J Process Biochemistry Vol. 33, No. 2, pp.

199-203, 1998
;,N"x ¢7 il © 1998 Elsevier Science Ltd
• " f -
All rights reserved. Printed in Great Britain
01)32-9592/98 $19.0t) + 0.00
ELSEVIER
PII: S0032-9592(97)00065-4

Surfactants in anaerobic digestion of salty

cheese whey using upflow fixed film reactor for
improved biomethanation

Priti Patel and Datta Madamwar*

Post Graduate Department of Biosciences, Sardar Patel University, Vallabh Vidyanagar 388 120, Gujarat, India

(Received 3 January 1997; revised version received 5 July 1097; accepted 5 July 1997)

Abstract

In order to improve the anaerobic digestion process of salty cheese whey, the effects of various doses of
surfactants: Tween 80, Triton-X-100, Tegopren 3022, sodium lauryl sulphate and Cetrimide, have been
studied in an upflow fixed film bioreactor. Among these, sodium lauryl sulphate showed a 70% increase in
gas production with a higher methane content (77%) and improved biodegradation. © 1998 Elsevier Science
Ltd
Keywords:biomethanation, fixed film reactor, salty cheese whey, surfactant, energy, anaerobic digestion•

Introduction process stability. Among the advanced technologies of

There has always been a great deal of interest in anae-
robic fermentation of industrial waste [1,2]. A large
number of dairies in India dispose of their waste,
especially cheese whey, into the environment in enor-
mous quantities. Cheese whey, a by-product of cheese
making, has a high organic content with a chemical
oxygen demand (COD) of about 60-80 g litre- ~, which
leads to disposal problems. Anaerobic digestion of
cheese whey offers a positive environmental impact
since it combines waste stabilization with net fuel pro-
duction and allows the use of the effluent as fertilizer.
The major advantages of this process are low cost, high
energy efficiency and process simplicity as compared to
other waste treatment methods. However, in spite of
waste reduction and energy potential, anaerobic diges-
tion is not a popular process in the dairy industry. This
is largely due to the problem of slow reaction rates
which require long hydraulic retention time (HRT) and
poor process stability using conventional reactors. The
low growth rate of anaerobic micro-organisms has
encouraged the development of high rate anaerobic
bioreactors, in order to avoid the loss of micro-organ-
isms in the effluent stream and to provide better
*To whom correspondence should be addressed.
199
anaerobic digestion, the fixed film reactor allows effi-
cient digestion of high and low strength soluble waste
at shorter H R T [3,4]. Our previous study has shown
improved gas production with enriched methane
content when salty cheese whey [with salt concentra-
tions from 4 to 6% (w/v)] was used in a fixed film
reactor at HRT of 2 days using charcoal as bedding
material [5].
There is further growing interest in maximizing
methane production from salty cheese whey, especially
those with high Na + concentrations. The presence of
high concentrations of sodium ions is detrimental to
the performance of anaerobic bioreactors [6], and this
problem was overcome by diluting salty cheese whey
with total dairy waste with a low total solid content and
by developing salt tolerant inocula. The mixing of total
dairy waste with salty cheese whey reduces the salt
content of the whey and supports the growth of salt
tolerant methanogens. This dilution also provided an
operational advantage due to the reduction in total
solids. It has also been reported that surfactants
increased the productivity of anaerobic digestion
systems [7]. They show unusual catalysis of organic
reactions and are known for the enhancement of bio-
degradation of organic compounds [8-10]. An attempt
has been made to determine the efffect of various sur-
200 P Patel & D. Madamwar
factants on anaerobic digestion of salty cheese whey
using fixed film reactors.
Materials and methods
All chemicals used were of analytical grade. Cheese
whey and dairy waste-water were collected from
AMUL dairy (Cheese Manufacturing Unit) of Anand,
India. The cheese whey had the following character-
istics (w/v): lactose, 4.5-5%; protein, 0.6-0.7%; salt
content (NaC1), 4-6%; total solids, 11-12%; volatile
solids, 4.5-5.0%; COD, 6-8%; varying amounts of
minerals and water soluble vitamins. The pH of the
whey varied from 4-5 to 5.5.
The following surfactants were used in our study.
Tegopren (T-3022), a speciality surfactant based on
polyether-polymethyl siloxane copolymers, was
obtained from Gold Schmidt AG, Essen, Germany.
