Chemical Engineering Journal 305 (2016) 28–36

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Chemical Engineering Journal

journal homepage: www.elsevier.com/locate/cej

Production of lactic acid by ultrafiltration of fermented whey obtained

in bioreactor equipped with ZOSS membrane
Brygida Wojtyniak a,⇑, Joanna Kołodziejczyk a, Daniela Szaniawska b
a
West Pomeranian University of Technology, Faculty of Chemical Technology and Engineering, Al.Piastow 42, 70-065 Szczecin, Poland
b
Maritime University of Szczecin, Faculty of Economics and Transport Engineering, ul.H.Poboznego 11, 70-507 Szczecin, Poland

h i g h l i g h t s

Synthetic and industrial whey separation using ZOSS membrane is reported.

UF parameters, cross flow velocity, transmembrane pressure were investigated.

Rm and Ri were used to describe fouling mechanism.

Ri was described by exponential equation.

a r t i c l e i n f o a b s t r a c t

Article history: Experimental investigation on cross-flow and pressure ultrafiltration (UF) fermentation broth of lactic
Available online 3 February 2016 acid during the whey fermentation using a bioreactor equipped with zirconium oxide stainless steel
(ZOSS) membrane was carried out. Different filtration resistance (Rf and Ri) components were calculated
Keywords: to understand better the interactions between the membrane and fermentation feed. Various raw mate-
Ultrafiltration rials (deproteinated synthetic whey S1, model/synthetic whey S2 and ultrafiltration dairy whey permeate
ZOSS membrane S3) were used as a feed for UF tests under constant conditions. It was found that the highest concentra-
Whey
tion of lactic acid in permeate (0.69 ± 0.95%wt.) as well as permeate flux (0.5 ± 5.5  106 m3 m2 s1) was
Lactic acid
Lactobacillus acidophilus
obtained with the transmembrane pressure of 2  106 Pa and the cross-flow velocity of 1 m s1. Under
Fouling these conditions the fouling resistance of 6.87  1011 m1 was minimal. The integration of bioreactor with
ZOSS membranes seems to be promising concept in the production and filtration of lactic acid in the dairy
industry.
Ó 2016 Elsevier B.V. All rights reserved.

1. Introduction
A large amount of waste whey, rich in lactose, is generated in
the milk industry as a byproduct of cheese and casein production.
There is a continuing interest in identifying ways to transform this
byproduct into more marketable materials. Moreover, there is a
major commercial push to produce biodegradable plastics using
lactic acid derived from fermentation. It is also used for polymer-
ization to biodegradable polylactic acid (PLA), which is used for
medical applications such as sutures and clips for wound closure
or prosthetic devices. Therefore, there exists a challenge to identify
sources of waste lactose as substrates for production of bioprod-
ucts to satisfy a wide demand. The use of biotechnological tech-
niques to find the suitability of whey for lactic acid production
⇑ Corresponding author.
E-mail address: brygida.wojtyniak@gmail.com (B. Wojtyniak).
can serve dual purpose, i.e. production of valuable product, lactic
acid, thus increasing the economy of the process [1].
The conventional recovery processes of lactic acid from fermen-
tation broth are quite complicated. Separation from dilute wastew-
ater or fermentation broths using evaporation requires
vaporization of water, consuming much energy. Also distillation
is not useful due to non-volatility of lactic acid. In conventional
processes, precipitation of calcium lactate contains many steps:
precipitation, filtration, and addition of sulfuric acid, purification
using activated carbon, evaporation and crystallization. Thus, this
method of recovery is expensive and environmentally unfriendly
as it consumes lime and sulfuric acid and also produces a large
quantity of calcium sulfate sludge as solid waste, making these
separation methods expensive [2].
Bioprocessing or biotechnology is used in food production,
pharmaceuticals and chemistry with the aid of a biocatalyst, such
as an enzyme, microorganisms, plants cell or animal cell in a biore-
actor. The main step for bio-lactic acid production is fermentation,

