography (HPLC) (Varian, California, USA). Separation pro-
ceeded at 65 ◦ C on a Metacarb 67 H column (300 mm × 6.5 mm)
using 0.025 N H2 SO4 as the mobile phase at a flow-rate of 0.6 mL
min−1 . Components were identified and quantified by refractive
index measurements with suitable standards.
J Chem Technol Biotechnol 2009; 84: 675–680
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RESULTS AND DISCUSSION 360 2.5

Metabolite distribution
300

Concentration (mmol L-1)

L. reuteri produces lactate, ethanol, acetate and CO2 from glucose. 2.0
glucose
The evolution of substrate and products concentrations during lactate

Biomass (g L-1)
acetate
the growth of L. reuteri on glucose is shown in Fig. 1. Acetate was 240 ethanol
biomass 1.5
initially introduced with the MRS medium at 65 mmol L−1 . The glycerol
180 reuterin
increase of biomass was coupled to the metabolism of glucose 1,3-PDL
and the production of organic acids and ethanol. The major end 1.0
product was lactate (108 mmol L−1 ) while acetate was produced at 120
a low level. Neither 1,3-PDL nor reuterin were detected when using 0.5
60
glucose as sole carbon source. A similar fermentation pattern was
reported by de Gines et al.27 for other L. reuteri strains (CRL 1098,
0 0.0
CRL 1100, and CRL 1101). 0 2 4 6 8 10 12
The initial glycerol concentration range (100–400 mmol L−1 ) was Time (h)
established according to previous studies found in the literature
indicating that reuterin production is not started until a certain Figure 2. Time-evolution of biomass, substrates and products along the
cofermentation of 111 mmol L−1 glucose and 350 mmol L−1 glycerol by L.
level of glycerol, i.e. 50 mmol L−1 , is exceeded,16 as we have
reuteri PRO 137.
confirmed (data not shown), and that the molar ratio of glucose to
glycerol should be no greater that 0.33 to favour the accumulation
of reuterin with lactobacilli.6 The effect of glycerol addition on of glycerol. These results are in agreement with those found by
the end products profile can be seen in Fig. 2, which shows the Oude et al.28 in the cofermentation of lactic acid and glucose
results obtained from the cofermentation of 111 mmol L−1 glucose by L. Buchneri and L. parabuchneri where the microbial growth
and 350 mmol L−1 glycerol by L. reuteri PRO137. Looking at the was essentially due to glucose degradation and only slightly to
evolution of biomass, no significant differences are observed with the degradation of lactic acid. However, glycerol consumption
respect to the experiments performed using only glucose (Fig. 1). resulted in the production of 1,3-PDL and reuterin although the
This suggests that biomass growth mainly occurred at the expense residual concentration of glycerol remained high after 12 h. As can
of glucose and possibly small amounts of other carbon compounds be seen in Fig. 2, reuterin concentration reached a maximum value
initially present in the medium and was not due to the degradation at 6 h and decreased at longer fermentation times. In addition to
this, the presence of glycerol altered the end products profile
by reducing ethanol and lactate formation by 61.7% and 22.3%,
120 2.5
respectively, and increasing the acetate production from 20 to
107 mmol L−1 compared with those of cells grown on glucose
100
Concentration (mmol L-1)

2.0 alone.
Table 1 shows biomass, substrates and products concentrations
Biomass (g L-1)

80
1.5 reached during glycerol/glucose cofermentation within the range
60 of initial glycerol concentration investigated. Increasing the
glucose glycerol concentration from 200 to 400 mmol L−1 had no
lactate
1.0
40 acetate
significant stimulatory effect on ethanol and acetate production
ethanol but monotonically reduced lactate concentration. Increasing the
biomass 0.5 initial glycerol/glucose molar ratio from 0.9 to 3.6 resulted in a
20
reduction of lactic acid production by 39%. The lactate/acetate
0 0.0 ratio was about 5.5 when growing L. reuteri on glucose alone
0 2 4 6 8 10 12 and varied from 1.7 to 0.7 during cofermentation of glucose with
Time (h) 100–400 mmol L−1 glycerol. The ratio of organic acids to ethanol
Figure 1. Time-evolution of biomass, glucose and products in a batch was 1.25 during the growth of L. reuteri on glucose whereas a value
culture of L. reuteri PRO 137 in MRS. Experimental (symbols) and fitted around 4.5 was obtained in the cofermentation of glucose and
(solid line) results. glycerol regardless of the initial glycerol concentration, between

