Biotechnol. Prog.

2007, 23, 767−784 767

REVIEW
Foam and Its Mitigation in Fermentation Systems
Beth Junker*
Fermentation Development and Operations, Merck Research Laboratories, P.O. Box 2000, Rahway, New Jersey 07065

Key aspects of foaming and its mitigation in fermentation systems are presented. Foam properties
and behavior, conditions that affect foaming, and consequences of foaming are discussed, followed
by methods to detect and prevent foam, both without and with the use of antifoam, and their
implications. Antifoams were catalogued according to their class (e.g., polyalkylene glycols,
silicone emulsions, etc.) to facilitate recognition of antifoams possessing similar base composi-
tions. Relatively few published studies directly comparing antifoams experimentally are available,
but those reports found only partially identify clear benefits/disadvantages of any one antifoam
type. Consequently, desired characteristics, trends in antifoam application, and chemical types
of antifoams are evaluated on the basis of a thorough review of available literature reports
describing a specific antifoam’s usage. Finally, examples of specific foaming situations taken
from both the literature and from actual experience in an industrial fermentation pilot plant are
examined for their agreement with expected behavior.

Contents a little about foams but few bother to know very much” (Gaden
and Kevorkian, 1956). Today, a similar lack of collective and
Introduction 767
systematic knowledge exists. Consequently, the purpose of this
Foam and Impact of Antifoams 767 review article is to summarize past and current information
Properties and Behavior of Foam 767 concerning foam formation and its mitigation in fermentation
Conditions That Affect Foaming 768 applications.
Consequences of Foam 770
Detection of Foam 771 Foam and Impact of Antifoams
Prevention of Foam without Adding 771 Properties and Behavior of Foam. Foam is the dispersion
Antifoam of a gas in a continuous liquid phase, and thus foam dispersions
Impact of Antifoams 772 possess bulk densities closer to that of a gas rather than a liquid
Chemical Antifoams (Defoamers) 773 (Vardar-Sukan, 1992). Foam is distinctly different from tradi-
Desired Characteristics 773 tional gas holdup for which the gas-to-liquid volume ratio is
General Trends in Application 775 smaller and bubbles are more spherical (Prins and van’t Riet,
1987). Foam bubbles are located on top of rather than within
Types of Antifoams 776
the broth, as is the case for gas holdup (van’t Riet and Tramper,
Specific Foaming Situations 780 1991). A general definition of foam, applicable to bioreactors,
Literature Reports 780 determines foam to occur when gas holdup in a gas-liquid
Qualitative Pilot-Scale Foam Observations 780 dispersion is greater than 90% (Schubert et al., 1993). Other
Summary 781 authors have quantified the gas content of foam to be in the
range of 60-90% (van’t Riet and Tramper, 1991), with a gas-
to-liquid volume ratio >1and usually >3 (Prins and van’t Riet,
Introduction 1987).
Foaming is labeled a “general nuisance” in fermentation Foam generation is autocatalytic in some cases, exhibiting a
(Weng et al., 1997) because it is necessary to adequately aerate balance between the forces creating and destroying it (Vardar-
broth while keeping foam formation under control (Vardar- Sukan, 1992). Foam layers are dynamic in nature, remaining
Sukan, 1992). During industrial fermentations, foam control is the same size when the number of bubbles bursting equals the
perceived as an “empirical art” (Vardar-Sukan, 1992). Very few number of bubbles arriving in the foam layer (Lee et al., 1993).
submerged fermentation processes are performed without the Two main types of foam are common in fermentations: (1)
use of either natural or synthetic antifoam agents (Viesturs et Froths, sea, or “bubble bath” foams are short-lived, transitory,
al., 1982). Over 50 years ago, it was stated that “everyone knows and unstable, containing a wide range of bubble sizes. Bubbles
are polyhedral and form a honeycomb structure (Prins and van’t
* To whom correspondence should be addressed. Tel: (732) 594-7010. Riet, 1987). These foams contain significant amounts of
Fax: (732) 594-7698. Email: beth_junker@merck.com. entrained liquid and thus possess low gas-to-liquid volume
10.1021/bp070032r CCC: $37.00 © 2007 American Chemical Society and American Institute of Chemical Engineers
Published on Web 06/13/2007
768
ratios. (2) Stable foams, resembling “beer head,” are uniform,
long-lasting, and rigid. Bubbles are spherical or slightly el-
lipsoidal with diameters <2-3 mm (Lee et al., 1993). These
foams are low in liquid content and thus have high gas-to-liquid
volume ratios (Gaden and Kevorkian, 1956).
Foam is stable if gas bubbles remain separated by thin liquid
walls and do not coalesce (Gaden and Kevorkian, 1956).
Drainage, the runoff of liquid between bubbles in foam, is
dependent on the liquid viscosity and density (Gaden and
Kevorkian, 1956). Bioprocess foams tend to be non-coalescing
when the presence of surface-active agents stabilizes foams as
they form (Vardar-Sukan, 1992); in other cases foams are
unstable or metastable (Berovič, 1992).
Foam stability is determined by the number of lamellae and
the angles between them; foam is stable if three lamellae are
present at an angle of 120 degrees (Ghildyal et al., 1988). Foam
stability also is favored if the surface tension of the gas-liquid
system is less that of the pure solvent of solution (i.e., the liquid
phase) (Ghildyal et al., 1988). The surface tension of pure water
is approximately 72 dyn/cm (Hall, 1971), whereas the surface
tensions of most fermentation media range between 60 and 65
dyn/cm (Viesturs et al., 1982). The surface tension of water (or
media) is further lowered by natural surfactants (such as proteins,
lipoproteins, polypeptides, and fatty acids) to about 45-50 dyn/
cm; addition of surfactants can lower surface tensions substan-
tially to values as low as 28 dyn/cm (Evans and Hall, 1971;
Hall, 1971). Foams also can be stabilized by these surfactants
(Jenkins et al., 1993).
Conditions That Affect Foaming. Some amount of foaming
during fermentation is acceptable, but excessive foaming
requires some type of control action (van der Pol et al., 1983;
Prins and Van’t Riet, 1987). Conditions that affect the degree
of foaming during fermentation include gas introduction (i.e.,
aeration), medium composition, cell growth, metabolite forma-
tion, surface-active substance formation, and indirectly, vessel
geometry (Taticek et al., 1991; Vardar-Sukan, 1992). Knowledge
of these conditions permits optimization of fermentation pro-
cesses to minimize factors that may cause foam such as oxygen
starvation and nitrogen limitation (Nobel et al., 1994b). Neural
networks have been used to predict foam behavior and recom-
mend specific action (i.e., add/not add antifoam, adjust process
conditions) (Brown et al., 2001).
Quantification. Foaminess is a characteristic of the solution
having units of time, related to the mean residence time of gas
in foam (Lee et al., 1993). Foaminess is defined as the
equilibrium volume of foam/volumetric flowrate of gas or the
height of foam/superficial velocity for cylindrical tanks (Bum-
bullis et al., 1979; Lee et al., 1993). Quantification of foam
volume and sometimes even foam height is difficult at the large
scale (Bumbullis et al., 1979). Foaminess is a measure of
foaming capacity, independent of equipment geometry and
measurement techniques but dependent on media ingredients,
their relative concentrations, and various physical factors
(Ghildyal et al., 1988). Raising gas flowrates to increase foaming
for solutions with low foaminess is hampered by difficulty
detecting the foam/liquid interface when high air flowrates are
present (Edwards et al., 1982). Other relevant measures to
characterize foaming include the liquid volume held in the foam
(foam volume/liquid volume before foaming), volumetric foam
overflow rate, and foam volume decrease over time (Wong-
samuth and Doran, 1994; Abdullah et al., 2000).
Fermentation Phase. The formation of foam differs depend-
ing on the fermentation phase. The two different foam phases
are (1) early in the fermentation, for example, owing to proteins
Biotechnol. Prog., 2007, Vol. 23, No. 4
present in complex media or when protein precipitated during
sterilization is utilized, and specifically caused by some property
of the fermentation medium, or (2) later in the fermentation,
for example, owing to protein release from cell autolysis or some
other property of the culture itself (Hastings, 1954; Schügerl,
1985; Stanbury and Whitaker, 1984; König et al., 1979; Su,
1995). This behavior parallels that found with activated sludge
foam, namely, a white, frothy foam during start-up (first 3-4
days), followed by a sticky viscous foam later on when the
culture becomes nutrient-limited (Jenkins et al., 1993). It also
parallels the difference between frothy foam in wine fermenta-
tions owing to high rates of CO2 evolution and stable, strong
foam in beer (Edwards et al., 1982). Foaming later in the
fermentation process has been found easier to control using
antifoams (Hastings, 1954).
There are five distinctive patterns of foam behavior during
fermentation (Stanbury and Whitaker, 1984 from Hall et al.,
1973): the foam (1) remains at a constant level throughout the
fermentation, (2) undergoes a steady fall during the initial
fermentation phase and then remains constant, (3) falls slightly
during the initial phase and then rises, (4) demonstrates low
initial foaming and then rises, or (5) exhibits a combination of
two or more of the above behaviors.
The surface-active agents (e.g., proteins) that cause foam
emerge from media components, extracellular enzymes, or cell
lysis (van’t Riet and Tramper, 1991). The presence of proteins
causes foaminess at concentrations from 1 mg/L up to 1-10
g/L, after which further concentration increases decrease foami-
ness since proteins coagulate (van’t Riet and Tramper, 1991).
For example, for Penicillium cultivations in the absence of
antifoam, foaminess has been found to increase, pass through a
maximum, and then decrease (König et al., 1979).
Medium Composition and the Presence of Cells, Particles,
and Surfactants. Many aspects of the initial medium composi-
tion affect foam formation, including concentrations of salts,
proteins, and sugars, as well as the presence of alcohols (Vardar-
Sukan, 1992). For example, for soybean-based media formula-
tions, the acid-hydrolyzed product generated more foam, owing
to its higher oil content, than the microbe-hydrolyzed product
(Hall, 1971). Increases in glucose up to 24 wt % in medium
containing 1-3% soybean meal increased foam stability,
possibly by raising medium viscosity (Hall, 1971). Thus, the
selection of medium components, including their method of
manufacture and their concentration, can impact foaming. In
fact, the protein content of growth medium has been cited as
the most vulnerable factor for foaming (Vidyarthi et al., 2000).
Salt-forming substances generally increase foaming and
surface viscosity as well as decrease surface tension (Ghildyal
et al., 1988). Although solution foaminess increases with
increasing salt concentration, foam stability diminishes, most
likely owing to changes in aqueous protein stability due to salt
concentration (Vardar-Sukan, 1992; Kotsaridu et al., 1983a;
Bumbullis et al., 1979). Specifically, protein solubility increases
at lower salt concentrations (“salting in” effect), which in turn
increases foam stability. Protein solubility decreases at higher
salt concentrations, raising apparent protein concentrations in
the foam layer, which in turn increases foaminess (Prins and
van’t Riet, 1987). Interestingly, low molecular weight (3000-
10,000 Da) proteins and glycoproteins promoted foam more than
higher molecular weight ones (>50,000 Da) (Hall, 1971).
Short chain alcohols (such as methanol, ethanol, or propanol)
increase foaminess of protein/water solutions with a maximum
effect at 1-2% v/v (van’t Riet and Tramper, 1991; Vardar-
Sukan, 1992). Alcohol additives also increase effective protein
Biotechnol. Prog., 2007, Vol. 23, No. 4 769

Table 1. Summary of the Influence of Fermentation Operating Conditions on Foam Formation.