http://dx.doi.org/10.1016/j.cej.2016.01.048
1385-8947/Ó 2016 Elsevier B.V. All rights reserved.
 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36
Symbols
C concentration [g dm3]
CFV cross-flow velocity [m s1]
DM dynamic membrane
Jv permeate flux, Eq. (1) [m3 m2 s1]
Jw clean water flux, [m3 m2 s1]
MRB membrane bioreactor
Rf fouling resistance [m1]
Ri resistance of the concentration polarization layer [m1]
Rm membrane resistance [m1],
RT total resistance [m1],
S0 acid dairy whey
S1 synthetic whey after removal of proteins by precipita-
tion and separation
S2 whey powder solution
S3 permeate after dairy whey ultrafiltration
t time [s]
T temperature [°C]
TMP trans membrane pressure
which runs in mild conditions but unluckily with rather low yield
[3]. As an alternative production method to fermentation, there is
chemical synthesis which is an energy intensive, high temperature
process far less selective, leading to additional costs for separation,
purification, and analytics. Thus, there is a necessity to develop
methods that raise both, process effectivity and product concentra-
tion. Membrane techniques have been shown as promising ways
for the lactic acid recovery. With advances in membrane tech-
niques, some processes, such as ultra- and microfiltration using
molecular sieves, could be hybridized with other technologies
and used as basic unit operations to concentrate lactic acid produc-
tion. Downstream processing to remove impurities, bulk-volume
reduction and simultaneous concentration of the desired product
from bioreactor are required [4]. Various steps developed during
downstream processing for recovering the finale product in lactic
acid production are illustrated in Fig. 1.
Pressure driven membrane techniques (PDMTs) based on inor-
ganic membranes and thermal swing adsorption using zeolites
are extremely useful for improving conventional bioprocesses
where separation of both raw materials and products is needed.
Membrane techniques are an attractive pursuit, mainly because of
their moderate operating parameters, chemical and physical stabil-
ity, and low energy consumption. On the other hand, the need to
sterilization during bioprocessing is a drawback [5]. Thus, inorganic
membranes are typically most suitable for these applications due to
their resistance to temperatures and pH fluctuations. Thus, mineral
membranes, particularly zirconium or titanium oxide supported by
carbon or by alumina and ceramic membranes are now being used
increasingly for ultrafiltration (UF), microfiltration (MF) and even
nanofiltration (NF) [6]. They are more expensive but have sustain-
ably longer life than membranes [7]. As fermentation broths usually
are diluted, contain many complex compounds, and also strongly
sensitive to operating conditions, membrane processes are well-
suited for downstream processing by filtration such MF, UF, NF
[8]. Membrane processes such as reverse osmosis (RO) and gas sep-
aration (GS) have already been demonstrated for upstream use of
the bioreactor [9]. The UF permeate contains lactose, non-protein
nitrogen and dissolved salts, and can be concentrated by RO to
the desired sugar concentration for fermentation [5,10].
Nowadays, the production of new environmental friendly prod-
ucts using renewables or waste seems to be very promising. Waste
sugars as lactose from whey can be gained as secondary raw mate-
rial during ethanol manufacturing (bio-fuel) or lactic acid
29
u cross-flow velocity [m s1]
V cumulative filtration volume, [m3]
Greek symbols
Dp transmembrane pressure [N m2]
l dynamic viscosity [Pa s]
Subscripts
b bacteria
i concentration polarization
m membrane
p permeate
r retentate
f fouling
L lactose
LA lactic acid
w water
(biodegradable polymers) using fermentation. The distinctive form
of bioprocesses necessitates precise separation of desired products
from multicomponent, and usually diluted, systems. Here separa-
tion problems are unavoidable [1,11].
A key hurdle in the course of an efficient and environmentally
compatible hybrid process for producing lactic acid by fermenta-
tion is the lack of a simple and effective method for maintaining
a pH that is permissive to bacteria. Dynamic membrane (DM) tech-
nique is a novel means of membrane modification. DM technology
may be a promising approach to solve problems encountered in
membrane bioreactor (MBR) process, as the membrane itself may
become redundant, since solids rejection is accomplished by the
secondary membrane layer which can be formed in situ as a self-
assembling dynamic membrane (SFDM). Periodic formation and
removal of DMs may reduce membrane permeability losses as
encountered in conventional MBRs. Their reliability has already
been demonstrated in several MBR applications, such as treatment
of oily wastewater treatment [12] and anaerobic biodegradation
[13].
The main purpose of this study was to determine operating
parameters to produce lactic acid by ultrafiltration of fermented
whey obtained in bioreactor equipped with ZOSS membrane. Dif-
ferent operating parameters have been proposed varying the trans-
membrane pressure (TMP), cross-flow velocity (CFV) and
composition of fermentation broth (acid whey from dairy industry,
S0, deproteinated synthetic whey, S1, model/synthetic whey, S2,
permeate from dairy whey ultrafiltration, S3, lactic acid bacteria,
LAB and supplements). In the experimental study, the TMP (1–
2.5  106 Pa), CFV (0.5–2.6 m s1), LAB (0.07–1.0 g dm3) content
and supplements addition have been varied to determine their
effects on permeate flux, Jv, m3 m2 s1 and lactic acid concentra-
tion in permeate, CLA; g dm3.
2. Theory
2.1. Cross-flow ultrafiltration
Ultrafiltration (UF) is characterized by membrane pore size in
range between 0.05 and 2 lm and operating pressure between 1
and 30 bar [14]. The general terms that are used in the description
of membrane separation process, such as ultrafiltration are the
permeate flux, (Jv) and rejection (r). The permeate flux can be
described by Eq. (1)
30 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36
Fig. 1. Bioprocessing by membrane-based hybrid separation processes (a) process flow diagram from commercial whey processing (b) integrated membrane system for
continuous lactic acid production with whey treatment.
Dp
Jv ¼
lRT
in which, Jv is permeate flux m s1, Dp is effective transmembrane
pressure N m2, l is dynamic viscosity Pa s, RT is total resistance
of the membrane m1.
The membrane exhibits rejection for the solute in the feed at a
concentration level CF, so partial permeation will occur and
rejected solute accumulates in the boundary layer due to concen-
tration polarization phenomenon. At the feed and permeate sides
of the membrane will occur solute distribution processes, by which
a final solute concentration in permeate is reached at the level of
CP. In membrane processes solute behavior is given by rejection
coefficient, r (Eq. (2)):
Cp
r ¼1
Cr
2.2. Fouling analysis
The ultrafiltration of solutions containing biological substances,
such as proteins and microorganisms, causes fouling occurs pre-
dominantly on the membrane surface. It occurs through one or
more various mechanisms and leads to flux decline. The flux
decline can be attributed to concentration polarization, adsorp-
ð1Þ
tion/deposition of compounds being separated within the mem-
brane structure, pore blockage and cake formation. Flux decline
due to concentration polarization is reversible; it can be removed
by changing process parameter or by rinsing with water. Flux
decline due to cake formation is very often irreversible and
requires chemical cleaning. Fouling caused by adsorption and pore
blockage within the membrane is usually irreversible, although
fouling due to pore blockage could be partially reversible depend-
ing [15]. Permeate flux, Jv, which is defined as flow per membrane
unit area, A, is directly proportional to the driving force, i.e. trans-
membrane pressure, Dp and inversely proportional to the total
resistance, RT (Eq. (3)):
1 dV TMP
Jv ¼  ¼ ð3Þ
ð2Þ A dt lRT
where A is area of membrane m2, t is filtration time s, V is permeate
volume m3, Dp is pressure difference across the membrane Pa.
Fouling of membrane by different fouling mechanisms can be
described by resistance-in-series model. In this model, the total
resistance of the membrane, RT is composed of three resistances
in series (Eq. (4)):
RT ¼ Rm þ Ri þ Rf ð4Þ
 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36 31

