Wat. Res. Vol. 33, No. 11, pp.

2545±2554, 1999
# 1999 Elsevier Science Ltd. All rights reserved
Printed in Great Britain
PII: S0043-1354(98)00490-4 0043-1354/99/$ - see front matter

COMPARISON OF DIFFERENT MODELS OF SUBSTRATE

AND PRODUCT INHIBITION IN ANAEROBIC DIGESTION
MAREK MOÈSCHE and HANS-JOACHIM JOÈRDENING*
Lehrstuhl fuÈr Technologie der Kohlenhydrate, Technical University of Braunschweig, Langer Kamp 5,
38106 Brunswick, Germany

(First received February 1998; accepted in revised form November 1998)

AbstractÐIn batch-experiments (in lab-scale ¯uidized bed reactors) the inhibition of acetate- and pro-
pionate-degradation by propionate (substrate inhibition) and the inhibition of propionate degradation
by acetate (product inhibition) was studied. Various models were compared by ®tting them to the ex-
perimental data. The importance of independent variation of the acid concentrations and the pH for
the experimental design is discussed.
The substrate inhibition was best described by a model of inhibition by undissociated acid with an ad-
ditional independent pH in¯uence. The inhibition by propionic acid was only slight in the practically rel-
evant range of concentrations. However, the propionate degrading bacteria were sensitive against low pH.
The product inhibition was best described with the model of competitive inhibition. The inhibition
already occurred from an acetate/propionate-ratio of 1 upwards. A complete standstill of the propionate
degradation at high acetate/propionate-ratios, as expected because of the proximity to the thermodynamic
equilibrium, was not observed. # 1999 Elsevier Science Ltd. All rights reserved

Key wordsÐanaerobic digestion, kinetics, substrate inhibition, product inhibition, pH

NOMENCLATURE S substrate concentration (mg
lÿ1)