element general influence mechanism comments reference
gas flow increases with higher greater amount of bubbles often foam maximum Vardar-Sukan, 1992;
rate flow rates erupting from surface exists and then decreases Van’t Riet and Tramper, 1991
superficial increases with higher greater speed of bubbles if vvm held constant with Pandit, 1989
velocity velocities erupting from surface scale up, superficial velocity
increases
sparger increases with smaller smaller bubbles form sintered spargers may Chisti, 1993
orifice orifices smaller foam cell produce high amounts
size structures which of foam
collapse more slowly
agitation increases with higher gas entrainment; cell depending on relative Hoeks et al., 1997,2003;
rate rates lysis owing to high broth/impeller geometry, Pandit, 1989
shear environment possibly can decrease foam
by providing mechanical
disturbance
viscosity increases as viscosity decreases film drainage system-dependent behavior Prins and Van’t Riet, 1987
rises above initial which increases foam
water-like values
temper- decreases with higher decreases viscosity which foaming can increase when Prins and Van’t Riet, 1987;
ature temperature increases liquid film broth cooled awaiting Gaden and Kevorkian, 1956
drainage and reduces harvest
foam
broth increases when near proteins least soluble foaming can increase Vardar-Sukan, 1992;
pH protein isoelectric near pI protein denaturation van’t Riet and Tramper, 1991
point (pI)
sterili- increases with longer Maillard reaction products foam may decrease after Kotsaridu et al., 1983b;
zation sterilization hold formed from sterilizing aeration applied for a Vardar-Sukan, 1992;
times, higher temper- nitrogen sources and duration since Maillard Schügerl, 1985
ature, higher pre-steri- reducing sugars together reactions partially
lization pH reversible