where, Rm – resistance of clean membrane to permeation of deionized The fermentation medium was sterilized at 92 °C on 3 consecu-
water; this resistance is time independent; Ri – initial fouling resis- tive days, for 30 min each day. Lactobacillus acidophilus (Biolacta,
tance, caused by concentration polarization phenomena and fouling Olsztyn, Poland) was used for the lactic acid fermentation. Bacteria
of the membrane during startup of the process; this resistance is also were cultured in Ringer’s medium. The whey was supplemented
independent of time; Rf – time dependent fouling resistance. with components of broth, consisting of 10 g of yeast extract,
The extent of membrane resistance depends on the membrane 0.5 g of NH4NO3 and 5 cm3 of Tween 80 (polysorbit-80) per liter
thickness and various morphological features such as the tortuos- of distilled water.
ity, porosity, and pore size distribution. Thus, the total resistance to Lactic acid was measured by high-performance liquid chro-
flow can be described by two types of resistances, time-dependent, matography with ion exclusion column (NucleogelR Ion 300 0A,
which increases with time, and time-independent, which remains Macherey-Nagel) by detection of the refractive index. The mobile
constant. phase was 0.01 N H2SO4 solution at a flow-rate of 0.6 ml/min.
The initial fouling resistant, Ri can be correlated empirically The temperature of the detector and of the column was maintained
with operating parameters, i.e. transmembrane pressure and cross at 30 °C and 35 °C, respectively. Lactose concentration was deter-
flow velocity as well as feed properties, compound concentration mined by the Bertrand method [17].
and pH (Eq. (5)) [16]. Experimental investigations were carried out for continuous
mode for fermentation and purification with pH adjustment. Typ-
Ri ¼ a  ub  C cb  Dpd  pHe ð5Þ
ical operating conditions were between pH 4 and 5; pH of the sam-
a, b, c, d and e are empirical constants. ples was measured with a pH meter CP-411 (Elmetron).
In general, fermentation broths contain proteins, bacterial cells,
colloidal particles and salts, so that each of these fouling mecha-
3.2. Cross-flow ultrafiltration tests
nisms may occur (protein adsorption, bacterial attachment, depo-
sition of colloidal particles and salts precipitation).
Class 2 (precoated) ZOSS membranes have been used during the
Calculations are essential in the design and operation of mem-
ultrafiltration tests, which were obtained by deposition of inor-
brane processes. Specifically, it is of paramount importance to be
ganic colloids (oxides, hydroxides and salts of zirconium, alu-
able to calculate all the relevant fundamental properties of the sub-
minum and iron) on selective microporous supports. In this case
stances of interest in all required process conditions. The model
the hydrous Zr(IV) oxide is used as a dynamic layer deposited by
contains four parameters (u; Cb; Dp; pH) which can be used to fit
filtration of the solution under constant pH (4.1) and pressure
the model to the data for any given situation. The total resistance
(5.2  106 Pa) through the tubular stainless steel support with
(RT) was calculated with constant filtrate flow rate using Eq. (3).
diameter of 16 mm and surface of 0.029 m2 covered with TiO2
The resistance (Ri) caused by initial fouling was estimated accord-
layer. The membrane was characterized by water permeability at
ing to Eq. (5), it can be seen that the resistance Ri is estimated with
the level of 144  105 m3 m2 s1. The used membranes were
experimental data. A model was developed in which non-linear
washed according to the procedure with 0.01 M HCl solution at a
factors were determined by using the program ‘‘STATISTICA”.
pressure of 1–5  106 Pa [18] and formed once again.
Filtration tests were performed using an UF pilot plant previ-
3. Experimental ously described in [19]. Before each run, the membrane unit was
cleaned with sterile water and sterilized with Neoseptal D (Dr.
3.1. Feed preparation Weigert) 1%wt. solution. Temperature was set at 30 °C, TMP was
1.0  106 Pa and CFV was 2.0 m s1. The total recirculation mode
The source of lactose used in this investigation and their was followed for 30 min.
characteristics are described in Table 1. The tests were divided into two types of measurements: UF and
fermentation coupled with UF tests. Fermentation took place in a
feed tank with a volume of 20 dm3 under controlled temperature.
Table 1
Raw material characterization. UF tests of model whey solution in the feed tank were performed
with recirculation of the retentate. Retentate was recycled using
Symbol Characteristics
the membrane module and returned to the feed tank through the
S1 Synthetic whey after removal of Synthetic whey was prepared from pump and heat exchanger. After filtration the permeate was col-
proteins by precipitation and whey concentrate; the composition lected in a vessel. The working volume of reactor was maintained
separation; lactose content, of whey concentrate: protein (Biuret)
27%wt., lactose 65%wt., nitrogen 2%
at a constant value by supplying feeding medium in balance with
CL = 0.172%wt. by weight
wt., moisture 4.4%wt., the process of permeate flow of membrane. After 12 h about 3.5 dm3 of permeate
deproteinization was carried out at was collected. The operating parameters of the tests and its results
95–98 °C and pH 4.6 for 30 min., next are presented in Table 2.
precipitated proteins were separated
by precipitation
S2 Synthetic whey prepared from Synthetic whey solution was 4. Results and discussion
whey concentrate; lactose prepared by dissolving 77.1 g of whey
content, CL = 2.5%wt. by weight concentrate (Sigma Aldrich) into
4.1. Ultrafiltration of fermented synthetic whey obtained in bioreactor
20 dm3 of deionized water (DI) and
stirring for 5 min at ambient equipped with ZOSS membrane
temperature
S0 Acid whey from local District Acid whey composition: 4.93%wt. The aim this study was examination of the influence of selected
Dairy Cooperative (Poland); lactose, pH = 4.95; total protein 0.8– operating parameters on the production of lactic acid by lactose
lactose content, CL = 4.93%wt. by 1.0%wt., ash 0.5%wt., fat 0.2%wt., fermentation in a bioreactor equipped with a ZOSS membrane.
weight sodium, calcium, potassium 0.03%wt. The fermentation process was performed at controlled tempera-
and water 93.5–95%wt.
ture, transmembrane pressure and cross flow velocity (T = 37 °C;
S3 Industrial whey ultrafiltration Industrial whey was filtered by Dp = 1.0  106 Pa and u = 1.0 m s1). Fig. 2 shows profiles of perme-
permeate; lactose content, membrane ZOSS for separation of
ate flux and lactic acid concentration in permeate vs time obtained
CL = 3.83%wt. by weight proteins
during first step of experimental study.
32 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36