Ace total acetate concentration (disso- w2 chi square (see equation 14)
ciated and undissociated) (mg
lÿ1) s standard deviation (mg
lÿ1)
cmeas measured concentration (mg
lÿ1)
cmod modelled concentration (mg
lÿ1) INTRODUCTION
COD chemical oxygen demand (mg
lÿ1)
HI, HPro, HS concentration of undissociated acids High concentrations of organic acids can inhibit an-
(mg
lÿ1) aerobic digestion. Normally acetate and propionate
I inhibitor concentration (mg
lÿ1) are the predominating organic acids found in the
KC modi®ed constant of competitive in- out¯ow of methane reactors. Thus the e€ects of
hibition (mg
mgÿ1) acetate and propionate on their own degradation
KEq equilibrium constant (Pa3
mol
lÿ1) and the interactions between both acids regarding
KI constant of inhibition by organic their degradation are most important.
acid (mg
lÿ1) In pilot tests the substrate inhibition of acetate
KI,pH1 constant of inhibition by low pH degradation was very weak (MoÈsche, 1998). This is
KP conventional constant of competi- in accordance with other authors (Witty and MaÈrkl,
tive inhibition (mg
lÿ1) 1986; Aguilar et al., 1995). Propionate, on the con-
KS saturation constant (mg
lÿ1) trary, is frequently regarded as a stronger inhibitor
K0 product inhibition constant in (Andrews, 1969; Witty and MaÈrkl, 1986; Barredo
equation 11 (mg
mgÿ1) and Evison, 1991). Thus the investigation of sub-
P product concentration (mg
lÿ1) strate inhibition concentrates on the substrate inhi-
P(F) F probability distribution bition of propionate degradation and the inhibition
pKA negative logarithm of the ionization of acetate degradation by propionate, which has
constant been described with a similar kinetic model (Witty
Pro total propionate concentration and MaÈrkl, 1986).
(mg
lÿ1) The other case of interaction between both acids,
r reaction rate (g
lÿ1
dÿ1) the inhibition of propionate degradation by acetate,
is an example of product inhibition. In contrast to
the often-mentioned product inhibition by hydrogen
*Author to whom all correspondence should be addressed. comparatively few studies have been done on this
[Tel.: +49-531-3800950; fax: +49-531-3800988; e-mail: subject (Kaspar and Wuhrmann, 1978; Gorris et
a.joerdening@tu-bs.de]. al., 1989; Alonso, 1992).
2545
2546 Marek MoÈsche and Hans-Joachim JoÈrdening
In this study we compare several kinetic models
of substrate and product inhibition in order to ®nd
out which one describes the experimental data best.
For a signi®cant discrimination of the kinetic
models a careful experimental design and a statisti-
cal evaluation of the results are necessary.
KINETIC MODELS
Substrate inhibition
The substrate inhibition of microbial reactions
can be described by
dS rmax
ˆrˆ …
dt …1 ‡ KS =S †
…1 ‡ S=KI †
Considering that propionate can inhibit both acet-
ate and propionate degradation, equation 1 must be
rewritten more generally:
rmax
rˆ …
…1 ‡ KS =S †
…1 ‡ I=KI †
The substrate S is either acetate or propionate, the
inhibitor I is propionate in both cases.
Andrews (1969) proposed to consider only the
undissociated acid as inhibiting agent (equation 3).
The concentration of undissociated acid increases
with the total acid concentration as well as with
falling pH (equation 4).
rmax
rˆ …
…1 ‡ KS =HS†
…1 ‡ HI=KI †
with
S brium (Kaspar and Wuhrmann, 1978). Hoh and
HS ˆ and
1 ‡ 10…pHÿpKA †
…
I considers both the thermodynamic equilibrium and
HI ˆ
1 ‡ 10…pHÿpKA †
Although the Andrews model is widely accepted, it
has not yet been tested in comparison with other
models that include an independent pH in¯uence.
This is now done with this study. One alternative
model is a combination of an inhibition by the total
organic acid and an independent pH in¯uence
through a non-competitive inhibition by H+ and
OHÿ-ions (equation 5). The equation becomes more
graphic, if the pH±inhibition-constants KI,pH indi-
cate the pH, at which the reaction rate is reduced
to half of the maximum (equation 5a).
rˆ
…1 ‡ KS =S †
…1 ‡ I=KI †
…1 ‡ H‡ =KH‡ †
…1 ‡ OHÿ =KOHÿ †
rˆ
…1 ‡ KS =S †
…1 ‡ I=KI †
…1 ‡ 10…KI,pH1 ÿpH† †