The following surfactants were obtained from the
Central Drug House (CDH), India. Tween-80 (poly-
ethylene sorbitane mono-oleate); Triton-X-100
(iso-octylphenoxy-polyethoxy-ethanol); sodium lauryl
sulphate; Cetrimide (tetradecyl trimethyl ammonium
bromide).
Reactor systems
Laboratory scale anaerobic upflow fixed film reactors
consisted of a glass column with a void volume of
! litre, packing height 900 mm and packing volume
1.5 litre. Reactors were packed with charcoal as the
support material having an average size of
5 x 5 x 5 ram. Biofilms were allowed to develop on the
supporting materials for 60 days using effluent from
another operating whey reactor after mixing with salty
quick sand of coastal area, dug from about 60cm
below the surface, as initial inoculum. This initial
inoculum was slowly replaced by salty cheese whey
diluted with total dairy waste-water (1:2 v/v) in order to
obtain a final total solid (TS) of 3% (w/v) which was
filtered through muslin cloth to remove flocs formed
after adjusting the pH to 7.0 with lime. All reactors
were operated at 37 + I°C. Steady state conditions were
determined on the basis of constant COD of the
effluent. All reactors were operated for 60 days after
reaching the steady state condition. Surfactants were
incorporated into the feed. The feed was pumped
upwards continuously and the flow rate was adjusted
with the help of a peristaltic pump (Gilson, Miniplus 3
model). The reactor was operated at an HRT of 2 days.
This was found to be the most suitable from our earlier
experience [5].
Experiments were carried out in quadruplicate for
each surfactant and for each concentration.
Analytical methods
Gas production was measured by the displacement of
acidified saturated salt solution making due correction
for atmospheric pressure and temperature. Gas compo-
sition was analysed with a chromatogram (Sigma) fitted
with 2 m long stainless steel Porapak-R (80-100 mesh)
column at 40°C and thermal conductivity detector.
Nitrogen was used as a carrier gas at a flow rate of
40 ml/min. The temperatures of injector and detector
were kept at 125°C. Fatty acids were analysed using gas
chromatogram (Sigma) with a 10% FFAP column and
a flame ionization detector. Column temperature was
maintained at 180°C, whereas injector and detector
temperatures were kept at 250°C. Nitrogen served as
carrier gas. Identification and percentage of different
fatty acids were based on a comparison of retention
time and peak area of unknown with standard amount
of each acid [11]. Feed and effluent samples were
analysed for pH, volatile fatty acids (VFA), COD, total
solids (TS) by standard procedures [12].
Results and discussion
The present investigations were confined to studies of
the effects of various surfactants such as Tween-80,
Tegopren-3022, sodium iauryl sulphate (SLS), Triton-
X-100 and Cetrimide on anaerobic digestion of salty
cheese whey using fixed film reactors. Steady state per-
formance data are shown in Fig. 1. Increasing the con-
centration of surfactants improved biomethanation
providing maxima and minima at various parameters
(Fig. 1). Amongst the various surfactants used, sodium
lauryl sulphate showed the greatest effect with a
maximum gas production of 5.7 litre litre- ~ of digester
per day at 200 mg litre-1. The maximum enhancement
of over 70% was achieved with the addition of
200 mg litre-1 SLS and gas production declined there-
after. The gas was also higher in methane content in an
SLS dosed reactor. Maximum enhancement of
methane content from 69% in control reactor to 77%
in SLS (200 mglitre-J) dosed reactor was achieved.
The methane production, presented as litre of CH4 per
gram of COD utilized, also improved by about 65% in
SLS dosed reactors in comparison to control reactors
with no surfactant, thus improving the fuel value.
One of the parameters which provides information
about the process stability is the total volatile fatty
acids [13,14[. SLS dosed (200mglitre -~) reactors
showed the highest process stability as indicated by the
lowest values of VFA. The data on the level of total
volatile fatty acids present in the reactors stabilized
with different doses of surfactants are given in Fig. 2.
Average VFA concentrations ranged from
1.10glitre -I in the reactor with no surfactant to
0-520 g litre ~ in the SLS dosed reactor. These indicate
that volatile fatty acids are consumed at a faster rate
than in control experiments. The rate limiting step in
 Surfactants in anaerobic digestion of salty cheese whey 2(11