Table 2
Steps of experimental study using ZOSS membrane.

Research Feed/fermentation broth UF conditions Results

step
1 Fermentation broth obtained using Dp = 1.0  106 Pa; u = 1.0 m s1; Cb = 0.07 g dm3; T = 37 °C; pH = 4.0; CpLA = 0.083%wt.
deproteinated synthetic whey, S1 + La. pH = 4.6; pH = 5.0
Acidophilus
2 Fermentation broth obtained using synthetic Dp = 1.0  106 Pa; u = 0.5 m s1; Cb = 1.0 g dm3; 10 g dm3 of yeast CpLA = 0.76%wt.;
whey, S2 + La. Acidophilus and nutrients extract, 0.5 g dm3 of NH4NO3, and 5 g dm3 of Tween-80; T = 37 °C Dp = 1  106 Pa;
pH = 4.0; pH = 4.6; pH = 5.0 u = 0.5 m s1; pH = 4.6
Dp = 1.0–2.0  106 Pa; u = 1.0–2.6 m s1; Cb = 1.0 g dm3; 10 g dm3 CNH4 NO3 = 0.000603 g dm3
of yeast extract, 0.5 g dm3 of NH4NO3, and 5 g dm3 of Tween-80; CpLA = 0.16%wt.;
T = 37 °C; pH = 4.6 Dp = 1  106 Pa;
u = 2.6 m s1
CNH4 NO3 = 0.0007 g dm3
CpLA = 0.54%wt.;
Dp = 2  106 Pa;
u = 2.6 m s1
CNH4 NO3 = 0.0006 g dm3
3 Feed – acid whey from local District Dairy Dp = 1.3  106 Pa; u = 1.4 m s1 CpL = 4.84%wt.; pH = 4.86
Cooperative (Poland); S0 Dp = 2.3  106 Pa; u = 1.4 m s1 CpL = 4.86%wt.; pH = 4.54
Dp = 2.5  106 Pa; u = 1.9 m s1 CpL = 3.83%wt.; pH = 4.52
pH = 4.5; T = 20 °C
4 Fermentation broth obtained using permeate Dp = 2.0  106 Pa; CL = 1.2%wt.;
after dairy whey ultrafiltration, S3 + La. u = 1.0 m s1 CpLA = 0.98%wt.;
Acidophilus and nutrients Cb = 1 g dm3; 10 g dm3 of yeast extract, 0.5 g dm3 of NH4NO3, and CrLA = 0.25%wt.;
5 g dm3 of Tween-80; T = 37 °C; pH = 4.96 CNH4 NO3 = 0.0019 g dm3