…1 ‡ 10…pHÿKI,pH2 † †
In a third model the above mentioned independent pH in¯uence is combined with a substrate inhibition by
the undissociated organic acid:
rˆ
…1 ‡ KS =HS†
…1 ‡ HI=KI †
…1 ‡ 10…KI,pH1 ÿpH† †
…1 ‡ 10…pHÿKI,pH2 † †
Product inhibition
Most authors investigating product inhibition in
anaerobic digestion either use the model of non-
competitive inhibition (equation 7, Fukuzaki et al.,
1990a; Kus and Wiesmann, 1995) or give boundary
concentrations of acetate, above which a product
inhibition occurs (Kaspar and Wuhrmann, 1978;
Gorris et al., 1989; Alonso, 1992). Both are based
on the assumption that the decrease of the reaction
rate is only dependent on the product concen-
tration.
rmax
rˆ …7†
…1 ‡ KS =S †
…1 ‡ P=KI †
1† In contrast to that in enzyme kinetics the competi-
tive inhibition model (equation 8) is used to
describe product inhibition.
rmax
rˆ …8†
1 ‡ …KS =S †
…1 ‡ P=KP †
2† equation 8 can be rewritten in such a way, that the
direct dependence of the reaction rate on the pro-
duct/substrate-ratio is made clear:
rmax
rˆ …9†
1 ‡ …KS =S † ‡ …P=S
KC †
with KP/KS=KC.
The modi®ed competitive inhibition constant KC
is equivalent to the product/substrate-ratio, at
which the reaction rate is half-maximum (when the
3† substrate limitation is neglected).
It is known that the acetogenic propionate degra-
dation occurs close to the thermodynamic equili-
Cord-Ruwisch (1996) presented a model of product
4† inhibition in anaerobic propionate degradation that
the continuous transition to the Michaelis±Menten
kinetics far from the thermodynamic equilibrium:
rmax
…1 ÿ …P=S
KEq ††
rˆ …10†
1 ‡ …KS =S † ‡ …P=S
KEq †
Since the hydrogen degradation was not rate limit-
ing under the experimental conditions (see next sec-
tion), the partial pressure of hydrogen can be
assumed to be low and not to cause an additional
product inhibition. The same applies to formate as
a possible alternative electron donator. For that
rmax
…5†
rmax
…5a†
rmax
…6†
 Substrate and product inhibition
reason equation 10 was simpli®ed in such a way, that
only the e€ect of the acetate/propionate-ratio was
considered and the e€ects of the hydrogen, formate
and bicarbonate concentrations were neglected.
rmax
…1 ÿ …Ace=Pro
K0 ††
rˆ …11†
1 ‡ …KS =S † ‡ …Ace=Pro
K0 †
Equation 11 resembles equation 9 for the competitive
inhibition except the parentheses in the numerator,
which allows the reaction rate to equal zero at a ®nite
acetate/propionate-ratio when the thermodynamical
equilibrium is reached. This ratio is given by the con-
stant K0.
Applicability to propionate degradation
All the models of substrate and product inhi-
bition relate to one-step reactions. However, also
the propionate degradation, which is a multi-step
reaction, can be described in that way, if the aceto-
genic propionate degradation, and neither the acet-
ate nor the hydrogen degradation, is rate limiting.
Since the used mixed culture was adapted to the
simultaneous degradation of acetate, propionate,
butyrate and lactate, the maximum degradation
rates of acetate and hydrogen were substantially
higher than their production rates from propionate
degradation. Thus, under the experimental con-
ditions, where propionate degradation was the only
acetogenic reaction, neither the acetate nor the
hydrogen degradation was rate limiting.
EXPERIMENTAL DESIGN
Substrate inhibition
For the three models of substrate inhibition the di€erent
dependence of the inhibition on the total acid concen-
tration and the pH is shown in contour plots (Fig. 1). All
the models display a slope of the reaction rate in the direc-
tion of higher acid concentration and lower pH. The only
di€erence is the shape of this slope. The e€ect of substrate
limitation is neglected in these plots, because the satur-
ation constants KS of the investigated mixed culture are
very low (<12 mg Ace
lÿ1; <3 mg Pro
lÿ1; MoÈsche and
JoÈrdening, 1998) and the experiments were conducted at
substantially higher concentrations.
In most of the studies concerning substrate inhibition
the acid concentration and the pH are not varied indepen-