Table 1. Biomass, glycerol and products concentration during glucose/glycerol cofermentation (S0 = 111 mmol L−1 )
Residual
Glycerol Biomass Ethanol Acetate Lactate glycerol Reuterin 1,3-PDL
(mmol L−1 ) (g L−1 ) (mmol L−1 ) (mmol L−1 ) (mmol L−1 ) (mmol L−1 ) (mmol L−1 ) (mmol L−1 )

0 2.2 102.0 19.5 108.2 – 0 0

100 2.0 58.0 60.8 106.3 0 10.5 105.2
200 2.1 43.6 94.7 101.4 0 16.9 170.7
250 2.0 41.1 92.9 110.8 0 17.1 155.5
300 2.1 42.3 104.2 90.0 1.9 19.1 165.0
350 2.0 39.1 107.0 84.1 136.2 19.8 165.5
400 2.1 38.8 99.6 64.7 223.2 20.3 130.3
677

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 www.soci.org
200 and 400 mmol L−1 . All the metabolites appeared from the
early stages and increased progressively during the fermentation
process.
The influence of initial glycerol/glucose molar ratio is more
evident for the consumption of glycerol (Table 1). The residual
glycerol concentration remained above 40% after 10 h of
incubation for initial glycerol concentration over 350 mmol L−1
while it dropped to nearly zero at lower concentrations. It was
also found that reuterin concentration reached a maximum
corresponding with the exponential cell growth phase in the
range of glycerol concentration investigated. A slight increase in
the reuterin concentration in the media was observed beyond
200 mmol L−1 glycerol concentration whereas the concentration
of 1,3-PDL was not affected by the initial glycerol/glucose molar
ratio over the range tested.
The product distribution indicates that glycerol competes
successfully with acetyl phosphate as a major hydrogen acceptor
during the fermentation of glucose, resulting in the accumulation
of 1,3-PDL, which is in accordance with the results obtained by
Talarico et al.29 for L. reuteri 1063 and Lüthi-Peng et al.26 for L. reuteri
(ATCC 53 608). In the presence of glycerol, the shift from ethanol
to acetate confirms the participation of glycerol in regeneration
of NAD+ by reduction of reuterin to 1,3-PDL. As long as NADH is
generated from glycolisis, reuterin is used as preferred electron
acceptor. The production of acetate instead of ethanol allows one
to obtain additional ATP, thus the need for producing ATP through
glycolysis is reduced. Additional ATP can be made by oxidizing
part of the intermediate pyruvate to acetate instead of the usual
reduction to lactate.30
Model development
In order to describe simultaneously the time evolution of biomass,
glucose and heterolactic fermentation products (lactate, acetate
and ethanol), a kinetic model has been proposed. The growth rate
of L. reuteri PRO 137 was described by means of a simplification of
Monod’s equation, which expresses the growth rate as a function
of substrate and biomass concentrations:
dX
= kx · S · X
dt
Glucose was the growth-limiting substrate in microbial growth
of L. reuteri PRO 137 in the MRS medium. The glucose consumption
rate can be expressed by an equation which takes into account
the yield of biomass on glucose:
dS 1 dX
− = · (2)
dt YXS dt
The production of lactate, acetate and ethanol, associated to
cell growth can be expressed by the following rate equations:
dL dX
= YLX ·
dt dt
dA dX
= YAX ·
dt dt
dE dX
= YEX ·
dt dt
Typical experimental values of microbial growth, substrate
consumption and product formation for a batch culture of L.
678
reuteri PRO 137 in glucose and the corresponding fitting curves