concentrations in the foam layer, causing higher foaminess but
reducing stability in some instances (Bumbullis and Schügerl,
1979; Wilde et al., 2003). In addition, carbohydrates and R-keto
acids enhance protein-containing foam stability, presumably by
affecting gas-liquid interfacial activities (Noble et al., 1994b).
Components of distillery fermentation broths, which include
sugars, electrolytes, proteins, and acids, concentrate at the gas-
liquid interface causing elastic foam that entraps fermentation
gases (Romualdo et al., 2002). Lipids, consisting of longer chain
fatty acids (g12 carbons), reduce foam in beer, competing with
proteins to weaken liquid films and destabilize bubbles (Wilde
et al., 2003).
The presence of cells, as well as secreted compounds, can
influence foam formation and stability. The presence of finely
divided, insoluble particles can assist in foam stabilization
(Schügerl, 1985) by concentrating their presence at gas-liquid
interfaces (Gaden and Kevorkian, 1956). Specifically, aqueous
solutions of colloidal materials adsorb at surfaces (i.e., inter-
faces) and add mechanical strength to the films formed (Gaden
and Kevorkian, 1956). In fermentation applications, foams can
be stabilized by hydrophobic particles, such as Nocardia cell
solids, since this culture has long-chain, hydrophobic mycolic
acids on its cell surface (Jenkins et al., 1993). Generally, thick
cell suspensions exhibit less foaming than dilute broths perhaps
as a result of higher viscosity, but at very high viscosities foam
can reappear (van’t Riet and Tramper, 1991). In some thick
mycelial fermentations, there is only a gradual increase in the
gas proportion in liquid from the bottom up to the top of the
liquid height, making it harder to quantify foam levels. However,
the effect of cells is hard to distinguish since cells are always
present together with some amount of protein (van’t Riet and
Tramper, 1991).
Foams in fermentations are likely derived from a variety of
excreted products or cell lysis products and not solely from
extracellular proteins (Vardar-Sukan, 1992; Noble et al., 1994ab).
For example, stable foams are formed when the yeast Moniliella
became nitrogen-limited and are stabilized by secreted polysac-
charides. This foam could not be suppressed using antifoam or
mechanically destroyed using foam breakers (Burschapers et
al., 2002). Removal of cells from medium due to foam formation
can cause autolysis, which releases microbial proteins that
enhance foam stability (Stanbury and Whitaker, 1984). Micro-
organisms trapped in foam experience oxygen and nutrient
limitations (Pandit, 1989; Vogel, 1983), causing changes in
microbial metabolism, protein denaturation, and/or microbial
lysis (Vardar-Sukan, 1992). Proteins and microorganisms also
are concentrated by froth flotation (Vardar-Sukan, 1992).
Cells caught in stable foams form crusts or meringues, often
adhering to walls (i.e., forming a ring) and cause sampling non-
uniformity (Abdullah et al., 2000). This problem is especially
prevalent in plant (Wongsamuth and Doran, 1994) and certain
fungal cultures. Many plant cells secrete polysaccharides and/
or proteins, causing cultures to become sticky, entrapping
bubbles in foam that then entraps cells forming a crust (Taticek
et al., 1991).
The onset of substantial cell growth reduces foaming,
suggesting metabolism modifies the composition of surface-
active agents (Bungay et al., 1960); thus some metabolites
directly and others indirectly act as antifoams (Soifer et al.,
1974). Foaming decreases in extent and duration with greater
inoculum age during production fermentations of Actinomyces
streptomycini owing to increases in antifoaming metabolite
concentrations (Soifer et al., 1974). In fact, the bacteria
Xenorhabdus has been demonstrated to produce antifoam during
its normal metabolism (Jewell and Dunphy, 1996). The creation
(via recombination by crossing strains) and/or selection (via
mutation) of a non-foaming strain of a commercial organism
helped control foaming late in fermentation (Stanbury and
Whitaker, 1984). The use of such foam-negative mutants
770
(Ishizuka et al., 1989) avoids changes in fermenter operating
conditions and reduces antifoam additions.
Fermentation Operating Conditions. Fermentation operating
conditions strongly impact on the initiation and severity of foam
formation. High air flowrates, coupled with foam-stabilizing
proteins and carbohydrates present in the broth, make fermenta-
tion processes prone to foaming and particularly challenging
applications for antifoams (Pelton, 2002). A summary of these
trends is shown in Table 1.
Foam levels generally increase in height with increasing gas
flow rate since more bubbles erupt from the liquid surface and
then are converted into foam (van’t Riet and Tramper, 1991;
Vardar-Sukan, 1992). In many cases, a maximum occurs after
which foam layers decrease in size as gas flowrates become
higher (König et al., 1979). This behavior is possibly due to a
decrease in bubble monodispersity since higher flowrates raise
coalescence rates (van’t Riet and Tramper, 1991) or an increase
in mechanical disturbances when higher flowrates disengage
from the broth (Prins and van’t Riet, 1987).
The stable foam height generally increases directly with
higher gas velocities and greater liquid depth above the sparger
(Pandit, 1989). Gas superficial velocities themselves increase
upon scale-up if specific aeration rates (i.e., volume air per
volume broth per unit time) remain constant. Consequently,
superficial velocities relevant to the large scale (e.g., 600 L)
should be examined during smaller scale (e.g., 20 L) bioreactor
process development work (Hoeks et al., 1997).
Sparger orifice designs impact foaming by affecting bubble
size. For hybridoma cells cultivated in serum-containing media,
porous metal spargers (180-200 µm, 0.00018-0.0002 m)
produce foams with bubble sizes of about 0.002-0.003 m. These
foams are challenging to control because they are densely
packed. In contrast, foams produced by ring spargers, with holes
<0.001 m that emit bubbles of about 0.01-0.02 m, are easier
to control because the larger bubbles formed large foam cell
structures and most bubbles collapsed quickly (Chisti, 1993).
Larger holes in sparger rings also can reduce foaming in plant
cultures (Taticek et al., 1991).
Agitation often increases foam by increasing air entrapment
and cell lysis. As impeller speed increases, foam cell size
decreases and becomes more stable, which in turn increases the
rate of foam buildup (Pandit, 1989). Once foam has formed,
however, increased agitation sometimes reduces foam height
owing to mechanical disturbance. However, the effectiveness
of this measure depends highly on impeller position relative to
broth level. There is additional anecdotal evidence for animal
cell cultures that downward pumping hydrofoil impellers reduce
foam formation relative to other impeller designs such as
Rushton impellers.
Foam level and persistence decreases as temperature increases
potentially as a result of decreased broth viscosity (Vardar-
Sukan, 1992; Prins and van’t Riet, 1987), increased drainage
of liquid films (van’t Riet and Tramper, 1991; Gaden and
Kevorkian, 1956), and lower gas pressure within bubbles (Gaden
and Kevorkian, 1956). These observations possibly explain
foaming increases when broth is cooled while awaiting harvest.
Other authors found that temperature increases decrease foam
stability but enhance foaminess (Ghildyal et al., 1988) since
protein denaturation increases (Vardar-Sukan, 1992).
Foam formation is affected by the liquid (broth) properties,
specifically viscosity, surface tension, and ionic strength (Var-
dar-Sukan, 1992). Foam increases when viscosity rises from 1
mPa-S up to 10-100 mPa-S; beyond 10-100 mPa-S foaming
decreases, but at very high viscosities foaming starts again (Prins
Biotechnol. Prog., 2007, Vol. 23, No. 4
and van’t Riet, 1987). Liquid-phase surface tension directly
affects stable foam height (Pandit, 1989). The liquid film
thickness forming the foam bubble boundary is proportional to
σ0.57 (Pandit, 1989). Ionic strength primarily affects foaming
by altering protein solubility as discussed previously.
Broth pH affects the action of antifoam agents (Vardar-Sukan,
1992; van’t Riet and Tramper, 1991) because it affects foams
produced by colloidal agents such as proteins (Gaden and
Kevorkian, 1956). If the pH is near the protein pI (isoelectric
point) where proteins are least soluble (Vardar-Sukan, 1992),
foam generally reaches its maximum extent (van’t Riet and
Tramper, 1991). In one example, broth pH decreases from 6 to
4 increased foaming and foam stability about 10-fold for a
medium containing 4-5% soybean meal (Hall, 1971). In a
second example, foam volume increased 4-fold and foam
stability increased 6-fold for shake flask plant cell cultures of
Atropa when broth pH was lowered from 7 to 6 (Wongsamuth
and Doran, 1994).
Sterilization increases the already high foaming capacity of
complex nutrient media considerably (Vardar-Sukan, 1992;
Schügerl, 1985). Heat causes nitrogen sources to become
hydrolyzed or partially degraded, leading to Maillard reactions
between reducing sugars and amino acids/proteins/peptides;
these Maillard reaction products enhance foam formation
(Vardar-Sukan, 1992). Maillard reaction products increase with
higher sterilization temperatures, longer sterilization times, and
higher presterilization pH values (Kotsaridu et al., 1983b).
Overall Maillard reaction products can increase foaminess by a
factor of 2100 during sterilization of potato-protein liquor and
glucose medium (Ghildyal et al., 1988). During sterilization
foam-stabilizing components form in soybean flour medium
based on Maillard reaction product formation (Vardar-Sukan,
1992). Since the Maillard reaction is partially reversible, a
reduction of foaminess is observed when sterilized medium is
“aged” by aeration with sterile air (Vardar-Sukan, 1992) or
stored at low temperatures (Kotsaridu et al., 1983b). Other
specific examples of these effects have been reported: (1) The
foam stability of a medium containing 2% soybean meal, 4%
glucose, and 0.5% CaCO3 increases 5-fold during a 90 min
sterilization at 125 °C (Hall, 1971). (2) The foaming coefficient
of molasses increases 2-fold for a sterilization temperature
increase from 110 to 130 °C (Berovič, 1992).
Consequences of Foam. Sterilization. Foam during steriliza-
tion can cause sterility problems during the subsequent cultiva-
tion (Vardar-Sukan, 1992). Foaming during sterilization splashes
media (e.g., flour) particles on fermenter side walls that may
be sterilized inadequately. Foam probes can be enabled during
sterilization to detect foam formation.
Working Volume. Foaming enhances gas holdup (Vardar-
Sukan, 1992), which expands broth volume (Berovič, 1992),
often requiring reductions in the working (i.e., operating) volume
of the fermenter below the maximum level to ensure sufficient
headspace (Edwards et al., 1982; Brown et al., 2001). These
effective liquid volume increases lead to broth (e.g., substrate,
biomass) loss through air entrainment, creating several sterility
as well as containment problems (Bryant, 1970; Vardar-Sukan,
1992; Berovič, 1992). Wetting of off-gas hydrophobic vent
filters (Duitschaever et al., 1988) causes plugging and back-
pressure buildup and then potential release of pressure relief
devices (Koller, 2004). Cells that become entrapped in foam
via flotation (Bryant, 1970) or deposited on upper parts of the
fermenter undergo autolysis, which releases surface-active
agents that catalyze further foam formation (Ghildyal et al.,
1988; Vardar-Sukan, 1992). Productivity also is reduced from
Biotechnol. Prog., 2007, Vol. 23, No. 4
foam-related depositions of substrate and/or product on non-
wetted parts of the vessel and piping (Brown et al., 2001; Pandit,
1989).
Mass Transfer. Mass transfer is enhanced when sur-
face-active compounds improve bubble stability by reducing
coalescence and increasing the surface area available for mass
transfer (Noble et al., 1994b). However, two mechanisms exist
for antifoams to reduce mass transfer: (1) creation of larger
bubbles and decreased gas holdup due to antifoam-induced
bubble coalescence, and (2) creation of a spread film by the
defoamer on bubble surfaces, reducing oxygen diffusion rates
(Pelton, 2002). In the latter mechanism, mass transfer is slowed
due to antifoam accumulation at the gas-liquid interface causing
an additional gas-liquid interfacial resistance and decreased
gas diffusion (Vardar-Sukan, 1992). Mass transfer coefficients
are relatively independent of DO (i.e., concentration driving
force) without antifoam present; with antifoam present these
mass transfer coefficients become linearly dependent on DO,
possibly due to the oxygen diffusion coefficient through the
antifoam layer becoming concentration-dependent, an effect
noted for polymers (Bull and Kempe, 1971).
Foam not only causes changes in air bubble size but also in
composition, altering dissolved gas profiles due to heterogeneous
gas dispersion (Vardar-Sukan, 1992). Foam increases bubble
residence time in broth, resulting in bubbles becoming oxygen-
depleted (Stanbury et al., 1995) and accumulating carbon
dioxide, thus delaying its removal (Berovič, 1992; Koch et al.,
1995). Interestingly during early hybridoma animal cell cultiva-
tions at the 1 L scale, stable foams are observed to increase
surface aeration by 90% (Ju and Armiger, 1990), but this effect
is considered primarily an indirect result of reduced survival of
cells present in foams.
Power Input/Mixing. The presence of foam reduces apparent
viscosity owing to higher gas holdup, thus decreasing power
dissipation and circulation rates (Vardar-Sukan, 1992). Thus,
permitting some foaming (and avoiding antifoam addition) can
increase mass transfer substantially and reduce power consump-
tion (Berovič, 1992). In one example, when medium is changed
from a non-foaming to foaming system, a higher OTR is
achieved at same power input (Adler and Fiechter, 1986).
Instrumentation. Foam on the broth surface adversely affects
level measurements, which makes continuous cultivation dif-
ficult (Berovič, 1992). The presence of small bubbles from foam
also can interfere with electrode sensors (Vardar-Sukan, 1992;
Pandit, 1989).
Miscellaneous. In some difficult fermentations (no specific
information given by authors), foam must be strictly avoided
because certain broth components are rendered corrosive by high
oxygen levels in foam (PharmaTec, 2004).
Detection of Foam. Types of Foam Sensors. Several types
of foam sensors exist to detect the presence and, in some cases,
depth of foam. Conductive probes, based on conduction of
electrical charge (i.e., DC voltage) between the probe and vessel
by ions present in the liquid (Brown et al., 2001), are sensitive
to biofilm (Hall, 1971), which causes a permanent short circuit
due to the collection of material across the probe’s insulation
(Ghildyal et al., 1988). Capacitance probes have less biofilm
interference than conductive probes since the entire capacitance
probe is covered with insulation (Hall, 1971). Capacitance is
sensed as the foam becomes dielectric (Ghildyal et al., 1988).
Both conductance and capacitance foam sensors are able to
measure liquid (i.e., foam) level (Getchell, 1983; Brown et al.,
2001), possessing a user-adjustable detection level rather than
fixed point. Less common are admittance probes, which use
771
high-frequency signals with wide band measuring bridges and
amplifiers able to measure foam even with a biofilm present
(PharmaTec, 2004). Another alternative for foam detection is
sonar level detection based on the presence of a foam layer in
some cases interfering with sonar feedback.
Control Strategies. Several early examples of automated foam
detection and antifoam addition systems are noted in the
literature (Pfeifer and Heger, 1957; Bartholomew and Koslow,
1957), forming the basis for how antifoam addition is controlled
today. Since on-off foam detection only indicates that foam is
present (Brown et al., 2001), it is being replaced by continuous
level detection that gives the foam height and permits set point
adjustment. For sensors linked to automated antifoam addition
systems, small time lags are introduced to prevent over-dosing
and control the stop/start operation of the pump (PharmaTec,
2004). The time lags also permit adjustment of (1) the amount
of antifoam added, (2) the interval between antifoam additions
to permit mixing and for the antifoam to take effect, and (3)
the detection sensitivity threshold by requiring continuous
detection for a designated time period to avoid additions caused
by splashing (Reisman, 1988; Getchell, 1983). In one example
strategy, antifoam addition begins if the foam signal persists
for 5 s continuously, then alternately adds antifoam for 30 s
and mixes without antifoam being added for 30 s, repeating
this cycle until foam is not detected for 5 s before stopping.
For early antifoam addition on/off cycles, incorporation of a
time lag permitted mixing and reduced overall antifoam
consumption when compared to manual addition methods
(Bartholomew and Koslow, 1957; Dworschack et al., 1954).
The antifoam addition tubing bore size and pump rate can be
used to estimate the amount of antifoam added per shot
(although this value is somewhat dependent on fermenter back
pressure), or antifoam reservoirs can be weighed continuously.
Prevention of Foam without Adding Antifoam. Foam
prevention, without additional antifoam beyond what may be
present in the initial medium composition, is preferred if it is
reliably attainable with no deleterious effects on the process
(Vardar-Sukan, 1992). A brief summary of available options is
presented below.
Physical and Mechanical Methods Including Foam Break-
ers. Physical methods to prevent foam such as ultrasound,
thermal, or electrical treatment can adversely affect cells
(Vardar-Sukan, 1992) so these are not discussed further.
Mechanical methods reduce foam by subjecting it to shear
stress, but these devices can have significant extra power
requirements (Brown et al., 2001). In some instances, they can
increase cell death despite similar biomass concentrations (Vraná
and Seichert, 1988), and they may preferentially destroy larger
over smaller foam bubbles. Common mechanical foam breakers
use rotating elements such as discs, bladed wheels, or stirrers
(Solomons, 1969; PharmaTec, 2004; Yasukawa et al., 1991;
Takesono et al., 2001) or a simple bar attached to the agitator
shaft above the liquid level. They often are complicated in
design, have high running costs, and also are unreliable (Prins
and van’t Riet, 1987; Vardar-Sukan, 1992). Mechanical foam
breakers are used when the process cannot tolerate chemical
antifoams (Olivieri et al., 1993), as was the case for early animal
cell cultivation processes (Chisti, 1992). Foam breakers, used
in conjunction with antifoam addition, can reduce antifoam
addition by 33-50% (system not given) (Yamashita, 1972).
Agitation Rate. Although counterintuitive, stirring-as-foam-
disruption (SAFD) reduces foam by causing a higher liquid
velocity to be generated by an upper impeller strategically
located near the broth-foam (dispersion) interface. Axial
772
impellers are more effective at this task than Rushton impellers.
Specifically, for the volume range where this technique is
effective, the foam height is almost linearly dependent on broth
mass for constant Vs and rpm (Boon et al., 2000; Hoeks et al.,
1997,2003). The mechanism of SAFD is foam entrainment; the
gas holdup of the broth rises sharply when the impeller knocks
down foam. Thus, the aerated liquid volume relative to the upper
impeller location becomes a key parameter for effectiveness
(Boon et al., 2002). In this pilot plant facility, this technique is
effective for a fungal Glarea fermentation, as well as more
effective than P2000 antifoam addition for knocking down foam
induced by addition of Tween 80 surfactant.
Other Operating Conditions. Foam generally is reduced by
increasing back-pressure, decreasing agitation, and/or decreasing
aeration during cultivation. Reducing airflow or agitation rates
to reduce foam usually adversely affects productivity (Vardar-
Sukan, 1992). Although productivity often is less affected by
back-pressure, higher back-pressures may not always be attain-
able at production scales without limiting airflow rates. Increas-
ing back-pressure from 10-15 to 30 psig decreases foam during
production of itaconic acid by Aspergillus (Pfeifer et al., 1952).
Increasing back-pressure from 1.1 to 1.5 kg/cm2 when applying
air to end sterilization and until the batch is cooled to its
cultivation temperature also served to reduce foam in this pilot
plant facility.
Impact of Antifoams. Several negative effects of antifoams
are noted in the literature. In general, negative effects of
antifoams decrease when they are added regularly to fermenta-
tion processes in low amounts rather than in fewer additions of
higher amounts (Kovalev et al., 1982).
Concentration. Foam formation can be enhanced by anti-
foams present in too high concentrations (Vardar-Sukan, 1992).
Specifically, foam stability decreases up to a maximum surfac-
tant concentration, and then subsequent surfactant additions
actually increase foam stability (Vardar-Sukan, 1992). When
too much antifoam is added, foam is destroyed, but the liquid
becomes strongly coalescing (Prins and van’t Riet, 1987).
Mass Transfer. A tradeoff exists between minimizing foam
height and maximizing mass transfer (van’t Riet and van
Sonsbeek, 1992). Antifoams present at low concentrations
markedly decrease the volumetric mass transfer coefficient, KLa,
with the major effect being on the overall gas-liquid mass
transfer coefficient, KL, and some effect on the specific
interfacial area, a (Morão et al., 1999; Kawase and Moo-Young,
1990). Conditions that cause bubble collapse in foam also
promote coalescence of bubbles within the liquid phase, which
results in larger bubbles with lower specific interfacial surface
areas and thus lower mass transfer (van’t Riet and van Sonsbeek,
1992). Different types and quantities of antifoams can affect
dissolved gas dispersion to the culture and consequently overall
respiration rate (Elander, 1989). When gas holdup and specific
interfacial area are reduced suddenly by anti-foam addition, the
broth DO drops quickly (Schügerl, 1988; Yagi and Yoshida,
1974; Lengyel and Nyiri, 1966) but also recovers quickly.
In general, mass transfer rates can be lowered by antifoam
up to 50% (Berovič and Cimerman, 1979; Solomons, 1966;
Atkinson and Mavituna, 1983) or 60% (PharmaTec, 2004) or
75% (Phillips et al., 1960). For example, despite higher
interfacial area owing to a 13% reduction in surface tension,
0.25% of 3% Alkaterge C in lard oil causes a 50% reduction in
the gas (oxygen) absorption coefficient of filtered Penicillium
broth using complex medium (Deindoerfer and Gaden, 1955).
The amount of reduction varies depending on the fermentation
class: for yeast fermentation, 25% decrease in mass transfer;
Biotechnol. Prog., 2007, Vol. 23, No. 4
for penicillin fermentation, 50-70% decrease in mass transfer
(Chain et al., 1966). The amount also varies depending on the
antifoam class for fermentation of the yeast Torulopsis on
complex medium containing sugar beet molasses: control 47;
0.1% Dow Corning Antifoam A (Dow-Corning, silicone-based)
18.8; 0.1% Hodag K-4 (Lambent, organic-based) 20.4; 0.1%
lard oil (natural oil) 31.4, all in units of mmol/L-h (Phillips et
al., 1960). Finally, the amount varies based on antifoam quantity
present: specifically, the KLa of pure medium decreases sharply
upon addition of a small amount of KM70 (silicone emulsion)
antifoam (Shinetsu Kagaku); after about 200 ppm is added there
is no further reduction (Takahashi and Yoshida, 1979). Similarly,
mass transfer rates decrease from 0 to 0.01% antifoam with no
effect between 0.01% and 0.1% for Desmophen 3600 (polyester
polyol) added to protein solutions (Adler et al., 1980). A critical
antifoam concentration does exist above which both mass
transfer and gas holdup increase (Morão et al., 1999; Kawase
and Moo-Young, 1990). Owing to greater gas holdup and lower
circulation rates, foaming also can result in lower heat as well
as mass transfer rates (Vardar-Sukan, 1988).
When antifoam effects are measured in water or media
without cells, there are steep decreases in KLa reported, most
likely due to most of the antifoam accumulating around the gas-
liquid interface and interfering with mass transfer. In one
example using 3000 L fermenters, the mass transfer decrease
caused by antifoams is largest (70-85% decrease) in sulfite
solutions, less (50-70% decrease) in P. chrysogenum, and even
lower (25% decrease) in the yeast Torula utilis (Chain et al.,
1966). These results suggest that to truly characterize the effects
of antifoam on mass transfer, detrimental effects should be
characterized comparing antifoam-containing and antifoam-free
media containing cells (i.e., actual fermentation broth) since
antifoam distribution is affected by the presence of hydrophobic
cells.
Power Input/Mixing. Depending on the antifoam concentra-
tion, sharp increases in power draw due to degassing or a drop
in mixing power due to lowered gas entrainment is observed
(Ghildyal et al., 1988). In one instance, an increased power draw
of up to 20% for large fermenters is observed after addition of
a natural oil antifoam (Bungay et al., 1960).
Biological and Metabolic. The presence of antifoam can
change the growth rate and even morphology of cells (Noble
et al., 1994b; PharmaTec, 2004), also negatively affecting cell
density and/or product concentration (PharmaTec, 2004). Certain
antifoams are preferentially used as a carbon source by some
cultures (Ghildyal et al., 1988). The broth pH profile can be
altered by fatty acids released due to lipase action on oils which
both changes culture metabolism and fermentation progression
(Ghildyal et al., 1988; Elander, 1989). In one application, this
behavior is used to cause crude pH adjustment by relying on
the culture to produce fatty acids by metabolizing antifoam oils
(Bungay et al., 1960).
Instrumentation. In one report, antifoams are found to
accumulate on the membrane surfaces of DO probes, increasing
diffusion resistance (Chen and Wang, 1993). However, this
study is conducted in the absence of cells; in the presence of
cells, the cells themselves also are observed to cause significant
resistance. There are other informal reports of antifoam ac-
cumulation when concentrations exceed 1 mL/L on the surface
of filter probes, designed to permit cell-free sampling of
fermentation broths.
Downstream Isolation. Antifoams retain their surface-active
nature during isolation steps, so their permissible levels during
mid-cycle fermentation should be examined in conjunction with
Biotechnol. Prog., 2007, Vol. 23, No. 4 773