dently of each other. Either only the acid concentration is
varied (Bolle et al., 1986) or only the pH (Kus and
Wiesmann, 1995) or both at the same time (Duarte and
Anderson, 1982). Di€erent shapes of the slope of the reac-
tion rate (Fig. 1) cannot be discriminated in that way.
Only Fukuzaki et al. (1990a,b) varied the pH and acid
concentration independently. They showed that the sub-
strate inhibition was caused by undissociated propionic
and acetic acid. However, it was not analyzed whether
there was an additional independent pH in¯uence.
In this study the simultaneous degradation of acetate
and propionate in batch-experiments was followed, in
order to investigate both the substrate inhibition of pro-
pionate degradation and the inhibition of acetate degra-
dation by propionate. In one series of batch experiments
only the pH was varied. In another series only the propio-
nate start concentration was varied. In a third series both
pH and propionate start concentration were varied.
2547
Because of this experimental design di€erent shapes of the
slope of the reaction rate, as plotted in Fig. 1, could be
discriminated.
Product inhibition
The product inhibition of propionate degradation by
acetate was also investigated by simultaneous degradation
of acetate and propionate in batch experiments. To dis-
criminate the models of non-competitive (equation 7) and
competitive inhibition (equation 9) the propionate and
acetate concentrations had to be varied independently.
This requirement was not ful®lled in those studies, which
described the product inhibition with the non-competitive
model (Fukuzaki et al., 1990a; Kus and Wiesmann, 1995).
To discriminate the models of competitive inhibition
(equation 9) and the equilibrium based model
(equation 11) the acetate/propionate-ratio had to be varied
over a wide range.
These conditions were realized in a series of experiments
with di€erent levels of propionate concentration. During
each of the experiments the acetate concentration was var-
ied in the same way, but the acetate/propionate-ratios
were di€erent.
MATERIAL AND METHODS
Microorganisms
Biomass covered granular pumice (0.1±0.4 mm diam-
eter) from a 10 m3-¯uidized bed reactor for anaerobic
puri®cation of sugar factory wastewater was used (OLR:
<30 kg mÿ3 dÿ1; HRT: >12 h; pH: 6.75±7.1; CODin:
7500 mg lÿ1; CODout: 750 mg lÿ1) (MoÈsche, 1998). The or-
ganic dry matter content was 32 kg
mÿ3. Electron micro-
scopic analysis of the bio®lm revealed mainly ®lamentous
and rod shaped bacteria resembling Methanothrix sp.
(MoÈsche, 1998).
Reactors
Fluidized bed reactors with a total volume of 0.84 l and
an inner diameter of 59 mm were used. 0.3±0.4 l of the
carrier was ®lled into the reactors. The super®cial velocity
was adjusted in that way that the ¯uidized bed expanded
by factor 1.5. The temperature was maintained at
37.020.28C by pumping warm water through a heating
jacket.
Batch experiments
Substrate inhibition. Three series, each consisting of four
single batch experiments, were conducted. In each single ex-
periment a pulse of acetic acid (start concentra-
tion1 400 mg
lÿ1) and propionic acid (start concentration
varied) was fed to the reactor.
No. 1: four experiments at di€erent pH-values (7.0,
6.6, 6.2, 5.8) and low propionate start concentrations
(100±200 mg
lÿ1).
No. 2: four experiments at pH 7.0 and di€erent pro-
pionate start concentrations (1200, 600, 1100 and
1500 mg
lÿ1).
No. 3: four experiments at decreasing pH-values (7.0,
6.6, 6.2, 5.8) and increasing propionate start concen-
trations (1200, 700, 1200 and 1700 mg
lÿ1).
In the single experiments the degradation of acetate and
propionate was followed until most of the acetate was
used up (approximately 1±1.5 h). During that time 6
samples were taken. When the propionate concentration
was above 1250 mg
lÿ1, double determinations were con-
ducted. Altogether, 84 acetate and 84 propionate concen-
trations were recorded and evaluated. The pH was kept
constant (20.05) by dosage of H3PO4.
2548 Marek MoÈsche and Hans-Joachim JoÈrdening