methane fermentation often involves the degradation
of fatty acids, and this is related to the efficiency of H2
utilization by methanogenic bacteria [15,16]. In the
present study, it is clear that SLS dosing enhances the
methane forming step of the digestion process. Thus
the addition of SLS helps to maintain a low level of
hydrogen by enhancing methane formation. Otherwise,
the reactor would be stressed by accumulation of fatty
acids, which in turn maintain low production of
propionate and other reduced products [15,17]. This is
further supported by the analysis of individual fatty
acids. Propionate and butyrate were found to be the
lowest in SLS dosed reactors, indicating a balance
between the formation of fatty acids and their con-
sumption (Fig. 2).
Process performance can also be judged by lower
COD values indicating better biodegradation. COD
values decreased with increasing dose of SLS, reaching
maximum reduction at 200 mg SLS per litre. The COD
was 3.0 g litre ~ in SLS dosed reactors (200 mg litre ~)
in comparison to 7"5 g litre ~ in controls without sur-
factant. This indicates a trend of increased COD
removal with increased dose of SLS, again showing
higher bacterial efficiency in SLS dosed reactors. This
in turn helped the biomethanation process.
The other surfactants, Tween-80, Triton-X-10(I and
Tegopren-3022, also increased gas production with
enriched methane content, indicating that surfactants
in general enhance substrate conversion efficiency. An
anionic surfactant like SLS exhibited greater effects on
anaerobic digestion. Surfactants are especially noted
for their wetting qualities and have unusual properties
of micelle formation and show catalysis of organic
reactions [18]. The addition of surfactant may lead to
the formation of favourable active sites by forming
micelles that enhance the coupling of sequential reac-
•tions for conversion of polymeric substances into
soluble substances, fatty acids and finally into gases.
Orientation of the surfactant molecules at the solid-
liquid interface could render the substances readily
wettable by enzymes produced by bacteria, and there-
fore the presence of surfactant may provide a morc

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Fig. I. Steady state profile of the anaerobic digestion of salty cheese whey in the presence of various surfactants: (a) Triton-
X-100; (b) Cetrimide; (c) sodium lauryl sulphate; (d) Tegopren-3022; (el Tween-80. Operational conditions: temperature;
37_+1°C; retention time, 2days; COD in influent, 30glitre ~. Symbols: gas production, (>; COD removal (%), E~; methane
content, ~.; methane production, A.
202 17.Patel & D. Madamwar

1.200 0t680 Iu ) ( b) 0.3t,0 0.300

1.150 0,660
0- 320 0.2 80
1.100 0" 6&0
0. 300 0.2 60
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0.280 0.2/,0
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o.9so o, suo o.2~,o 0.2o0
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Fig. 2. Steady state profile of volatile fatty acids in the anaerobic digestion of salty cheese whey in the presence of various
surfactants: (a) Triton-X-100; (b) Cetrimide; (c) sodium lauryl sulphate; (d) Tegopren-3022; (e) Tween-80. Operational condi-
tions: temperature, 37_ I°C; retention time, 2 days; COD in influent, 30 g litre -~. Symbols: total volatile fatty acids, ©; acetic
acid, D; propionic acid, zx; butyric acid, o.

favourable environment for the system. This will
improve the digester performance and the quality of
gas, giving a higher fuel value. However, the cationic
surfactant, Cetrimide, was not found to be effective for
the improvement of anaerobic digestion.
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