downstream processing requirements (Paul et al., 1981). Anti- Table 2. Selected Silicone-Based Oil (S) and Emulsion (SE)
foams cause difficulty in extraction and subsequent purification Antifoams Noted in Fermentation Literaturea
owing to the formation of difficult-to-break emulsions in past and/or current
aqueous-solvent systems (Ghildyal et al., 1988), specifically tradename vendor
impairing phase separation during whole broth extraction (Paul Silicone (100%) [Poly(methylsiloxane) in Silicone Oil]
et al., 1981). Antifoams also can co-extract with the desired Sag M-10 Dow Corning
product (Pollard et al., 2006). Antifoams accumulate on mi- Sag 471 Union Carbide
DC-A, A Dow Corning
crofiltration membranes causing membrane fouling and even Antifoam A (concentrate) Sigma
destruction (PharmaTec, 2004; Liew et al., 1997). Often Mazu DF100, DF1005 Noveon, Mazur
changing the filtration step temperature relative to the antifoam’s FD 101b Stepan
cloud point can enhance performance, however, and an in- Q7-2243 LVAb Dow Corning
C100F b Basildon
process assay to detect their removal is critical to efficiently S184 Wacker Chemie
developing these steps. Finally, antifoams interfere with the Q10-335b Dow Corning
binding step in ion exchange separations. Because of these TMA812 Toshiba
isolation interferences, extra separation steps, often specific in Antaphron NM40 UK
Biospumex FDA 165K Cognis
nature to the antifoam employed, are implemented to remove
defoamers (Ghildyal et al., 1988). Silicone Emulsions (Silicone Percentage)
KM-70, M-7W-0 (37%) Shinetsu Kogaku
Mazu DF 210, 210 S, 2105 (10%) Mazur/Basf/Noveon
Chemical Antifoams (Defoamers) 1510 (10%) Dow Corning
1520 (30%) Dow Corning
Antifoams and defoamers are fundamentally the same chemi- AF (30%) Dow Corning
cals despite some differing implications noted in the literature C (30%) Dow Corning
(Pelton, 2002). Antifoams are defined as strongly surface-active DC-B (10%) Dow Corning
substances which replace foam-forming components and lower FG-10 (10%) Dow Corning
DSP (10%) Dow Corning
surface tension of liquids (van’t Riet and Tramper, 1991; van’t RD (10%) Dow Corning
Riet and van Sonsbeek, 1992). Antifoams are dispersed by M30 (30%) b Dow Corning
stirring, and foam is destroyed by bubble coalescence, which Q7-2587 (30%) Dow Corning
decreases the available surface area for gas-liquid mass transfer Hodag FD-62 (10%) Lambent
(de Haut, 2001). Antifoams typically are added to medium or Hodag FD-82 K (30%) Lambent
A (A-5758) (30%) Sigma
broth before foaming occurs (Ghildyal et al., 1988), and B (10%) Sigma, JT Baker
sometimes they are called foam inhibitors (Weng et al., 1997). C (30%) Sigma
In contrast, defoamers compete with other surface-active agents Chemax DF-10 (10%) Rutgers
for the surface layer, but they do not support foam formation Chemax DF-30 (30%) Rutgers
SE15 (10%) Sigma
(Gaden and Kevorkian, 1956). Defoamers are self-dispersed, SE9 (10%) Wacker Chemie
and foam is destroyed by surface action (de Haut, 2001). Sag 10 (10%) Union Carbide
Typically, defoamers are used to knock down foam after it has Sag 30 (30%) Union Carbide
formed (Ghildyal et al., 1988), and sometimes they are called Silcolapse 5001 (10%) ACC/Rhodia
Silcolapse 5000 (30%) ICI/Ambersil/Rhodia
foam breakers (Weng et al., 1997). For the purpose of this
GE 60 (30%) GE
review, the term antifoam is used to cover both aspects. Roth 0865 (UK%) Carlroth.de
Antifoams act via four steps via antifoam droplets: (1) PD30 (30%)b Basildon
entering the liquid film around the foam bubble, (2) bridging RS-70426 (27%)b Rhodia
the width of the liquid film, (3) dewetting by causing the liquid Silicon-Based
film to thin around where the antifoam is present, and (4) rupture 1705-W Lion
Assaff III Rhone-Poulenc
of the liquid film (Garrett, 1993; Denkov et al., 1999; Jha et
al., 2000; Christiano and Fey, 2003). Certain antifoams contain Complex Polyhydric Alcohol/Silicone Polymer
SO-25b Sigma
insoluble particles which reduce the viscosity of foam lamellae,
a Information obtained from company websites, including MSDSs, and
causing bubbles to thin and drain in this fashion, and severely
reducing foam stability (PharmaTec, 2004). handbooks (Ash and Ash, 2000a,b) in addition to publications. Information
limited by proprietary disclosures. Silicone oil is a silicone polymer, also
Chemical antifoams are simple and economical (Vardar- known as poly(dimethylsiloxane). Simethicone consists of 95% poly(di-
Sukan, 1992). Consequently, they have proliferated and an methylsiloxane) (silicone oil) and 5% hydrophobic silica. Sigma Antifoam
abundance of choices are commercially available, both currently A/C emulsions are both 30% but use different emulsifiers. Sigma sells both
Antifoam A concentrate (100% silicone) and emulsion (30%). Some authors
and in the past. Tables 2, 3, and 4 compile examples of do not distinguish which type was used. b Antifoams offered commercially
antifoams, many of which are cited in fermentation literature. but not cited in fermentation literature at this time.
Unfortunately, often trade names do not aid in understanding
the nature of the compounds. Consequently, similar antifoams
Desired Characteristics. No single antifoam can possess all
are grouped together by “class” within Tables 2, 3, and 4 so
of the ideal, desirable characteristics (Vardar-Sukan, 1992), thus
that available antifoams options for fermentation applications
are readily accessible and assessable. Confidentiality concerns a compromise often is needed. Antifoam efficiency is directly
frequently preclude obtaining precise details about antifoam proportional to its foam suppression ability and inversely
compositions. In a few cases, little or no information is proportional to the amount consumed in the fermentation to
obtainable. For those cases where an antifoam is composed of suppress foam (Vardar-Sukan, 1988). This efficiency is deter-
components from more than one antifoam class, it is listed with mined by comparing the (1) minimum volume required and (2)
the class of its highest percentage composition. In instances maximum yield of product or absence of any microbial activity
where one antifoam class is dissolved in a carrier of a second (Duitschaever, 1988). Overall antifoams must have properties
antifoam class, the class of the solute is applied. that permit them to function as antifoams, be used with living
774 Biotechnol. Prog., 2007, Vol. 23, No. 4