Fig. 1. Inhibition by organic acid and pH (lines of equal degradation rate as a function of total acid
concentration and pH) according to the three models of substrate inhibition (equations 3, 5a and 6).

Product inhibition. Three experiments with di€erent Analytical methods

start concentrations of propionate (No. 1: 150 mg
lÿ1, No. The organic acids were separated by a 300 mm anion
2: 500 mg
lÿ1, No. 3: 1400 mg
lÿ1) were conducted. First exclusion column and detected by UV absorption at
the propionate degradation was followed in the absence of 210 nm.
acetate (1maximum reaction rate). Then the consumed
amount of propionate was replaced and a pulse of Evaluation methods
2500 mg
lÿ1 acetate was added. So the acetate concen-
trations in the reactors were the same but the acetate/pro- The set of di€erential equations for the simultaneous
pionate-ratios di€ered. Since the acetate degradation degradation of acetate and propionate
subsequently was much faster than the propionate degra- dPro
dation, the acetate/propionate-ratio changed during that ˆ ÿrPro …Ace, Pro† …12†
dt
phase of the experiment.
During the experiment the pH was maintained at 7.0 by dAce
dosage of H3PO4. In each experiment 14 samples were ˆ ÿrAce …Ace, Pro† ‡ rPro …Ace, Pro†
YAce=Pro …13†
analyzed for acetate and propionate concentrations, so dt
that altogether 42 acetate and 42 propionate concen- was integrated numerically (Runge±Kutta procedure). For
trations were recorded and evaluated. rAce and rPro the various model equations were inserted
 Substrate and product inhibition
(substrate inhibition experiments: equation 3, equation 5a
or equation 6, product inhibition experiments: equation 7,
equation 9 or equation 11).
Parameters, that had only negligible in¯uence on the ®t
of the models, were set to ®xed values: KS,Ace=6 mg
lÿ1,
KS,Pro=1 mg
lÿ1, KI,pH2=14.4 ÿ KI,pH1 (pH-optimum: 7.2),
YAce/Pro=0.811 mg
mgÿ1=1 mol
molÿ1.
The inhibition parameters were ®tted to the experimen-
tal data with the Nelder±Mead simplex algorithm.
Thereby the w2 as sum of normalized error squares, was
minimized in order to consider both the relative and the
absolute portion of the analytical error:
Xn   of the measured data. This primarily a€ects the cal-
ci,meas ÿ ci,mod 2
w2 ˆ …14†
iˆ1
s…cmeas †
with s(cmeas) = 0.012
cmeas+0.95 mg/l (experimentally
determined).
To ascertain, whether the di€erences between the w2 are
signi®cant, an F-test was conducted. The obtained values
of the F probability distribution P(F) represent the prob-
ability that a poorer ®tted model describes the data better
than the best ®tted model.
In Figs 2 and 3 the relative degradation rates are given.
These are the mean degradation rates in each single exper-
iment normalized to rmax obtained from the ®t of
equation 6 to all data.
RESULTS AND DISCUSSION
Substrate inhibition: model discrimination
The model of inhibition by undissociated acid
and pH (equation 6) describes the measured data
best: the w2 (normalized error sum of squares) is the
smallest (Table 1). The model of inhibition by
undissociated acid without independent pH in¯u-
ence (equation 3) is second best and the model of
inhibition by the total acid gives the poorest ®t
despite the consideration of an additional pH-in¯u-
ence (equation 5a).
All the concentration curves of the 12 single ex-
periments cannot be plotted here, to illustrate the
di€erent ®ts of the models. However, in Figs 2 and
3 the two best ®tted models (inhibition by undisso-
ciated acid with and without independent pH-in¯u-
ence) are compared with the mean degradation
rates in each single experiment.
Regarding the inhibition of acetate degradation
(Fig. 2) the predictions of both models look similar
over a wide range. This is because of the little ad-
ditional pH-in¯uence in equation 6 (KI,pH=4.7).
Nevertheless it can be seen that the decrease of the
pH to 5.8 at low propionate concentration actually
lowered the degradation rate down to 89%, whereas
the increase of the propionate concentration at pH
7.0 did not a€ect the degradation rate signi®cantly.
According to the model without additional pH-in-
¯uence (equation 3) both should have had approxi-
mately the same e€ect.
Regarding the inhibition of propionate degra-
dation (Fig. 3) the di€erences between both models
are more distinct because of the stronger additional
pH-in¯uence in equation 6 (KI,pH=5.9). Again the
increase of the propionate concentration at pH 7.0
2549
did not a€ect the degradation rate. However, the
inhibition by the decrease of the pH was rather
strong.
In the case of propionate degradation (Fig. 3)
there are noticeable deviations between the predic-
tions of the best ®tted model and the degradation
rates determined from the single experiments. This
is caused by the worse signal/noise ratio. In com-
parison with acetate, the degradation rate is low
and the higher concentrations cause a higher scatter
culation of the propionate degradation rates from
the single experiments. In the parameter ®t of the
various models the higher number of data compen-
sates this scatter.
The decision, whether the postulated additional
pH-in¯uence is really signi®cant, has to be made on
a statistical basis considering the number of
measured data as well as the goodness of ®t and the
number of parameters of the various models. This
was done with the F-test. It is shown that both of
the inferior models are very unlikely to describe the
experimental data better than equation 6.
The result, that the model of inhibition by undis-
sociated acid with an additional independent pH-in-
¯uence is the most adequate, is trivial as well as
revolutionary: on the one hand the additional e€ect
by pH can simply be explained with the pH optima
of the involved microorganisms. On the other hand
this e€ect has been neglected up to the present in
most of the mathematical models of anaerobic
digestion.
Quanti®cation of substrate inhibition
The inhibition constants for the model of inhi-
bition by undissociated acid and pH are given in
Table 1. According to that model the inhibition of
acetate degradation is very slight in the practically
relevant range (ProR 800 mg
lÿ1, pHr 6.5; see
Fig. 2(B)), whereas a certain inhibition of propio-
nate degradation can be expected at low pH (R6.6;
see Fig. 3(B)).
The calculated inhibition parameters, except
KI,pH (Pro), are outside the studied concentration-
and pH-range. A more exact determination of the
parameters would have been possible, if even more
drastic conditions would have been applied with
accordingly stronger resulting inhibition. However,
on the other hand it was the main aim to investi-
gate the inhibition in, or at least near, the practi-
cally relevant range.
The inhibition constants KI for inhibition by pro-
pionic acid are high in comparison with some litera-
ture data (Table 2). These cited values were
obtained with suspended microorganisms.
Immobilized biomass, as used in this study, is often
considered to be more resistant against inhibition.
This was attributed either to mass transport limi-
tation (Duarte and Anderson, 1982) or di€erent mi-
crobial composition (Morvai and Hollo, 1992). The
2550 Marek MoÈsche and Hans-Joachim JoÈrdening

Fig. 2. Inhibition of acetate degradation by propionate according to the models of inhibition by undis-
sociated acid (A) and with an additional pH-in¯uence (B), both ®tted to the experimental data. The ex-
perimental conditions of the single experiments (pH, start concentration and ®nal concentration of
propionate) are shown by the fat bars (q). The percentages indicate the relative degradation rates in
the single experiments.

