JOURNAL OF FERMENTATION AND BIOENGINEERING

VOI. 68, NO. 2, 112-116. 1989

Comparison of Airlift and Stirred Reactors for Fermentation

with Aspergillus niger
MICHAEL TRAGER, 1 GHULAM N. Q A Z I , 2. U L F E R T O N K E N , 1. AND C H A R A N J I T L. C H O P R A 2
Universitiit Dortmund, Lehrstuhl Technische Chemie B, Postfach 500500, D-4600 Dortmund 50, FRG, 1 and
Regional Research Laboratory, Canal Road, Jammu Tawi 180001, India 2
Received 14 November 1988/Accepted 20 May 1989

The design and fabrication of a simple laboratory airlift fermentor are described. This reactor excels in
reliability and low power requirement. Its efficiency has been proved for the production of gluconic acid by
Aspergillus niger. As in the case of bubble columns, the desired morphology of A. niger, i.e pellet growth, is
obtained in the laboratory airlift fermentor as well as in a pilot scale airlift fermentor of 260 1 working volume.
Thus, pellet formation occurred under rather low stress conditions as compared to stirred tanks where high
agitation conditions are expected to be required to achieve similar results.

Gas-liquid reactors should achieve efficient mixing of
reactor contents and intensive contact o f the two phases.
Usually the gas is dispersed into the liquid phase. In the
classical type of gas-liquid reactor, the stirred tank, this is
accomplished by an agitator, but the bubble column with
its various modifications does not need a mechanical
device for dispersing the gas phase. Here contacting of gas
and liquid is done by feeding the gas stream through
distributing devices, such as perforated plates or nozzles.
In airlift reactors as a special type of bubble columns, a
circulating flow of liquid is p r o d u c e d by aerating only the
riser section separated from the rest o f the reactor
(downcomer) by a vertical wall. The simplest construction
for such an airlift reactor is the internal loop (Fig. 1). It
is a column-like vessel inside which a draft tube has been
fixed.
The aerated liquid in the riser is at least partially degass-
ed at the top o f the reactor. Due to the difference in gas
hold-up between gas-liquid dispersion in the riser and
more or less gas-free liquid in the downcomer, hydrostatic
pressure in these two regions is different which induces a
circulating flow of liquid in the reactor loop. To achieve
efficient mixing and mass and heat transfer, airlift reactors
should be tall with a large ratio of height to diameter
(about 10) (1, 2). The ratio of the diameters of riser to that
o f the outer shell is generally between 0.6 to 0.9 (3).
Due to its rather simple design, the airlift reactor has
found interest in biotechnological processes as a fermentor
(1). On an industrial scale it is used for the production o f
single cell protein and of ethanol, and in the aerobic treat-
ment o f waste water. It has also been tested on a pilot scale
for the p r o d u c t i o n of organic acids, for b i o t r a n s f o r m a -
tions, and for the cultivation o f plant and animal cells (4-
7).
Since airlift fermentors do not require a mechanical stir-
rer, the risk o f c o n t a m i n a t i o n and energy d e m a n d are con-
siderably reduced. Airlift fermentors require only about
one third o f the energy needed for stirred tank reactors.
The turbulence o f two-phase flow in airlift fermentors not
only produces favourable conditions for mass transfer, it
can also be used for suspending solid particles. Recently it
has been shown that it is possible to suspend solid particles
* Corresponding author.
112
with a density o f 2800 kg m 3 even at a solid content of
309/oo (w/w) in airlift reactors (8). This shows that airlift
fermentors are well suited for cultures containing solid
components or aggregating cells.
It was the aim o f this work to investigate whether
Aspergillus niger can be cultivated sucessfully in airlift
fermentors for the production of gluconic acid and
whether this type of bioreactor shows advantages for this
process c o m p a r e d to stirred fermentors. Since A. niger is
usually growing as mycelium, it was o f special interest to
find out whether this microorganism would show different
growth forms in the airlift fermentor c o m p a r e d to the stir-
red tank, preferably pellets instead o f mycelia.
Growth and product f o r m a t i o n in bubble and airlift col-
umn reactors have already been investigated with different
microorganisms (see Table 1). Especially in fungal fermen-
tations, where the economy of the process is dependant on
the form of growth (9, 10), i.e. mycelial or pellet growth,
these unstirred reactors have been used successfully. For
penicillin fermentations with Penicillium chrysogenum
K6nig et al. (9) as well as G b e w o n y o and W a n g (10)
reported an increase in kLa by a factor o f about 4, if the
culture grew in small pellets instead of as a pulpy
mycelium. These authors observed that pellet growth
could be achieved in bubble column fermentors by using a
preculture grown under defined conditions (9) or by im-
mobilization o f the organism (10).
MATERIALS AND METHODS
Construction o f the laboratory airlift fermentor A
rather simple design of an airlift fermentor for l a b o r a t o r y
use was given by Kiese et al. (11). This fermentor is made
from glass and has been used successfully for fermenta-
tions with various cultures (11-14, 17). Based on this
design, a stainless steel airlift fermentor was developed for
fermentations at elevated pressure which can be used for
batch and continuous cultivations (13).
The l a b o r a t o r y airlift fermentor described here has been
designed as an airlift reactor with a concentric draught
tube for a working volume of 4 . 0 / w i t h essential dimen-
sions similar to those given by Kiese et al. (11).
It consists of three parts, i.e. the conical b o t t o m holding
the gas sparger, a cylindrical part containing the draught
VoL 68, 1989 GLUCONIC ACID FERMENTATION WITH A. NIGER 113