Table 3. Selected Poly(alkylene glycol) (PAG)-Based Antifoams Noted in Fermentation Literature

chemical tradename vendor
poly(propylene glycol) (linear polyether of propylene oxide), PPG P2000 Dow
PPG2000 Wacker-Chemie
PPG 2025 E. Merck
PPG1025 (mw 1000) BDH
PPG-24 butyl ethera UK
Pluriol P2000a BASF
Pluracol P2000a BASF
Pluracol P2010 BASF
Emkaphyl Dow
poly(ethylene glycol) (linear polyether of ethylene oxide), PEG Pluracol E BASF
PEG Typ 300 Carl Roth
poly(alkylene glycol) (e.g., linear polyether of ethylene and KFO F119, F161a Noveon
propylene oxides; poly(propylene glycol) KFO 673 KABO/Lubrizol/Emerald
ether of butyl alcohol) UCON LB625 (mw 1500) Dow
Mazu DF60P Mazur
Mazu DF7960 Mazur
Mazu DF800 S, DF8005 Mazur
Ucolub N115, N-7 or N-7/1 Brenntag
Struktol J647, SB2121 Schill and Seilacher
SP1 Th. Goldschmidt
difunctional ethylene/propylene oxide (EO/PO) block copolymers Tetronic 901 BASF
Pluronic F68 BASF
Pluronic L 61, PL-61, PE6100 Ugine Kuhlmann
Pluriol L81, PE8100 BASF
Pluronic L122 BASF
Pleuronic Fluka
Adekanol LG-109, LG-107 Fermenta AB, Asahi Denka Kogyo, Adeka
Mazu DF204 PPG Ouvrie
polyalcohol based on EO/PO block copolymer Struktol J650a Schill and Seilacher
poly(alkylene glycol) (PAG)-based Breox FMT30 Intl. Specialty Chemicals/Cognis
polyether Tego KS911 Degussa, Goldschmidt
likely PAG-based: 99% organic defoamer, 1% silicon glycol to Antifoam 289, 286 Sigma
enhance spreadability
PAG-based: 20% branched simethylsiloxane/3.5% dispersed VP1133 Wacker-Chemie
silicone dioxide/76.5% PPG2000
PPG-based polyether dispersions Antifoam 204 Sigma
PAG-based with small amount of silicone XFO-371 Ivanhoe
PAG-based: 95% polyalkylene/5% silicone Sag 5693, 5698 Union Carbide, OSI Specialties
PAG derivative Disfoam GD Nihon Yushi/BASF
alkyl poly(alkylene glycol) ether blend Bioquest 1110/1120a Baker Hughes
oxyalkylated alkylphenolic resins Bioquest 1395a Baker Hughes
organic oil Hodag K-4 UKb
UK Troy 333 UK
SAG 4130 UK
Clerol FBA 622 Cognis/Henkel Nopco
DL-2000 UK
a Antifoams offered commercially but not cited in fermentation literature at this time. b UK ) unknown.

cells, and not to interfere with electrodes such as pH or DO
sensors (Vardar-Sukan, 1992).
Speed/LongeWity. Antifoam effectiveness is measured by the
rate at which foam collapses and the duration of its action both
initially upon addition and over the period it is present in the
broth (Byrant, 1970). Since often antifoams are added on
demand, initial instantaneous action is highly desirable (Vardar-
Sukan, 1992) to achieve fast foam-breaking (Stanbury and
Whitaker, 1984, from Hall et al., 1973; Vardar-Sukan, 1992)
or knock down (Berovič, 1992; Solomons, 1967). Considering
its projected elaspsed time in the broth, a short-acting defoamer
could be tolerated for fermentations that foam for only short
periods (Corbett, 1985). In general though, antifoams should
be long-lasting and non-metabolizable (Stanbury and Whitaker,
1984, from Hall et al., 1973; Vardar-Sukan, 1992; Berovič,
1992), as well as readily transferable to and dispersible in
fermentation broth (Stanbury and Whitaker, 1984, from Hall et
al., 1973).
Concentration/Cost. Antifoams should be low cost and
effective at low concentrations (Vardar-Sukan, 1992; Berovič,
1992; Stanbury and Whitaker, 1984, from Hall et al., 1973). In
some cases, a balance is explored between selection of a costlier
but less utilized (and usually more effective) antifoam and
selection of a cheaper but more highly utilized (and usually less
effective) antifoam (Corbett, 1985).
Solubility. Insolubility of antifoams is preferred to promote
their activity at low concentrations (Hall, 1971). Together with
low solubility, antifoams also should have a low critical micelle
concentration to reduce foam stability through surface interac-
tions (Lee and Tynan, 1988). They should be insoluble in the
foaming medium, specifically possessing some hydrophobic
character for low solubility but also having some hydrophilic
character to ensure a low interfacial tension and thus a positive
spreading coefficient (Vardar-Sukan, 1992; Sie and Schügerl,
1983). Determining where the antifoam resides in the fermenter
during a post-batch examination generates insight as to how it
works (Phillips et al., 1960) and its solubility. Specifically, for
spreading coefficients >0, air-oil-water systems are more
stable than air-water systems and oil likely exists as a thin
film at the air-water interface; when the spreading coefficient
is <0, oils exists as discrete droplets in the bulk liquid (Weng
Biotechnol. Prog., 2007, Vol. 23, No. 4 775

Table 4. Selected Antifoams, Based on Fatty Acids/Esters (FE), Polyesters (PEP), and Oils (NE) Noted in Fermentation Literature
chemical tradename vendor
alkylpolyalcoxyester Clerol FBA 3107a Cognis
polyester polyol Desmophen 3600, 3900 Bayer AG
ester and polyalcohol Struktol SB2023 Schill and Seilacher
fatty acid ester O-30 Sigma
fatty acids, paraffin oil and non-ionic emulsifier Contraspum 210 Zschimmer and Schwarz
alkoxylated fatty acid esters on vegetable base Struktol J673 Schill and Seilacher
polyalkoxyester (EO/PO block polymer/ester) Biospumex 153 K Cognis
poly(ethylene glycol)-fatty acid ester and silicone oil AF emulsion Nakarai
glycerol alkyl oleate Atlas G 5600 Atlas Europol-ICI
natural oils various (lard, lard burning, soybean, corn, various
maize, cod liver, cottonseed (Proflo),
olive, sunflower, safflower, peanut,
ground nut, grape seed, linseed,
poppyseed, caster, palm
additives in natural oils 8% Alkaterge in grapeseed oil various
3% Alkaterge in lard oil
1% PPG in lard oil
octadecanol (stearyl alcohol)
1% octadecanol in ethyl alcohol
3% octadecanol in soybean oil, lard oil,
or peanut oil
substituted oxazolidine Alkaterge C commercial solvents
66 Alkaterge commercial solvents
cetyl polyoxyethylene condensate Lubrol Wa ICI
likely PAG-based, some containing fatty acid esters Glanapon Bussetti/Jastbolaget
non-silicone Nopco (Foam Master) Cognis/Henkel
UKb Prochem #51 Lambent
Dehysan 2111 Henkel
a Antifoams offered commercially but not cited in fermentation literature at this time. b UK ) unknown.