former explanation does not apply to the biomass
used in this study: it was theoretically assessed that
the di€erence of the inner and outer concentrations
of the bio®lm were below 40 mg
lÿ1 total acid
(MoÈsche, 1998). This was con®rmed by very low
saturation constants (MoÈsche and JoÈrdening, 1998).
Regarding the microbial composition the bacteria
were predominantly rod-shaped or ®lamentous like
Methanothrix sp. This is in accordance with the ob-
servations of Morvai and Hollo (1992), who found
Methanothrix sp. in granular sludges, which were
less sensitive to substrate inhibition. On the other
hand the observed resistance to low pH has rather
been attributed to some species of Methanosarcina
but not to Methanothrix sp. (Holt et al., 1994).
The propionate degrading bacteria of the studied
mixed culture were more sensitive to low pH
(KI,pH=5.9). Heyes and Hall (1983) enriched two
subpopulations of propionate degrading bacteria:
one quickly growing acid tolerant group with high
KS and one slowly growing acid sensitive group
with low KS. The growth rate and KS of the propio-
nate degrading bacteria of our mixed culture rather
correspond to the second acid sensitive group.
Considering the cited literature, it can be said
that there are various kinds of bacteria, sensitive as
 Substrate and product inhibition 2551

Fig. 3. Substrate inhibition of propionate degradation according to the models of inhibition by undisso-
ciated acid (A) and with an additional pH-in¯uence (B), both ®tted to the experimental data. The ex-
perimental conditions of the single experiments (pH, start concentration and ®nal concentration of
propionate) are shown by the fat bars (q). The percentages indicate the degradation rates in the single
experiments.

well as insensitive to inhibition by organic acid and determined experimentally, taking into account
low pH. The kinetic data for each mixed culture the independent in¯uences of undissociated acid
are dicult to predict, so that they have to be and pH.

Table 1. Substrate inhibition models: Inhibition constants, w2 (normalized error sum of squares) and F-test (probability)

Inhibition by equation Undissociated acid (equation 3) pH and total acid (equation 5a) pH and undissociated acid (equation 6)

KI (Ace) 185 mg HPro
lÿ1 6600 mg Pro
lÿ1 290 mg HPro
lÿ1

KI (Pro) 27 mg HPro
lÿ1 1020 mg Pro
lÿ1 260 mg HPro
lÿ1
KI,pH1 (Ace) ÿ 5.5 4.7
KI,pH1 (Pro) ÿ 6.1 5.9

w2 505 621 299

P(F) 0.0014 0.00001 ÿ

(Ace): e€ect on the acetate degradation; (Pro): e€ect on the propionate degradation.
2552 Marek MoÈsche and Hans-Joachim JoÈrdening

Table 2. Inhibition by undissociated propionic acid. Comparison 52%; No. 3: 77% of the initial degradation rate).
of literature data and the results of this investigation
This leads to the conclusion that not the acetate
Target of inhibition KI (mg Pro
lÿ1) Refs. concentration, but the acetate/propionate-ratio is
relevant for the inhibition. This corresponds with
Acetate degradation >10000 Aguilar et al. (1995)
Acetate degradation 730 Jarrell et al. (1987)
the model of competitive inhibition. This model
Propionate degradation 59 Fukuzaki et al. (1990a) also describes the subsequent acceleration of pro-
Methane formation 72 Witty and MaÈrkl (1986) pionate degradation because of the decreasing acet-
Total process 40 Andrews (1969)
Acetate degradation 290 this investigation ate concentration more correctly.
Propionate degradation 260 this investigation The equilibrium based model also postulates an
inhibition dependent on the acetate/propionate-
Product inhibition: model discrimination ratio. However, the higher w2 (Table 3) and the sys-
The model of competitive inhibition gives the tematic course of the residuals (Fig. 5) show that
best description of the measured data (least w2 see this model gives an inferior description of the
Table 3). The model of non-competitive inhibition, measured data compared with the model of com-
although it is the most frequently used, gives the petitive inhibition. The signi®cance of this statement
poorest ®t. The reason can be seen from the con- is con®rmed by the F-test (Table 3).
centration curves in the three experiments (Fig. 4). At ®rst sight this result is surprising, because the
The propionate degradation is slowed down due to thermodynamical basis of the equilibrium based
the acetate pulse. The slowdown is less, the higher model cannot be wrong. However, the reason is
the propionate concentration is (No. 1: 22%; No. 2: that only the acetate/propionate-ratio was con-