10 g dm3 of yeast extract, 0.5 g dm3 of NH4NO3 and 5 g dm3

of Tween-80.
The effects of cross-flow velocity, transmembrane pressure, and
feed concentration on the flux and rejection of lactic acid in the
total recycle filtration were studied with UF membrane which is
described elsewhere [19]. The transmission of the lactic acid mole-
cules from the feed to the permeate side depends on the rejection
characteristics of the membrane and the concentration polariza-
tion layer.
The results presented in the Fig. 3 show four profiles of lactic
acid concentration in permeate vs time under varying fermenta-
tion conditions. Compared to the runs under different cross flow
velocity, the lactic acid productivity was initially low and then it
continuously increased. It was observed that the lactic acid con-
centration in permeates separated from fermentation broth is
higher at low operating parameters, i.e. transmembrane pressure
of 1 MPa and cross flow velocity of 0.5 m s1. The final lactic acid
Fig. 2. Lactic acid concentration and permeate flux vs ultrafiltration time of
fermented whey, S1 by ZOSS membrane at T = 37 °C; CFV = 1.0 m s1,
TMP = 1  106 Pa, Cb = 0.07 g dm3.

It can be seen from Fig. 2 that lactic acid concentration in per-

meate increases with fermentation time. On the contrary permeate
flux decreases with the increase of fermentation time, which is
caused by fouling. The highest lactic acid concentration in perme-
ate at the level of 0.083%wt. by weight was obtained after 10 h of
fermentation. The lowering in lactic acid concentrations after a
period of 10 h is probably due to the inhibitory action of the pro-
duct on the fermentation process, but may also be due to the low
lactose and bacteria concentration in feedstock and lack of nutri-
ents. In the case of growth of lactic acid shows an irregular expo-
nential curve. The permeate flux decline during time, which may
be caused by due to the plugging of pores by the broth fermenta-
tion, although small amounts of several substances.
In order to compare the ZOSS membrane performance and
establish membrane parameters additional experiments with syn-
thetic whey were carried out. The next experiments (step 2) were
performed with addition of nutrients. Moreover, this step of exper-
Fig. 3. Ultrafiltration of fermented synthetic whey (S2) by ZOSS membrane at 37 °C,
imental study was carried out with the aim to analyze effect of constant LAB concentration, Cb = 1.0 g dm3 and nutrients. Influence of cross-flow
TMP (1–2  106 Pa) and CFV (0.5–2.6 m s1) on lactic acid concen- velocity 0.5–2.6 m s1; pH = 4.6 and transmembrane pressure 1–2  106 Pa on lactic
tration – time profiles. The model whey was supplemented with acid production.
 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36 33

concentration reached a maximum value of 0.76%wt. by weight

after 14 h.