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c 2008 Society of Chemical Industry
M. Tobajas et al.
are shown in Fig. 1. The data were fitted to the proposed model
by a multi-response nonlinear regression method based on the
minimization of the sum of square residues. A reasonably good
agreement can be observed among the experimental results and
those predicted by the model. The values obtained for the kinetic
constants are summarized in Table 2.
An unstructured kinetic model has also been proposed to
describe simultaneously the variation of biomass, substrates
(glucose and glycerol) and products (lactate, acetate, ethanol,
reuterin and 1,3-PDL) concentrations during the cofermentation
of glycerol and glucose by L. reuteri. The kinetic model is based on
the previous equations for glucose consumption and formation
of the heterolactic fermentation products (Equations (2)–(5))
and the corresponding expressions for cell mass growth and
glycerol bioconversion. Bacterial growth is inhibited as the
reuterin concentration increases,1,25 which must be considered
in modelling cell growth rate. In this model, growth of L. reuteri
is described by an expression which takes into account the
evolution of biomass and substrate concentrations along the
process together with the toxic effect of reuterin accumulation for
L. reuteri:
dX X ·S
= k1 · (6)
dt k2 + R
Reuterin and 1,3-PDL production have been well described in
the literature by an intracellular series reaction pathway.25,26
Nevertheless, results showed that 1,3-PDL appeared at early stages
and increased progressively during the fermentation process,
which can be the overall result of a parallel/series reaction pathway,
i.e. 1,3-PDL could be produced by direct glycerol bioconversion and
by reduction of reuterin. Both are considered in the corresponding
equation for the production rate:
dP
= k4 · X · G + k5 · X · R (7)
dt
The rate of glycerol consumption is related to microbial growth
and production of reuterin and 1,3-PDL according to the following
expression:
(1) dG 1 dX
− = · + k3 · X · G + k4 · X · G (8)
dt YXG dt
Furthermore, reuterin produced by glycerol dehydration is
subsequently reduced to 1,3-PDL. All these reactions are taken
into account for the equation of reuterin production rate:
dR
= k3 · X · G − k5 · X · R (9)
dt
The experimental data were fitted to the proposed model
(Equations (2)–(9)). A multi-response nonlinear regression analysis
based on Marquardt algorithm combined with a Runge-Kutta
integration method at the 95% probability level was used to
(3)
obtain the values of the fitting parameters. The optimal-fitting sets
of parameter values are listed in Table 2.
(4)
The model predicts satisfactorily the evolution of biomass,
substrates and products concentrations for glycerol/glucose
(5) cofermentation by L. reuteri over a wide range of initial glycerol
concentrations. Figure 3 shows the experimental results for
microbial growth, substrates consumption and end-products
formation for different batch cultures of L. reuteri with initial
glycerol concentrations ranging from 100 to 300 mmol L−1 and
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Table 2. Fitting values of the parametersa of Equations (2)–(9) for the glycerol/glucose cofermentation by L. reuteri PRO 137 in MRS medium with
different glycerol concentration

Glycerol (mmol L−1 )