et al., 1997). In addition, non-metabolizable synthetic antifoams
are likely to coat vessel surfaces, thus requiring use of larger
amounts.
Chemical Properties and Toxicity. Antifoams should have
low surface and interfacial tension (Vardar-Sukan, 1992) and
should be non-volatile (Vardar-Sukan, 1992; Berovič, 1992).
Antifoams should be non-toxic to humans (i.e., without handling
hazards) and the environment (i.e., able to be sewered in
reasonable quantities), and especially non-toxic to the organism
being cultured (Stanbury and Whitaker, 1984, from Hall et al.,
1973; Vardar-Sukan, 1992; Berovič, 1992). They should be non-
explosive and non-corrosive to vessels (either during sterilization
or fermentation); they should have no or low flammability
(Vardar-Sukan, 1992).
Mass Transfer. There should be no appreciable adverse effect
on oxygen transfer by antifoams (Vardar-Sukan, 1992; Stanbury
and Whitaker, 1984, from Hall et al., 1973). However, carbon
dioxide transfer as well as oxygen transfer is affected by the
presence of antifoams with changes in the carbon dioxide
removal rate causing changes in pH (Koch et al., 1995).
Downstream Isolation. There should be little or no interfer-
ence with subsequent product isolation or purification (So-
lomons, 1967; Stanbury and Whitaker, 1984, from Hall et al.,
1973). They should be non-reactive with the product and not
impart any odor or color to the isolated product (Vardar-Sukan,
1992).
Assay. Selection of a specific antifoam also can be strongly
influenced by whether a straightforward and reliable assay is
available for its detection downstream or if a suitable assay can
be developed readily. Two example methods are published: a
reverse phase LC method with evaporative light scattering
developed for simethicone (Moore et al., 2002) and a super-
critical fluid chromatography method developed for UCON-
LB625 (Dehaven et al., 1997).
Sterilization. Antifoams should be stable during sterilization
and able to be reliably steam-sterilized (Vardar-Sukan, 1992;
Stanbury and Whitaker, 1984, from Hall et al., 1973). Interest-
ingly, oil-based antifoams (e.g., polyalkyleneglycols, PAG)
require longer sterilization times than silicon-in-water emulsion
(SE) antifoams (Solomons, 1969). The early literature describes
a variety of sterilization conditions for antifoams: (1) steriliza-
tion at 160 °C for 2-3 h under dry heat conditions versus 127
°C for 90 min versus direct steam for 2-3 h at 127 °C, all
successfully applied (Bungay et al., 1960), (2) autoclaving for
90 min at 125 or 127 °C (both temperatures cited, 127 °C
corresponded to 1.5 atm pressure) sufficient for 2 L quantities
but dry sterilization at 160 °C for 3 h inadequate (Paladino et
al., 1954), (3) sterilization of larger than laboratory amounts
routinely conducted by live steam injection at 2 atm pressure
(Paladino et al., 1954). Small amounts of condensate formed
during autoclaving or provided by live steam are deemed
sufficient to provide the moist heat environment necessary for
microorganism destruction at lower steam sterilization temper-
atures (Paladino et al., 1954).
Sterilization of antifoams to be added to fermentations mid-
cycle can be difficult, especially when they are undiluted, owing
to low vapor pressures and despite sometimes lower spore
D-values in antifoam relative to water (Junker et al., 1999).
[Specifically, the D-values of Antifoam C, UCON LB625, and
P2000 relative to water are 0.8, 0.154, and 0.865, respectively
(Junker, 2001).] Interestingly, some antifoams are designed to
be added post-sterilization (i.e., they are not stable at sterilization
temperatures) which means a second antifoam can be required
during batching and sterilization. In other cases, antifoams (e.g.,
UCON HB) can be designed to become less water soluble as
temperature increases (reverse solubility), thus becoming more
effective antifoams in applications such as boiler feedwater
(Currie, 1953). However, these novel antifoams are only
transferable to fermentation applications if they are biocom-
patible.
General Trends in Application. The selection of the
appropriate antifoam agent for a specific fermentation applica-
tion must be “treated with caution and understanding” (Duitschae-
ver, 1988). No single antifoam has all the properties for all foam
776
control situations, and antifoam selection mostly is accomplished
by trial and error (Solomons, 1969). Often the only available
method of selection is experimentation (Berovič, 1992), making
empirical tests of antifoams necessary for each individual system
(Schügerl, 1985). Interestingly, antifoams capable of destroying
foam in one application could stabilize foam in another
application (Vardar-Sukan, 1991).
Only a few authors indicate trends in antifoam usage
depending on the fermentation process. Oily liquids such as
polyglycols are most effective in fungal fermentations, and
silicone-based antifoams are most effective in bacterial fermen-
tations (Solomons, 1969). Specifically, PPG is effective at 0.2%
in a fermentation of Penicillium sp., but silicone emulsion is
not; PPG is not effective in a bacterial fermentation, but silicon
emulsion is effective (Solomons, 1967). Silicone defoamers
generally are not satisfactory in mold fermentations because they
are inactivated by molds (Ghildyal et al., 1988; Solomons, 1967).
poly(dimethylsiloxane)s are most suitable for bacterial fermenta-
tions at alkaline pH and for yeast fermentations regardless of
pH (Ghildyal et al., 1988). Often only 10 ppm or less of a poly-
(dimethylsiloxane) is needed to be effective compared with 100
to 1000 ppm for lower cost organic antifoams (Ross, 1967).
Soybean and lard oil antifoams disturb sugar utilization for
sodium gluconate fermentation by A. niger, but octanol in ethyl
alcohol does not (Blom et al., 1952). Finally, inert non-
metabolizable antifoams are preferred for enzyme production,
but metabolizable antifoams are favored for secondary metabo-
lite production (Schügerl, 1985).
There are some but comparatively few reports of antifoam
use in polysaccharide fermentations or plant cell cultivations,
presumably owing to the high viscosity of these broths, which
reduces foaming. Antifoams used in plant cell culture are
sometimes used to control formation of crusts that prevent broth
circulation (Bond et al., 1987). Silicon antifoam toxicity varies
according to the type of plant culture (Bond et al., 1987).
Similarly, only relatively few authors directly compare
antifoams for their effect on production and other fermentation
parameters (Table 5). These studies are summarized with the
antifoam class listed according to Tables 2, 3, and 4 to facilitate
study comparisons. In some cases, antifoams of the same class
have substantially different effects, suggesting that the propri-
etary nature of antifoam composition can be an important
influence. In other cases, the antifoam class uniquely impacts
performance. When such studies are undertaken, some utilize
fermenters, whereas several others utilize shake flasks. Generally
small amounts of most antifoam agents increase yield over no
antifoam addition in shake flask experiments since surface
oxygen transfer is presumed higher when no foam was present
(Stefaniak et al., 1946). It can be difficult to apply shake flask
results to predict bioreactor performance, however, since no
sparging is present in shake flasks. One interesting study
attempts unsuccessfully to relate the fatty acid composition of
natural oils to its foam suppression capabililty (Table 6).
Consequently, there does not appear to be known methods to
predict an antifoam’s performance on the basis of its chemical
composition.
A tabulation of the antifoam class and specific antifoam type
is attempted for various fermentation process types to determine
if the fermentation literature suggests any further trends in
antifoam selection (tabulation details not included). The most
recently published similar tabulation of which this author is
aware is nearly 20 years old (Ghildyal et al., 1988). Several
factors make this analysis challenging. Many antifoam product
lines have changed vendors more than once, adversely impacting
Biotechnol. Prog., 2007, Vol. 23, No. 4
traceability. Often a single trade name may encompass several
types of antifoam of differing chemical composition so that the
product number becomes the critical differentiating factor. The
experimental sections of literature reports often have insufficient
information regarding the antifoam employed (e.g., concentra-
tion, vendor, trade name and product number, chemical com-
position), and in some cases, antifoam is not used at small scale
(shake flasks or stirred tank bioreactors less than 20 L), or if it
is used, its description (e.g., type, amount, and addition
conditions) is not specified. In other cases, antifoam is not
explicitly reported as a fermentation ingredient, although it
appeared likely used from the experiment’s context. Since in
various organizations (e.g., academic, industrial, government),
typically one main antifoam is used for processes of the same
nature, only one literature report from each group for the same
cultivation system was included in the analysis. Antifoams used
with carriers (Berovič, 1992) or antifoams used synergistically
for a greater combined effect (either mixed before addition or
added alternately) than when used individually (Berovič, 1992)
also are noted.
A sufficient number of published reports are examined to
determine if any trends in class of antifoam usage exist as
summarized in Table 7. Regardless of culture type, for complex
media, a majority of antifoams utilized are PAG, with silicone/
silicone emulsions forming a second significant grouping. For
semi-defined media, both PAG and silicone/silicone emulsions
are most commonly selected, although fewer studies are
conducted using this media type. For defined media, primarily
PAG antifoams are utilized with far fewer selections of silicone/
silicone emulsions. By far the most reports were collected for
lab-scale bioreactors, although more studies are reported for pilot
and production cultivations using complex media than other
media types.
In addition, a sufficient number of reports are examined where
possible to acquire enough data to determine if any trends in
the amount of initial antifoam usage exist as summarized in
Table 8 using the same set of reports used for Table 7. Initial
antifoam amounts are greater for fungal and filamentous cultures
grown on complex media compared with semi-defined and
defined media. For yeast, single cell bacterial and animal/other
cells, no such difference based on medium type is apparent.
Antifoam often is added initially to prevent foaming during
media preparation, transfer, and sterilization, as well as during
cultivation. Since foaming would be expected to decrease at
laboratory scales, initial amounts likely might be skewed by
the large fraction of laboratory reports in the set (Table 7).
Types of Antifoams. Silicone-Based. Silicones are poly-
(dialkylsiloxane)s, most commonly poly(dimethylsiloxane)s
(PDMS). Their typical viscosity is 60,000 cSt, and their typical
average molecular weight is 270,000 (Moore et al., 2002; Hall,
1971). They are virtually insoluble in water, possessing low
volatility (Hall, 1971). Their surface tension is about 21 dyn/
cm, and their interfacial tension is about 42 dyn/cm (Hall, 1971).
These properties translate into low spreading coefficients and
low dispersion in foaming systems, making them less effective
when used in their pure form (Hall, 1971).
Consequently, silicone-based antifoams contain finely divided
solids, specifically silicas, creating high surface areas and an
emulsifying agent to aid component distribution (Flannigan,
1984). Some authors feel that hydrophobic, highly dispersed,
solid silica particles are the main active ingredient and must be
present for silicone antifoams to break foam (Ghildyal et al.,
1988). Simethicones are a complex mixture of high molecular
Biotechnol. Prog., 2007, Vol. 23, No. 4 777

Table 5. Effect of Antifoam Selection and Concentration on Production and Other Fermentation Parametersa
antifoam
process class specific antifoam parameter (units) amount scale reference
bioconversion PAG P2000 0 (titer, % control) 1 g/L SF Chartrain et al., 1990
of avermectin to SE FD62 91
27-OH avermectin PAG Pluronic L 122 23
by Nocardia PAG Pluracol P2010 18
polyoxamine Tetronic 901 21
NO lard burning oil 61 5 mL/L SF
Proflo oil 21
NO FD62 330 (titer, U/L) 15 g/L 14 L BR
NO lard burning oil 410 5 mL/L
lipase production N/A control 100 (titer, % control) 0 mL/L SF Marcin et al., 1993
by Pseudomonas SE FD 62 100 5 mL/L
aeruginosa S SAG 471 127
UK SAG 4130 94
PAG P2000 102
PAG SAG 5693 100
SE Mazu 210 S 94
SE Chemax DF-10 92
SE Chemax DF-30 102
NO cod liver oil 89 1 mL/L
corn oil 22
olive oil 89
soybean oil 80
lard oil 58
citric acid N/A control 56.8 (titer, g/L) no concn 1 L BR Kamal et al., 1999
production by S silicone oil 57.5 given
Aspergillus NO groundnut oil 73.8
ALC octadecanol 55.9
sodium gluconate NO soybean oil 0.12 (% glucose utiliization/h) 0.4 mL/L 1100 L BR Blom et al., 1952
by Aspergillus NO lard oil 0.04 0.15 mL/L
ALC 1% octanol in 0.86 15 mL/L
ethyl alcohol
anti-parasitic N/A control 17 (titer, mg/L) 0 mL/L SF Burg et al., 1979
production byStreptomyces PAG P2000 53 2.5 mL/L
anti-infective N/A none 90 (peak titer, mg/L) 0 SF Stefaniak et al., 1946
production by NO lard oil 112 1 mL/L
Penicillium NO soybean oil 98
ALC 3% octadecanol 95
in soybean oil
N/A none 97 0
ALC 3% octadecanol 101 1 mL/L
in lard oil
UK Nopco (type N/A) 121
NO corn oil 142
antifungal PAG P2000 -64 (titer, ( % max) 1 mL/L SF Sigmund and Hirsch, 1998
production by SE FD-62 +<10
Micromonospora S SAG-471 -<10
PAG UCON-LB625 +40
SE MAZU-DF201 -15
SE A/Sigma -<10
SE B/Sigma +30
SE C/Sigma +13
PAG-based Antifoam 204 +<10
dispersion
PAG UCON-LB625 47.6 (titer, mg/L) 0 mL/L SF
42.7 1 mL/L
69.1 3 mL/L
93.8 4 mL/L
PAG P2000 0 (titer, ( % max) 0 mL/L SF
-49 0.5 mL/L
-87 1.5 mL/L
-95 2.0 mL/L
baker’s yeast SE not given 100 (cell mass, g/L) 0.1 g/L not given Yasukochi, 1993
(cell mass) PAG 119
PAG 121
NO 108
recombinant PAG PPG 2000 0.004 (titer, U/L) 1 g/L 2 L BR Koch et al., 1995
protein production S S184 0.0019 2 g/L
by E. coli PAG/S VP1133 0.00224 1 g/L
SE SE9 0.00048 5 g/L
biopesticide PAG PPG 10,374 (titer, IU/µL) AAN 10 L BR Vidyarthi et al., 2000
production by SE A 11,690
Bacillus NO canola oil 12,348
peanut oil 13,020
olive oil 11,946
soybean oil 11,886
778 Biotechnol. Prog., 2007, Vol. 23, No. 4

Table 5. (Continued)
antifoam
process class specific antifoam parameter (units) amount scale reference
antibody SE w/PAG C plus 0.2% w/v 37 (titer, mg/L) 0.01 g/L 2 L BR Zhang et al., 1992
production by Pluronic F-68 40 0.05 g/L
hybridoma in 45 0.1 g/L
serum- containing 48 0.2 g/L
medium
human cell N/A control 1.0 (normalized viable cell) 0 ppm SF Meis et al., 2004
line (E1A SE A 1.6 100
complementing) SE B 2.15 100
SE SE15 1.6 200
PAG P2000 1.6 1000
PAG LB-625 0.3 1000
FE O-30 0 400
plant cell culture N/A control 12 (titer, g/L) 0 SF Wongsamuth and Doran, 1994
of Atropa SE C 12 0.3 g/L
PAG PPG 1025 12
PAG Pluronic PE6100 7.5
plant cell culture N/A control 4.2 (g/L dcw) 0 ppm 8 L BR Bond et al., 1987
of Catharanthus SE 1510 8.7 0.1 g/L
N/A N/A 2.8 0 ppm
PAG P2000 10.6 (g/L dcw) 0.1 g/L
plant cell S DC-A 99.4 (relative growth) 0.1 g/L SF Wang and Staba, 1963
culture of Mentha SE DC-B 99.3 0.1 g/L
SE FD-62 99.5 0.1 g/L
SE GE-60 98.2 0.3 g/L
UK Troy-333 99.1 0.1 g/L
NO safflower 109.7 0.1 g/L
a 15,000 ppm of FD62 required upon scale up to 14 L to control foam (appearing upon bioconversion substrate addition) which lowered titer. Lard oil

(5 mL/L in medium charge and 6 mL/L added mid-cycle) used for 500 L fermentations with aeration rate of 50 Lpm (Chartrain et al., 1990). Abbreviations:
PAG, poly(alkalene glycol); S, silicone (100%); SE, silicone emulsion; NO, natural oil; ALC, alcohol; PEP, polyester polyol; FE, fatty acid or fatty acid
esters including synthetic oils; OX, oxazolidine; UK, unknown (not enough information available either from vendors or from literature); SF, shake flask;
BR, bioreactor.