Table 3. Product inhibition models: inhibition constants, w2 (normalized error sum of squares) and F-test (probability)

Inhibition equation Non-competitive (equation 7) Competitive (equation 9) Equilibrium-based (equation 11)

Inhibition constants KI=900 mg
lÿ1 KC=4.7 mg
mgÿ1 K0=28 mg
mgÿ1

w2 3200 170 400
P(F) 10ÿ23 ÿ 0.0006

Fig. 4. Acetate (above) and propionate (below) concentrations in the product inhibition experiments.
Measured concentrations (symbols) and model calculations (lines). (No. 1±No. 3: various experiments
with di€erent initial propionate concentration).
 Substrate and product inhibition
sidered and a constant hydrogen partial pressure
was assumed. However, this assumption is wrong:
the more the propionate degradation is slowed
down by a high acetate/propionate-ratio, the less
hydrogen is formed. Thus the hydrogen partial
pressure decreases and the thermodynamic equili-
brium is in¯uenced positively, so that the inhibition
due to the acetate/propionate-ratio is partially com-
pensated. A complete inhibition cannot be reached,
since the hydrogen partial pressure would then
approach zero.
So an inhibition, qualitatively similar to the com-
petitive inhibition, would result from the equili-
brium correlated model if the hydrogen partial
pressure was considered. However, model calcu-
lations with simulated hydrogen partial pressure
gave a too slight inhibition at high acetate/propio-
nate-ratios (MoÈsche, 1998). To clear up the e€ect of
the thermodynamic equilibrium on the product inhi-
bition kinetics de®nitely further experiments includ-
ing the measurement of very low hydrogen partial
pressures and formate concentrations are necessary.
Quanti®cation of product inhibition
The product inhibition by acetate was well
described with a modi®ed competitive inhibition
constant (KC) of 4.7 mg
mgÿ1 (Table 3). That
means that the propionate degradation rate was
Fig. 5. Deviation between modelled and measured propionate concentrations in the product inhibition
experiments. (No. 1±No. 3: various experiments with di€erent initial propionate concentration).
2553
half maximum at an acetate/propionate-ratio of
4.7 mg
mgÿ1 and 82% of the maximum at an acet-
ate/propionate-ratio of 1 mg
mgÿ1. A ratio of
1 mg
mgÿ1 or more is quite common in many an-
aerobic digesters. Denac (1986) reported an even
stronger competitive inhibition of propionate degra-
dation by acetate (KC=0.5±1.4 mg
mgÿ1). These
results show, that the product inhibition of propio-
nate degradation by acetate is relevant even in nor-
mal operation of digesters and should neither be
neglected in mathematical models of anaerobic
digestion nor in the interpretation of experimental
data.
The practical relevance of product inhibition by
acetate is not a€ected by the still open question of
the exact in¯uence of hydrogen via the thermodyn-
amic equilibrium: the hydrogen partial pressure was
supposed to be very low in the experiments.
Therefore, the product inhibition would even be
stronger under practical conditions, when the degra-
dation of further hydrogen sources (e.g. butyrate)
causes an elevated hydrogen partial pressure.
CONCLUSIONS
. The Andrews model of inhibition by the undisso-
ciated acid was not sucient to describe the sub-
2554 Marek MoÈsche and Hans-Joachim JoÈrdening
strate inhibition at various pH values. An inde-
pendent pH-in¯uence had to be added to that
model.
. The acetate degradation was only slightly inhib-
ited in the practically relevant range of propio-
nate concentrations and pH values. However, the
propionate degrading bacteria were sensitive
against low pH.
. The product inhibition was best described with
the model of competitive inhibition. The inhi-
bition already occurred from an acetate/propio-
nate-ratio of 1 upwards, which is in the range of
normal operating conditions of digesters.
. A complete standstill of the propionate degra-
dation at high acetate/propionate-ratios, as
expected because of the proximity to the thermo-
dynamic equilibrium, was not observed. The
e€ect of the thermodynamic equilibrium on the
product inhibition kinetics should be studied
more in detail.
AcknowledgementsÐThis work was supported by the FEI
(Forschungskeis der ErnaÈhrungsindustrie e.V., Bonn), the
AIF and the Ministry of Economics, Project No. 3769.
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