G[1S
rh
degassing
z one

O( 0 (JO ( - ] I i

)ooo
O0
m
i

000 0
0 3 0
go
0 0 jacketed glass
0 b
0 0
cytinder
0
Oo oO
O
O O
O O O

tJ%°tJ
i

Gas
FIG. 1. Airlift fermentor with inner loop.
tube, and the top part with the degassing zone (see Fig. 2).
F o r measuring p H , temperature, and dissolved oxygen and
for removal o f exhaust gas and addition o f nutrients and
neutralization agents, several ports are provided in the
fermentor top. The b o t t o m of the fermentor holds a port
for sampling. A l t h o u g h the top and b o t t o m parts are made
o f stainless steel (AISI 304), the cylindrical part o f the
fermentor is a glass tube with a glass jacket. By circulating
water through this jacket the temperature o f the fermen-
tors contents can be controlled. The fermentor can be
steam-sterilized in an autoclave.
Dimensions of the fermentor are given in Table 2. The
conical b o t t o m o f the fermentor with the sampling port
welded to it consists o f two parts between which the gas
sparger plate is tightened by an O-ring and two teflon ring-
gaskets. The diameter and configuration o f the perfora-
tions on the sparger plate (AISI 304) have been calculated
according to Ruff et al. (15) to achieve homogeneous
dispersion o f the gas phase. The design o f sparger b o t t o m
(60 ° cone) has also proved suitable for suspending solid
particles (8, 12, 21). The distance between the draught tube
and reactor wall is s,Jfficient for installing p H - and oxygen-
probes via the ports in the reactor top. The diameter of the
top part is enlarged to reduce the liquid velocity, thus pro-
moting degassing of the liquid before it enters the
downcomer. The cylindrical part is tightened by two
silicon ring-gaskets and pressed by bolts and nuts in be-
7
3~ "~. spGrger bottom
with perforated plate
\1/
FIG. 2. Construction of laboratory airlift fermentor (dimensions
see Table 2).
tween the steel parts. In the design of Kiese et al. (11) this
enlargement of diameter in the top part has been ac-
complished by way o f a conical dilation. In the airlift
fermentor used in our investigation the degassing zone was
welded from two tubes of different diameter (250 and
100mm) for the sake of simplicity of fabrication. This
may lead to stagnant regions in this part of the fermentor.
Visual observation of mixing in the airlift fermentor showed
that possible effects o f stagnant regions on mixing in the
airlift fermentor can be neglected. Of course, when
suitable machinery and materials are available, conical dila-
tion o f the degassing zone, as depicted in Fig. 2, is to be
preferred.
Microorganism A s p e r g i l l u s niger RRL 12-16/1-2
strain selected at the Regional Research L a b o r a t o r y , Jam-
mu, India.
Inoculum P r o d u c t i o n medium was inoculated with
2%0 (v/v) of preculture grown for 24 h in shake flasks for
fermentation in 4-l airlift and 1 0 - / s t i r r e d tank reactors.
F o r the 2604 fermentor the preculture was grown in two 5-l
airlift glass fermentors.

TABLE 1. Fermentation systems tested in airlift (AL) or bubble column (BC) fermentor
Microorganism Process Reference Fermentor
C. utilis continuous (11) AL
P. fluorescens continuous; increased pressure (13) AL
T. ferrooxidans batch; leaching of sulfides from flotation residue (14) AL
T. ferrooxidans batch; removal of H2S from exhaust gases AL
Mixed culture of Thiobacilli and Leptospirillurn-like bacteria continuous; leaching of sulfides from flotation residue (12) AL
S. fragilis. C. blankii continuous fermentation of whey (17) AL
A. niger batch [ 18) BC
P. chrysogenum batch (9) BC
P. chrysogenum batch; immobilized on Celite beads (10) BC
114 TR)kGERET AL. J. FERMt:Nr. BIOEN(;.,