4.2. Ultrafiltration of fermented industrial whey obtained in bioreactor

equipped with ZOSS membrane

Step 3 in this study was the examination of the membranés

behavior during ultrafiltration of sour whey from dairy as a func-
tion of time (0–60 min), transmembrane pressure, TMP (1.3–
2.5  106 Pa) and cross flow velocities, u (1.4 and 1.9 m s1). The
selected results of permeate flux vs time are presented in Table 3
and Fig. 4.
The hydraulic permeability of the membrane is a very impor-
tant parameter from practical point of view, as a high permeate
flux (Jv) and small membrane area are needed to achieve high per-
formance in industrial scale plants. The results summarized in
Table 3 illustrate that permeate flux generally increases with
increasing transmembrane pressure and cross-flow velocity in
time. As it can be seen from Table 3 and Fig. 4 growth of studied Fig. 4. Effect of time, transmembrane pressure and cross-flow velocity on permeate
flux during ultrafiltration of sour dairy whey, S0 by ZOSS membrane at pH = 4.5 and
operating parameters, Dp and u leads to permeate flux increasing
T = 20 °C.
by 50%.
In line with theoretical expectations ZOSS membrane perme-
ability increases with increased transmembrane pressure and
cross-flow velocity in analyzed range of parameters. The analysis
of Fig.4 also shows that the impact of transmembrane pressure
on Jv is growing stronger for higher CFV of 1.9 m s1. The data
obtained in step 3 was used for fouling analysis presented in
Section 4.3.
The permeate obtained by ultrafiltration of acid dairy whey was
used for preparation of fermentation broth applied in experiments
conducted in step 4. Fig. 5 presents changes in the permeate flux
during the consecutive deproteinated whey as the functions of
the pressure for the membrane performance. As expected, flux
grows with increasing pressure and finally reaches a limiting value
indicating pressure filtration, probably caused by the blocking of
proteins on the membrane surface. Up to TMP of 2.5  106 Pa and
1.9 m s1, the limiting value of flux was not yet reached.
In step 4 the ultrafiltration process was performed using perme-
ate after ultrafiltration whey (S3) supplemented by LAB
Fig. 5. Ultrafiltration of dairy whey, S3 by ZOSS membrane at 37 °C, Cb = 1 g dm3,
(Lactobacillus acidophilus) with a concentration of 1 g dm3 and TMP = 2  106 Pa, CFV = 1 m s1 lactic acid concentration and permeate flux vs time.
selected nutrients at a cross-flow velocity of 1 m s1 and
transmembrane pressure of 2  106 Pa. As nutrients, yeast extract,
NH4NO3 and Tween-80 were used. The initial temperature of the Table 4
feed was 37 °C. The pH of the original acid whey was 4.96. Fig. 5 Deionized water flux through ZOSS membrane vs transmembrane pressure at 20 °C.
shows a typical result characterizing the investigated process, Dp 106 [Pa] Jw 104 [m s1]
i.e. profiles of lactic acid concentration and permeate flux vs time.
0.5 0.643
These experiments were carried out with LAB and nutrient 1.0 1.29
supplementation (Table 2). As is apparent from Fig. 5 after 12 h 2.0 2.57
of fermentation lactic acid concentration in permeate reached a 2.5 3.22
value of 0.98%wt. and the lactic acid production efficiency
increased to 64%, the resulting dependence is given by Eq. (2). In
the case of continuous separation and removal of lactic acid in the membrane reactor system, the broth concentration comprises
an important parameter affecting the magnitude of permeate flux.
Table 3
Selected results of sour dairy whey ultrafiltration by ZOSS membrane at pH = 4.5 and
4.3. Fouling analysis
T = 37 °C.

CFV [m s1] TMP 106 [Pa] t [s] Jv 106 [m3 m2 s1] As described above, total resistance, RT for membrane systems
1.4 1.3 ± 0.1 1800 2.29 consists of three resistances in series (Eq. (4)).
2700 1.80 The model was chosen as a first step of investigation in viewing
3600 1.71 the effect of microorganisms on filtration performance. Indeed, this
1.4 2.4 ± 0.1 1800 3.78
2700 3.70
paper focuses on the performance of ultrafiltration ZOSS mem-
3600 3.69 brane and the properties of the fouling by fermentations broth.
1.9 2.4 ± 0.1 1800 3.83 Special attention was paid to the hydraulic resistance of the initial
2700 3.78 resistance of the fouling. The impact of a yeast/bacteria mixture
3600 3.76
was experienced and discussed.
34 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36

4.3.1. Membrane resistance

Fig. 6. Effect of transmembrane pressure on water flux through ZOSS membrane.
Membrane resistance, Rm, was calculated from pure water flux
experiment through Hagen–Poiseuille equation:
Dp
Jw ¼ ð6Þ
l w  Rm
where Jw represents pure water flux and lw represents water vis-
cosity: 8.94  104 Pa s. The pure water was filtered through the
membrane and the pressure was measured by a pressure gauge.
Table 4 shows the change in flux with increase of transmembrane
pressure. This is done in order to calculate the membrane resistance
using deionized water. The experiments were run over period of
20 min by temperature 20 °C. It can be seen that increase in the
pressure across the membrane increases the flux.
Fig. 6 shows a graph of Jw vs Dp where the slope of the line can
be used to calculate the resistance of the given membrane.
Using experimental data straight line equation was determined
with correlation coefficient, R2 = 1. The membrane resistance was
found to be 8.69  1010 m1.

Fig. 7. (a) Effect of pH and cross flow velocity on resistance Ri and (b) effect of pH and permeate flux (Jv) on resistance (Ri) for Dp = 1.0–2.0  106 Pa; Cb = 1.0 g dm3;
fermentation broth (S2).
 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36 35

4.3.2. The initial fouling resistance, Ri

The dependence of resistance, Ri caused by concentration polar-
ization phenomenon and fouling of the membrane during startup
of the process on operating parameters was calculated using Eq.
(5) and ‘‘STATISTICA” by non-linear estimation. Few studies have
examined systematically the effects of cross flow velocity, pH
and bacteria concentration on the Ri. The resulting dependence is
given by Eq. (7):