Table 6. Major Fatty Acid Composition of Natural Oilsa

saturated fatty acids (%) unsaturated fatty acids (%)
natural oil palmitic stearic arachidic palmitoleic oleic linoleic linolenic
soybean 8.0 6.0 0.6 0.2 22.0 52.0 8.0
cottonseed 24.0 2.0 0.3 0.9 21.0 50.0 0.1
linseed 6.5 5.5 0 0.2 21.0 15.0 51.0
olive 9.5 2.0 0.3 1.6 81.0 7.0 1.0
sunflower 6.0 4.0 0.6 0.2 36.0 54.0 0
sesame 8.0 6.0 0.5 0.4 40.0 42.0 0.3
a Listed in decreasing order of effectiveness in suppressing foam in unsterilized medium (Vardar-Sukan, 1988).

Table 7. Summary of Antifoam Selection and Fermentation Scale for Various Culture/Media Combinations Based on Published Fermentation
Studiesa

total reports antifoam class (% reports) scale (% reports)

culture media (class/scale) S SE PAG PEP NO FE ALC OX UK flask lab pilot prod UK
fungal complex 53/59 11 9 52 6 6 4 6 6 0 7 56 29 5 3
semi-defined 7/10 0 4 4 2 0 0 0 0 0 0 70 30 0 0
defined 12/15 8 8 60 8 8 8 0 0 0 0 80 20 0 0
filamentous bacterial complex 38/42 18 13 45 5 10 0 0 0 9 7 36 48 9 0
semi-defined 5/5 20 0 40 0 40 0 0 0 0 20 40 20 20 0
defined 9/9 0 11 89 0 0 0 0 0 0 11 78 11 0 0
yeast complex 12/14 17 17 58 0 0 8 0 0 0 7 57 36 0 0
semi-defined 10/10 20 40 30 0 0 10 0 0 0 0 70 30 0 0
defined 24/28 8 8 67 0 0 13 0 0 4 0 89 11 0 0
single cell bacterial complex 40/47 13 25 55 5 0 0 0 0 2 2 79 15 2 2
semi-defined 19/19 16 21 42 5 0 0 0 0 16 0 100 0 0 0
defined 34/40 6 3 85 0 0 0 0 0 6 2 73 18 5 2
animal and other cells complex 3/3 0 67 33 0 0 0 0 0 0 0 100 0 0 0
semi-defined 1/1 0 100 0 0 0 0 0 0 0 0 0 0 0 100
defined 3/3 0 67 33 0 0 0 0 0 0 0 33 67 0 0
a Higher number of scale reports due to some reports containing studies at multiple scales. Reports that compare multiple antifoams in Table 5 are omitted

from this analysis.

weight PDMS oligomers such as dimethicon with particulate
silicon dioxide added (Moore et al., 2002).
Mixtures of mineral or vegetable oils directly with silicone
fluids and finely divided solids prove difficult to maintain
homogeneous conditions prior to use (Flannigan, 1984). Con-
sequently, finely divided silica particles are dispersed in PDMS
oil-in-water emulsions of 10-30 vol % (Solomons, 1967),
formulated to overcome problems with diluting antifoams
suitably for addition to fermentations (Keill et al., 1976). The
end result of this emulsification with surfactants and stabilizers
is to raise hydrophilicity and improve dispersibility (Hall, 1971).
Water-based silicone emulsions can be less effective in control-
Biotechnol. Prog., 2007, Vol. 23, No. 4 779

Table 8. Summary of Antifoam Amount for Various Culture/Media Combinations Based on Published Fermentation Studiesa
antifoam amount (% reports)
total e0.01% 0.01<Xe0.1% 0.1<Xe0.2% 0.2<Xe1.0% 1.0<Xe2.0% >2%
culture media reports UK AANb e0.1 mL/L 0.1<Xe1 mL/L 1<Xe2 mL/L 2<Xe10 mL/L 10<Xe20 mL/L >20 mL/L
fungal complex 54 2 13 9 31 24 29 2 0
semi-defined 9 11 11 33 33 12 0 0 0
defined 13 7 46 8 39 0 0 0 0
filamentous complex 36 0 22 14 42 8 14 0 0
bacterial
semi-defined 5 0 20 0 60 0 20 0 0
defined 9 0 22 11 45 11 11 0 0
yeast complex 11 0 27 28 45 0 0 0 0
semi-defined 10 0 20 30 50 0 0 0 0
defined 26 0 30 15 35 8 8 0 4
single cell complex 40 5 30 25 30 5 3 0 2
bacterial
semi-defined 19 10 37 10 38 0 5 0 0
defined 35 6 34 17 40 0 3 0 0
animal and complex 3 0 0 33 67 0 0 0 0
other cells
semi-defined 1 0 0 0 100 0 0 0 0
defined 2 0 0 100 0 0 0 0 0
a Reports that compare multiple antifoams in Table 5 are omitted from this analysis. b Added as needed.

ling foam than pure silicone, although the emulsions can be
readily added to fermentations (Keill et al., 1976). These
emulsions require some form of mixing to avoid settling during
prolonged stagnant periods with settling occurring faster at
higher dilutions. In addition, silicone emulsions tend to demul-
sify and break up after prolonged sterilization (Ghildyal et al.,
1988; Solomons, 1967).
PDMS-organic copolymers have been created containing
PDMS (MW g 2000) and organic PAG (polyoxypropylene of
MW g 800 and polyoxypropylene/polyoxyethylene of MW g
1500) components (Keill et al., 1976).
Polyethers. Polyethers typically are composed exclusively of
ethylene or propylene oxides or random and block copolymers
of ethylene and propylene oxides to achieve a greater range of
surface activities (Hall, 1971). Polyalkylene glycols are poly-
ethers of molecular weights typically in the range of 400-2500
(although the molecular weight of Pluronic F68 is 8400, which
may account for its more modest effectiveness as an antifoam),
with sufficient molecular weight to decrease solubility and thus
control foam effectively (Dow; Lee et al., 1993). These
antifoams have low solubility and reach gas-liquid interfaces
by diffusion (Lee et al., 1993). Specifically at 20 °C, poly-
(propylene glycol) solubility in water is 15.5 g/L and polypro-
pylene/polyethylene (copolymer) solubility in water is 3.4 g/L
(Pollard et al., 2006). Lower molecular weight polymers are
too water-soluble for foam control (Hall, 1971). Viscosities of
polyethers with molecular weights of 2000 are about 340 mPas
(Bayer AG).
One interesting report describes use of a mixture of PPGs of
various molecular weights. Specifically, PPG1025 (BDH), PPG
2025 (BDH), and FoamMaster PPG (Henkel) are used in a ratio
of 2:2:1, diluted in water about 25% v/v, and then added to a
fungal Fusarium fermentation in defined medium to final
concentrations of 0.4-0.7% v/v (Wiebe et al., 2001). Another
report describes dispersion of larger amounts of PPG using a
non-ionic surfactant, an “all-in-one” formulation, which “solu-
bilizes” the PPG (e.g., EEA-142 Henkel-Nopco) (Lee et al.,
1993).
Natural Oils. Natural oils are the earliest antifoams (Hall,
1971), serving as one type of water-insoluble organic liquid
(Vardar-Sukan, 1992). These oils generally are esters of glycerol
with long chain saturated or unsaturated monobasic acids
(triglycerides) along with some free fatty acids (Hall, 1971;
Gutcho, 1973). Typical compositions of natural oils are shown
in Table 6. Lard oil, composed mainly of glycerides of oleic
and stearic acids, is one frequently used natural oil antifoam
(Rolinson and Lumb, 1953). Recently, vegetable oils often are
selected for products with food applications since they can be
made kosher and are non-animal-sourced. Oils used as antifoams
tend to oxidize forming aldehydes, carboxylic acids, peroxides,
and other compounds with negative effects on fermentation
performance, necessitating co-addition of non-toxic antioxidants
(Rakyta et al., 1980) or short expiry dates.
Oils increase the bubble size of foams, making them less
stable (Rols and Goma, 1991). Their low hydrophilicity limits
their spreading/dispersion abilities and their high viscosity limits
their antifoaming capabilities (Hall, 1971; Vardar-Sukan, 1992).
The best oil source for antifoam applications is defined as the
source with the lowest concentration resulting in reasonably fast
foam collapse (Vardar-Sukan, 1991). However, foam suppres-
sion (preventing/inhibiting foam) as well as foam collapse
(eliminating/breaking foam) must be considered (Vardar-Sukan,
1991). All natural oils suppress foam formation (Jones and
Porter, 1998) to some degree, depending partly on the difference
between their surface tension and that of the broth (Ross, 1967).
Natural oils are energy-containing, metabolizable nutrients
(Jones and Porter, 1998). Their addition to cultivations can
increase product yields (Vardar-Sukan, 1992) or adversely affect
cultures (Ishida and Isono, 1952), particularly for genuses (such
as Mucor, Aspergillus, and Penicillium) with high lipase
activities (Ishida and Isono, 1952). For example, soybean oil is
more efficient as a foam breaker (short-lived application) than
a foam inhibitor (long-lived application) due to its consumption
by the mold under cultivation (Weng et al., 1997).
The effectiveness of natural oils in suppressing foam varies
with medium type and state (Vardar-Sukan, 1988). For 0.5-
1.0% v/v natural oil added to unsterilized soybean flour-
containing medium, foam height decreases in the following
order: linseed > cotton seed > sesame > olive > soybean >
sunflower (Vardar-Sukan, 1992). For various natural oils added
to unsterilized soybean flour-containing medium, efficiency
coefficients (based on oil cost and the amount required to reduce
foam to a minimum height) decrease in the following order:
soybean >> cotton seed >> linseed >> olive > sunflower >
sesame (Vardar-Sukan, 1992). For various natural oils added
to sterilized soybean flour-containing medium, efficiency coef-
ficients decrease in the following order: cotton seed >>> olive
>> soybean > sunflower > linseed > sesame (Vardar-Sukan,
780
1992). Finally, when 0.1 or 0.2% v/v of any one of several
natural oils (e.g., caster, corn, linseed, poppy seed, soybean)
are added to either unsterilized or sterilized complex media
(either 5% sugar beet cosette or soybean flour), the foam
collapse time decreases from minutes to seconds (Vardar-Sukan,
1991).
Depending on fermenter agitation and aeration rates, natural
oils increase, not decrease, volumetric mass transfer coefficients
up to 90% when added at 0.25% v/v to S. cereVisiae in complex
medium, recovering some of the rate reduction observed for
amounts <0.25% v/v relative to when no oil was present (Liu
et al., 1994). Below that threshold, volumetric mass transfer
coefficients are found to decrease by up to 60%, with the biggest
jump between 0 and 0.005% v/v (Liu et al., 1994). In this yeast
fermentation, 1% v/v olive oil controls foam as well as 0.1%
v/v PPG, with the olive oil yielding a similar cell density but a
much higher KLa (Liu et al., 1994). In Hansenula cultivation
using defined medium, soy (soybean) oil is less effective as an
antifoam than Desmophen 3600; the foam does not disappear,
but rather it exhibits increased bubble sizes and becomes less
stable (Voigt and Schügerl, 1981).
Polyalcohols. Aliphatic alcohols, such as n-octanol, act as
antifoams and are attractive to use in fundamental studies since
they are comprised of a single molecular species rather than a
mixture; however, they are more volatile than higher molecular
weight polymers (Lee et al. 1993). Higher molecular weight
alcohols such as octadecyl alcohol (also known as 1-octadecanol
or stearyl alcohol) also possess antifoam properties but require
a carrier since they are waxy solids. Early on, soybean oil, lard
oil, or lard oil containing 1% octadecyl alcohol were successfully
used with complex medium containing corn steep solids for
bacterial fermentations (Dworschack et al., 1954)
Specific Foaming Situations
Literature Reports. A few specific reports describing
foaming during cultivations as a function of media composition
are available. Selection of medium components and their
concentrations notably affects foaming. In a very early study
at the 30 L bioreactor scale with several fermentation systems,
4-10-fold more antifoam was required for complex than for
synthetic media, but this comparison appears clouded since
different cultures were used (i.e., no report of the same culture
being employed for both synthetic and complex media) (Rivett
et al., 1950). Foaming during cultivations of Actinomyces in
complex medium is minimized by varying media composition,
specifically using the same components at different relative
concentrations (Soifer et al., 1974). Overall, the effect of
individual medium components on foaming requires further
study (Ghildyal et al., 1988).
In a model aqueous solution, soybean meal both strongly
foams and serves as foam stabilizer depending on its concentra-
tion with its maximum ability to stabilize foam at 5 wt % and
its foaming being lower above and below 5% (Szarka and
Magyar, 1969). Glycerol also acts as a foam stabilizer (Szarka
and Magyar, 1969). Furthermore, glucose concentrations from
1% to 22% stabilize foam in 1-3% solutions of soybean meal
(Szarka and Magyar, 1969). Foaminess of cell-free medium is
believed caused mainly by the presence of corn steep liquor
(CSL) and a linear relationship between CSL concentration and
foaminess exists (Burschapers et al., 2002).
Additional reports are available describing foaming of media
during sterilization or immediately afterward. Certain media
ingredients cause foam upon sterilization, particularly complex
or natural ingredients. The elimination of foam-forming medium
Biotechnol. Prog., 2007, Vol. 23, No. 4
constituents when possible (e.g., oat flour, tomato paste, animal
extracts, meals) is the most effective way to reduce foaming
without adding antifoam.
Autoclaving, batch, or continuous sterilization potentially
causes foaming of fermentation media (Bailey and Ollis, 1977).
During sterilization, an increase in sterilization hold temperature
from 110 to 130 °C (30 min sterilization time) of molasses-
containing media causes the foaming coefficient (foam height
scaled by the ratio of foam disintegration to formation time) to
double (Viesturs, 1982). Foaming of potato protein liquor at
pH 5.2 (Emsland-starke, GmbH, FRG; protein content 30%, ash
17%, potassium content 7%, reducing sugar 8%) increases 2000-
fold during sterilization at 120 °C for 30 min (Schügerl, 1985;
Kotsaridu et al., 1983b). For a complex medium autoclaved
without sugars (20 g/L Pharmamedia, 2.5 g/L corn starch, 10
g/L ammonium sulfate, 1.3 g/L potassium monophosphate, 15
g/L salts), foaminess is low during the subsequent fermentation;
when the same medium is autoclaved with 120 g/L lactose and
2.5 g/L glucose, foaminess is considerable (Koch et al., 1995).
Foaminess during sterilization can be a function of steriliza-
tion temperature, hold time duration, pre-sterilization pH, and
antifoam concentration. Foaminess decreases from 2000 to 60
as hold temperature decreases from 120 °C to 80, and hold
duration decreases from 30 to 12.5 min (Kotsaridu et al., 1983b).
Foaminess decreases from 737 to 66 to 45 as pre-sterilization
pH decreased from 5.2 to 4.0 to 3.0, after sterilization at 120
°C for 30 min (Kotsaridu et al., 1983b). Finally, foaminess
decreases from 2000 to 67 as antifoam (Desmophen 3600)
concentration increases from 0 to 5% after sterilization at 120
°C for 30 min (Kotsaridu et al., 1983b).
Qualitative Pilot-Scale Foam Observations. Several seed
and production media containing complex and in some cases
particulate media components were examined during and after
batch sterilization in pilot plant fermenters to qualitatively assess
foaming characteristics.
Batch sterilization is conducted by heating up medium using
jacket steam until a temperature of 95 °C is reached. At 95 °C,
the jacket steam is discontinued and sparger steam is applied.
At 105 °C, steam is applied through the other internals (such
as the inoculation and medium transfer lines) until the temper-
ature is a few degrees shy of the target hold temperature,
typically 122-123 °C. During the 40 min (seed vessels) or 45
min (production vessels) sterilization hold period, the sparger
steam remains flowing, but steam through the other internals is
discontinued. Temperature is controlled through small, manual
changes in fermenter back-pressure. At the conclusion of the
sterilization, vessel back-pressure is increased to 1.5 kg/cm2 to
control foaming, sparger air is turned on at a low flowrate,
followed by shutting off the sparger steam and applying cooling
water to the vessel jacket. Once the temperature falls to below
40 °C, operating conditions are slowly adjusted to pre-
inoculation set points.
In general, at the 600 L pilot scale, complex media have a
tendency to foam during heat up, prior to reaching the desired
sterilization hold temperature. In cases where foam is somewhat
controllable [Medium 1 ) 30 g/L Stadex 60 (dextrin, A.E.
Stalely, Tate and Lyle, Decatur, IL), 20 g/L Pharmamedia
(cotton seed flour, Traders Protein, Memphis, TN), 2.5 g/L
Amberex pH (autolyzed yeast extract, Sensient Flavors, Indi-
anopolis, IN), 7.5 g/L sucrose, 5.5 g/L cerelose (glucose
monohydrate), 2.0 mL/L P2000 (Dow, Freeport, TX)], a few
inches of foam typically appears during heat up and/or when
sparger steam is applied. When internals are applied a few
minutes later, foam decreases and often dissipates only to
Biotechnol. Prog., 2007, Vol. 23, No. 4 781