TABLE 2. Dimensions of the airlift fermentors coefficient kLa and circulation time tc were calculated for
Laboratory scale the air/water system at an air flow rate of V~=3 l/min. For
the kta the dynamic method was used, taking into account
Reactor: volume 4l the delay of the oxygen probe. The circulation time was
total length ca. 500 mm estimated with the aid of suspended polyethylene particles.
Cylindrical part:
length 250 mm For kca a value of 0.6 min 1 was obtained and for the cir-
inner diameter 94 mm culation time about 4 s. This kca-value agreed well with the
diameter of draught tube 52 mm value measured by Takafiashi and Yoshida (18). For com-
length of draught tube 350 mm parison of their resulting kLa with the kLa-value obtained
Bottom part with sparger: in our study, one must relate the gas flow rate of 3 I/min to
60° conical bottom
diameter of sparger plate 40 mm the total cross-sectional area of the airlift fermentor. With
19 circular holes (diameter 0.8 mm) our superficial gas velocity of 0.7 cm/s a kta-value of
in 8 mm triangular pitch about 0.9 rain ~is calculated for the system air/water with
Top part: 250 mm Fig. 3 from (18). Taking into consideration that the higher
diameter of degassing zone 80 mm liquid velocity in airlift fermentors compared to bubble col-
height of degassing zone umns results in a reduced gas hold-up (1, 19) and thus in a
Pilot scale (23) reduced interfacial area, our value for kLa is indeed in the
Reactor: volume 260 l expected range. In fermentation media, kLa will be further
liquid height 9.25 m reduced due to medium components such as proteins or an-
diameter of riser 150 mm tifoam agents. For our fermentations with A. niger a
diameter of downcomer (external loop) 50 mm decrease in volumetric mass transfer coefficient by about
Sparger plate: 75% can be expected (18). For the laboratory airlift
sintered metal with 5 pm mean pore size
Top part : fermentor of Kiese el al. (11) the same circulation time was
volume of degassing zone 80 / observed at the gas flow rate used in our study. These
authors (11) showed that even at rather low gas flo~ rates
of ~ = 0 . 5 l / m i n an airlift fermentor of this design and
Medium Fermentations in 4-I airlift and 10-1 stirred size can be considered as ideally mixed. Consequently, any
fermentor: 40 g/l of glucose. H:O, I g/l of NHaNO~, 1 g/l influence of distributed parameters can be neglected in the
of KH2PO~, 0.25 g/1 of MgSO4.7H:O, and 13 g/l of CaCO3 laboratory airlift fermentor used in this study.
in deionized water. The pH was adjusted to 6.5 with Gluconie acid production with A. niger The
NaOH. F e r m e n t a t i o n in 260-• airlift fermentor: 100 g/I of gluconic acid fermentations were done simultaneously
glucose. H20, 1 g/l of KH2PO4, 20 g/l of corn steep liquor, in the stirred tank and in the laboratory airlift fermentor.
and 0.25 g/l of MgSO4.7H20 in deionized water. The pH The preculture for both reactors consisted only of loose
was adjusted to 6.5 and controlled during fermentation mycelia. After about 6 h the microorganisms in the airlift
with 8 N NaRCO3 solution. fermentor turned into fine pellets but in the stirred fermen-
Fermentors Fermentations were done at 30°C both tor they continued to grow as loose mycelia. This con-
in a stirred tank fermentor ( 1 0 / o p e r a t i n g volume; New tradicts the study of Gomez el al. (20) who obtained good
Brunswick, USA) and in the laboratory airlift fermentor pellet formation of A. niger rather at high turbulence ex-
(4 l operating volume). The suitability of the airlift fermen- posing the culture in a stirred tank fermentor to a higher
tor design for fungal fermentations was also tested in an ex- stirrer speed. To confirm pellet formation of A. niger on
ternal loop airlift fermentor of 260 / working volume (see pilot scale, a fermentation was done in the 260-/external
Table 2). The air flow rate and stirrer speed in the stirred loop airlift fermentor. The production medium was changed
fermentor were 5 l / m i n and 6 0 0 r p m , respectively. Air from defined to complex medium because pellet formation
throughput in the 4-I and 260-I airlift fermentor were was expected to be influenced negatively by higher
4 l/rain and 35 l/rain, respectively. Superficial liquid velocity growth yields in the complex medium. In Table 3 data of a
was adjusted to WsL-8.4 cm/s by a ball valve situated in typical fermentation run in all fermentors are given. From
the downcomer of the fermentor. The fermentor was these data it is evident that for the fermentations in both
equipped with 9 oxygen sensors for the measurement of laboratory scale fermentors no significant difference occur-
the dissolved oxygen profile of the loop. A n inductive flow red in production of gluconic acid (ca) and consumption
meter for the measurement of liquid velocity was used in of glucose (cs). Dry cell weight (X) was also comparable
the downcomer. but slightly increased in the 260-• airlift fermentor. 111 the
Analysis Glucose was analyzed as reducing sugar by two laboratory scale fermentors a conversion of glucose to
the ferricyanide method (16). Gluconic acid in the gluconic acid of 85-900/oo (w/w) was achieved, with 10~( of
laboratory scale fermentors was measured by both the substrate remaining unused. In 260-1 airlift fermentor,
titrimetry and paper chromatography (24) after filtering the product yield was increased to about 98°~) (w/~x). In-
and decationization of the culture broth. For the pilot terestingly, the dissolved oxygen concentration was
scale airlift fermentor, acid was calculated from the distinctly higher in the airlift fermentor than in the stirred
a m o u n t of added Na2CO3 solution and subsequently con- fermentor at the end of the fermentation. This may be ex-
firmed by H P L C (Biorad column H P X 87H ~ at 30°C us- plained by the fact that, due to the small pellets in the
ing 0.4 mM H 2 S O 4 as the eluent). Dry cells were weighed airlift fermentor, mass transfer was better than in the stir-
after dissolving CaCO 3 (where present) with hydrochloric red fermentor, in which broth viscosity was rather high. In
acid, filtering, and drying. the airlift fermentor a water-like viscosity of the broth
prevailed but in the stirred reactor the mycelium grew as a
pulpy mass which is known to behave as a non-Newtonian
EXPERIMENTAL RESULTS
liquid. Viscosities were not measured.
Testing of fermentor operation The mass transfer As far as cultivation of A. niger in the 260-/ airlift
VOL. 68, 1989 GLUCONIC ACID FERMENTATION WITH A. NIGER 115