Ri ¼ 27; 05  108  u2;1  C 1;7

b  Dp
1;0
 pH4;3
Exponential exponents in these equations (c), (d) and (e) are
positive, what indicates that increasing these operating parameters
lets resistance Ri grow. The largest positive impact on Ri is pH. The
impact of cross flow velocity on Ri is negative (b < 0). The effect of
pH and u on Ri during ultrafiltration is presented in Fig. 7.
The results given above are based on the correlation and autho-
rize to conclude that the initial fouling resistance exhibited higher
values than that of the membrane resistance. It is clearly indicated
that the solutes were probability adsorbed to the pores surfaces of
the ZOSS membrane.
In Fig. 8 the influence of pH and permeate flux (Jv) on resistance
(Ri) during ultrafiltration of fermentation broth (S1) is shown.
The profile suggests that the pH effects are profound, as evident
in the values of the permeate fluxes. The solutions’ pH affected the
membrane performance gradually, initial fouling resistance (Ri)
grows and membrane permeability (JV) decreases with increasing
pH (Fig. 8).
It is evident from Eq. (7) and Fig. 8 that pH values in the narrow
range of 4.4–4.7 are recommended for the separation of lactic acid
solution from fermentation broth.
4.3.3. Fouling resistance, Rf
The resistance due to fouling, Rf was evaluated using experi-
mental data and calculations by applying StatgraphicsÒ Centurion
XVI. The fouling resistance, Rf is considered to be proportional to
the amount of matter deposited on the membrane, which increases
with time until reaching a value of equilibrium, where the deposi-
ð7Þ
Fig. 9. Dependence of RT, Ri and Rf resistances on operating parameters (Cb, u, Dp).
tion of foulants is balanced by the tangential migration of particles
dragged by the retentate flow [20].
The resistance Rf was evaluated using Eqs. (3–5). Selected
results of the calculations are presented in Table 5 and Fig. 9.
According to the graph the resistance values vary with operation
parameters such as transmembrane pressure, cross flow velocity
and bacteria concentration during the entire fermentation course
in the membrane bioreactor.
The active membrane layer resistance, Rm and resistance of con-
centration polarization and initial fouling, Ri depends strongly on
dynamic process conditions, especially on cross-flow velocity.
Analysis authorizes the graph to find that the ratio of substances
contained in the feed has studied the effect of operating parame-
ters on the resistance due to fouling. Its value is the highest at low-
est investigated cross flow velocity u = 0.5 m s1, decreases with
increasing speed and pressure, and increases with increasing con-
centration of bacteria S1 and S3.

Fig. 8. Effect of pH and permeate flux (Jv) on resistance (Ri) for Dp = 2.0  106 Pa; u = 1.0 m s1; Cb = 0.07 g dm3; fermentation broth (S1).

Table 5
Effect of operating parameters on fouling resistances (RT, Ri, Rf) characterizing investigated systems.