Table 9. Qualitative Observations of Foaming during Various Stages of Batch Media Sterilization for Medium 3 in 600-L Pilot-Scale Vesselsa
medium heat up observations sterilization temp hold time observations cool down observations
all components when sparger steam applied, 122-124 °C foam gradually increased despite pressure increase
foam rose to top within a (more foam) to top, then subsided within to 1.5 kg/cm2 prior to
minute; foam decreased normal next 3-5 min;lowering introducing sparger air,
significantly shortly after pressure, even in small significant foaming
(<1 min) internals applied increments, results in observed
immediate foam
all components normal foam leaving 14 in.
foam to top when sparger foam to top during first
headspace
steam applied; minimal when 10 min, then subsided
all components 60 min foam to top, then subsided
applyng internals leaving 8′′ headspace
leaving 12 in. headspace
all components minimal foam observed normal minimal foam no foam
(including 2 mL/2000)
plus 20 mL/L soybean oil
a All sterilization hold times were 45 min and temperatures were 124-126 °C unless otherwise noted. Heat up and cool down procedures were normal.

reappear, sometimes rising considerably, when internals are shut Lpm. Interestingly, about 20-30 min after inoculation, the foam
off and to foam slightly during the sterilization hold phase. Foam drops dramatically (about 16 in.). As is characteristic of stable
then increases slightly when airflow is applied or in the best foams, this foam is primarily sensitive to surface-active
situations decreases or dissipates. components believed present in the inoculum.
In less optimal cases, foaming is less controllable, rising to
the vessel sightglass (top) at various points during the steriliza- Summary
tion process. For a complex medium [Medium 2 ) 20 g/L
The desired overall characteristics of antifoams are sum-
Stadex 60, 5 g/L Pharmamedia, 3 g/L NZ-Amine A (Quest
marized and presented as a compiled, comprehensive list of
International, Hoffman Estates, IL), 2 g/L yeast extract 106
properties, from which the most critical attributes must be
(autolyzed, BioSpringer USA, Minneapolis, MN), 5.5 g/L
selected for the specific process of interest. Consequently, trends
cerelose, 2 mL/L P2000], increasing the pre-sterilization P2000
in antifoam usage remain as general guidelines, based on
concentration to 6 mL/L or adding 10 mL/L soybean oil prevents
experiences of specific practitioners for their scale and cultures
foam rising to the top when internals are applied. Neither of
of interest. Despite the plethora of available antifoams, certain
these solutions is amenable for downstream processing, however.
antifoam classes more or less consistently are used for specific
In contrast, little effect is observed when 10 mL/L silicone oil
types of cultivations. Recent data from screening of alternative
(Dow-Corning 200, 10 cSt) is added. For another complex
antifoams at the shake flask and laboratory bioreactors also is
medium [Medium 3 ) 37 g/L Stadex 60, 5 g/L soy flour
presented, illustrating the process-specific nature of antifoam
(Thumb Oilseed, Ubly, MI), 7.9 g/L Hy-case (casein hydroly-
selection based on the desired process quantities for optimiza-
sate, Sheffield, Norwich, NY), 8.5 g/L yeast extract 106, 3.2
tion. By far the majority of foaming and antifoam studies are
mL/L P2000], foam is greater during cooldown for 60 min vs
conducted at the laboratory scale with significant attention
45 min sterilization hold times, consistent with more extensive
seldom devoted to finding an adequate scale down model for
Maillard reactions. In contrast to higher foam expected from
foam generation and prevention exploration. Some studies
greater Maillard degradation at higher temperatures, foam
addressing comparisons of antifoams and selection of medium
actually is lower during the sterilization hold time when the
components to minimize foam are conducted in ways that mimic
sterilization temperature is 124-126 °C versus 122-124 °C,
large-scale performance. Until further studies are available, the
presumably due to the higher vessel back-pressure of 1.3-1.35
strongest approach remains to reformulate media to minimize
kgf/cm2 required to attain the higher temperature. Table 9
or omit foaming components for processes that cannot tolerate
summarizes foaming observations during batch sterilization for
high foaming, high contamination rates, and/or high additions
Medium 3.
of antifoams.
In another complex medium at the 600 L scale [Medium 4
) 20 g/L Stadex 60, 3 g/L NZ amine A, 2 g/L yeast extract, Notation
5.5 g/L cerelose, 2 mL/L P2000] foam is watery/soapy in nature,
with about 6 in. appearing when sparger air is applied, a surface area per unit volume, 1/cm
disappearing when internals are applied, and then re-appearing DO dissolved oxygen, % sat
when internals are shut-off. A few inches remain during the KLa volumetric mass transfer coefficient, h-1
sterilization, there is no increase when sparger air is applied to OTR oxygen transfer rate, mmol/L-h
end the sterilization, and foam decreases as the batch cools, OUR oxygen uptake rate, mmol/L-h
possibly because the watery/soapy foam responds well to the Abbreviations
increased back-pressure applied during cooldown. Increasing
the PAG antifoam (2, 4, 8, and 16 mL/L) does not appreciably AAN added as needed
decrease foam; in fact foaming when air is applied to the AF antifoam
medium containing 16 mL/L is 2- to 3-fold greater. AFA antifoam agents
In another complex medium at the 15,000 L scale [Medium ALC alcohol
5 ) 6.5 g/L soy peptone SL (Marcor, Carlstadt, NJ), 6.5 g/L BR bioreactor
yeast extract 106, 5 g/L Hy-case SF, 2 mL/L P2000], foaming CSL corn steep liquor
is not problematic during sterilization. However, a stable foam DF defoamer
appears when the medium is cooled below 40 °C, which does EO ethylene oxide
not decrease with higher back-pressure or agitation rate but rises FCO foam control agent
about 6 in. when the airflow rate is raised from 1000 to 2000 FE fatty acid or fatty acid ester
782
NO natural oil
OX oxazolidine
PAG poly(alkylene glycol)
PDMS poly(dimethylsiloxane)s
PEP polyester polyol
pI protein isoelectric point
PO propylene oxide
PPG poly(propylene glycol)
S silicone (100%)
SAFD stirring as foam disruption
SE silicone emulsion
SF shake flask
UK unknown
XFO X-foam
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Received January 30, 2007. Accepted May 14, 2007.
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