Parameter 0 100 200 250 300

kx 94.8 ± 2.5 – – – –
k1 – 1.51 ± 0.08 0.472 ± 0.301 0.149 ± 0.127 0.105 ± 0.051
k2 – 120 ± 45 43.6 ± 32.2 14.9 ± 13.8 8.39 ± 4.68
k3 – 0.290 ± 0.051 0.135 ± 0.058 0.0506 ± 0.0511 0.0595 ± 0.0342
k4 – 1.25 ± 0.50 0.298 ± 0.065 0.148 ± 0.052 0.109 ± 0.035
k5 – 0.423 ± 0.876 0.486 ± 0.301 0.458 ± 0.560 0.471 ± 0.303
YXS 222 ± 7 151 ± 8 217 ± 6 187 ± 9 216 ± 14
YXG – 131 ± 15 522 ± 89 228 ± 27 135 ± 11
YLX 45.5 ± 3.1 51.3 ± 5.5 39.6 ± 2.6 52.8 ± 4.2 45.3 ± 4.4
YAX 9.38 ± 2.80 31.6 ± 5.1 46.7 ± 2.7 41.1 ± 3.7 46.9 ± 4.4
YEX 41.7 ± 5.2 29.0 ± 3.9 20.7 ± 2.4 18.1 ± 3.5 21.2 ± 4.1
a Individual confidence limits at the 95% probability level.

(a) 180 glucose

2.5 (b) 240 glucose
2.5
lactate lactate
Concentration (mmol L-1)

Concentration (mmol L-1)

acetate acetate
150 ethanol
2.0 200 ethanol
2.0
biomass biomass
glycerol glycerol
Biomass (gL-1)

Biomass (gL-1)
reuterin reuterin
120 1,3-PDL 160 1,3-PDL
1.5 1.5
90 120
1.0 1.0
60 80

30 0.5 40 0.5

0 0.0 0 0.0
0 2 4 6 8 10 0 2 4 6 8 10 12
Time (h) Time (h)

(c) 280 glucose

2.5 (d) glucose
2.5
lactate 300 lactate
Concentration (mmol L-1)

Concentration (mmol L-1)

acetate acetate
240 ethanol ethanol
biomass 2.0 250 biomass 2.0
glycerol glycerol
Biomass (gL-1)

Biomass (gL-1)
200 reuterin reuterin
1,3-PDL 1,3-PDL
1.5 200 1.5
160
120 150
1.0 1.0
80 100
0.5 0.5
40 50

0 0.0 0 0.0
0 2 4 6 8 10 12 0 2 4 6 8 10 12
Time (h) Time (h)

Figure 3. Time-evolution of biomass, substrates and products along the cofermentation of 111 mmol L−1 glucose by L. reuteri PRO 137 with different
glycerol concentrations (a) 100 mmol L−1 , (b) 200 mmol L−1 , (c) 250 mmol L−1 and (d) 300 mmol L−1 . Experimental (symbols) and fitted (solid line) results.

111 mmol L−1 glucose. The corresponding curves showing the CONCLUSIONS
model predictions are also plotted. In general, the validity of Reuterin and 1,3-PDL were produced by L. reuteri from glyc-
the model can be concluded. Higher deviations were found erol/glucose cometabolism. Glucose was used as growth substrate
when working with initial glycerol concentrations beyond about and glycerol as an alternative hydrogen acceptor giving rise to
350 mmol L−1 (not shown in the figure). This fact could be reuterin and 1,3-PDL. However, a glycerol concentration higher
explained taking into account that glycerol remained in the than 100 mmol L−1 was necessary to produce reuterin and 1,3-
medium, i.e. glycerol was not limiting the process. When that PDL. Increasing the glycerol concentration from 200 to 400 mmol
occurs the system behaves in a different way, which cannot be L−1 has no significant effect on reuterin and 1,3-PDL production.
satisfactorily described by the proposed model. Nevertheless, Reuterin, which is an intermediate of the glycerol bioconversion,
these high concentrations of glycerol are not of interest since they reached the maximum concentration at 6 h and decreased with
679

do not improve reuterin and 1,3-PDL production. prolonged fermentation time since it is reduced to 1,3-PDL. The

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 www.soci.org
presence of glycerol did not affect biomass growth but altered the
end products profile by reducing ethanol formation and increas-
ing acetate production. Microbial growth, substrate consumption
and product formation during the growth of L. reuteri on glucose
alone or cofermented with glycerol were adequately described by
the kinetic model proposed.
ACKNOWLEDGEMENT
This study was supported by the Universidad Autónoma de
Madrid through the project no 1001020007.
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