System Cb [g dm3] u [m s1] Dp 106 [Pa] RT 1012 [m1] Ri 1012 [m1] Rf 1012 [m1]
S1 0.07 1 1 1.83 0.574 1.7
S2 1 2.6 1 1.28 0.736 5.45
2.6 2 0.729 0.250 4.79
0.5 1 34.4 24.7 9.70
S3 1 1 2 7.82 7.16 0.687
36 B. Wojtyniak et al. / Chemical Engineering Journal 305 (2016) 28–36
Experimental results summarized in Table 5 and Fig. 9 confirm
that changes in analyzed operating parameters (Cb, u, Dp) have sig-
nificant effects on the resistances RT, Ri and Rf. The highest values of
these resistances were observed for u = 0.5 m s1 and TMP
1.0  106 Pa. The increase of cross-flow velocity and transmem-
brane pressure to 2.6 m s1 and 2.0  106 Pa, respectively reduces
values of all resistances.
However, with the variation of flux the transport rate of prod-
ucts to the membrane correlates, so the observed fouling behavior
is not solely the result of membrane/foulant interactions. In most
cases permeate flux is considered a key design parameter for mem-
brane systems and reflecting its productivity. The two factors that
led to deterioration of the flux rate were fouling and concentration
polarization.
5. Conclusion
This study has demonstrated that ZOSS membrane integrated in
a fermentation process could separate product of fermentation
from fermentation broth. The experimental results showed that
the process and operability design of an integrated bioreactor
and membrane separation process is usable in ultrafiltration in
downstream processing. Special attention was paid on lactic acid
concentration in permeates and flux decline due to three resis-
tances, i.e. membrane resistance, Rm, initial resistance, Ri and foul-
ing resistance, Rf.
Based on the results and calculations presented in this study
and results obtained in [19] it can be concluded that the lactic acid
concentration (CLA) in permeates separated from fermentation
broth, synthetic whey prepared from whey concentrate (S2) is
higher at low operating parameters, i.e. transmembrane pressure
of 1.0  106 Pa and cross flow velocity of 0.5 m s1 and the flux
increases with increasing transmembrane pressure. The final lactic
acid concentration reached a maximum value of 0.76%wt. by
weight after 14 h of fermentation. The increase of cross flow veloc-
ity and transmembrane pressure to 2.6 ms1 and 2.0  106 Pa,
respectively reduces values of all resistances to minimum. The
optimum transmembrane pressure can be suggested to be approx.
2.0  106 Pa where the fouling is minimal. On the other hand, the
values of lactic acid concentration were of 0.54%wt. Note that the
permeate concentration obtained can strongly depend on the val-
ues of the separation factor as was showed in Fig. 5.
Higher cross flow rates can reduce membrane fouling by pro-
viding a shear force to sweep away deposited materials. At high
feed flow rates, more lactic acid (CLA) were led by the membrane
with less sugar (CL) permeating through. The optimum feed flow
rate was 1.0 m s1. The ultrafiltration process can effectivity
increased distinctively by continuous removal of product solution
from fermentation broth.
Future studies may be directed toward more experiments with
industrial whey as feedstock as the present investigation was
carried out mainly using synthetic whey which that indicated the
direction of the research.
Acknowledgement
The research was partially financially supported by the Polish
Ministry of Science and higher Education (grant No. 3/S/IIT/14).
References
[1] B. Wojtyniak, E. Gabrus, D. Szaniawska, Conversion of waste lactose to ethanol
and lactic acid using hybrid biomembrane-adsorption processes, in: Membr.
Sci. Technol. Conf. PERMEA 2003, Tatranské Matliare, Slovakia, 2003.
[2] K.L. Wasewar, Separation of lactic acid: recent advances, Chem. Biochem. Eng.
Q. 19 (2005) 159–172.
[3] Y. Wang, Y. Tashiro, S. Kenji, Fermentative production of lactic acid from
renewable materials: recent achievements, prospects, and limits, J. Biosci.
Bioeng. 119 (2015) 10–18.
[4] P. Börgardts, W. Krischke, W. Trösch, H. Brunner, Integrated bioprocess for the
economic production of lactic acid from whey permeate, Bioprocess. Eng. 19
(1998) 321–329.
[5] A. Hinkova, P. Zidova, V. Pour, Z. Bubnik, S. Henke, A. Salova, P. Kadlec, Potential
of membrane separation processes in cheese whey fractionation and
separation, Procedia Eng. 42 (2012) 1425–1436.
[6] J. Sikder, S. Chakraborty, P. Pal, E. Driol, Purification of lactic acid from
microfiltrate fermentation broth by cross-flow, Biochem. Eng. J. 69 (2012)
130–139.
[7] C. Fox, P.L. McSweeney, T.M. Cogan, T.P. Guinee, Cheese: Chemistry, Physics
and Microbiology: General Aspects, Academic Press, 2004.
[8] J. Sikdera, M. Roy, P. Dey, P. Pal, Techno-economic analysis of a membrane-
integrated bioreactor system for production of lactic acid from sugarcane juice,
Biochem. Eng. J. 63 (2012) 81–87.
[9] R. Singh, Hybrid Membrane Systems for Water Purification, Elsevier, 2005.
[10] Y. Li, A. Shahbazi, K. Williams, C. Wan, Separate and concentrate lactic acid
using combination of nanofiltration and reverse osmosis membranes, Appl.
Biochem. Biotechnol. 147 (2008) 1–9.
[11] F. Salehi, Current and future applications for nanofiltration technology in the
food processing, Food Bioprod. Process. 92 (2014) 161–177.
[12] T. Yang, Z.-F. Ma, Q.-Y. Yang, Formation and performance of Kaolin/MnO2 bi-
layer composite dynamic membrane for oily wastewater treatment: effect of
solution conditions, Desalination 270 (2011) 50–56.
[13] M. Ersahin, H. Ozgun, Y. Tao, J. van Lier, Applicability of dynamic membrane
technology in anaerobic membrane bioreactors, Water Res. (2014) 420–429.
[14] A. Nordin, A. Jönsson, Influence of module configuration on total economics
during ultrafiltration at high concentration, Chem. Eng. Res. Des. 88 (2010)
1555–1562.
[15] B. Tensel, W. Bao, I. Tansel, Characterization of fouling kinetics in
ultrafiltration systems by resistances in series model, Desalination 129
(2000) 7–14.
[16] A. van Boxtel, Strategies for optimal control of membrane fouling: reverse
osmosis of cheese whey (a case study, Ph.D. thesis), Twente University of
Technology, Enschede, The Netherlands, 1991.
[17] Milk and milk products -Whey -Methods of analysis, Polish norm PN-A-86364,
1996.
[18] S. James, J. Johnson, Based on the Symposium on Polymers for Desalination,
Held at the 162nd National Meeting of the American Chemical, Research,
Reverse Osmosis Membrane, Society in Washington, D.C. 1972, p. 394
[19] B. Wojtyniak, D. Szaniawska, Separation of lactic acid solutions from whey
fermentation broth using zirconium (IV) hydrous oxide dynamically formed
membrane, Pol. J. Environ. Stud. 24 (3) (2015) 1387–1393.
[20] R. Perez-Galvez, E.M. Guadix, J.-P. Berge, A. Guadix, Operation and cleaning of
ceramic membranes fpr the filtration of fish press liquor, J. Membr. Science
384 (2011) 142–148.