TABLE 3. Typical results of batch fermentations

a) Airlift fermentor
4-1 volume
t cs c.,, X DO
Morphology of microorganisms
(h) (g//) (g/l) (g/l) ( 9~)
0 39.0 -- -- 100
8 28.0 9.5 0.7 80
14 I 1.0 27.0 1.4 26 small pellets (1-2 ram)
20 4.0 34.5 2.3 30
21.5 4.0 34.8 2.3 17
260-/volume
t cs CA X DO ~'
Morphology of microorganisms
(h) (g/t) (g/O (g//) ( T0)
0 82.6 0 1.8 110
11 50.4 23.9 l. 1 27
13.5 28.3 42.7 2.8 13
16 23.4 52.9 2.8 13 small pellets (1-2 mm)
17 19.5 65.1 3.2 13
18 9.3 73.3 3.7 7
19.5 3.8 81.1 3.5 7

b) Stirred tank fermentor

t cs CA X DO
Morphology of microorganisms
(h) (g/t) (g/t) (g//) (%)
0 39.0 -- -- n.d.
8 27.5 10.0 1.1 n.d.
14 10.0 24.0 1.5 n.d. loose mycelium
20 5.0 32.8 2.1 5
21.5 3.0 33.0 2.1 5
a In downcomer.
n.d. not determined.

f e r m e n t o r is c o n c e r n e d , a u n i f o r m pellet g r o w t h similar to
t h a t o b t a i n e d in t h e l a b o r a t o r y airlift r e a c t o r was a c h i e v e d .
C o n s i d e r i n g the c o m p l e x n a t u r e o f the m e d i u m u s e d ,
cell g r o w t h was r a t h e r low (cf. T a b l e 3). This is q u i t e
d i s t i n c t f r o m t h e g r o w t h ( a b o u t 15 g / / ) r e p o r t e d in similar
m e d i u m in a s t i r r e d r e a c t o r o f 400 1 v o l u m e by Z e t e l a k i et
al. (22). T h u s w i t h a yield o f 98%o ( w / w ) g l u c o n i c acid
f r o m g l u c o s e , the specific activity o f A . niger in the airlift
f e r m e n t o r was q u i t e h i g h .
NOMENCLATURE
cA : c o n c e n t r a t i o n o f g l u c o n i c acid, g l t
Cs : c o n c e n t r a t i o n o f glucose,
DO : dissolved oxygen c o n c e n t r a t i o n related to satura-
t i o n with air at 1 b a r , %
k~a : v o l u m e t r i c m a s s t r a n s f e r coefficient, m i n
t : f e r m e n t a t i o n time, h
t~ : liquid c i r c u l a t i o n t i m e (time r e q u i r e d f o r o n e cir-
c u l a t i o n o f liquid p h a s e in airlift f e r m e n t o r ) , s
Vg : air flow r a t e , 1 m i n
X : dry cell c o n c e n t r a t i o n , g l i
ACKNOWLEDGMENTS
We thank Prof. F. Yoshida for discussions. Support of the col-
laboration by the Council of Scientific and Industrial Research (New
Delhi, India), International Bureau of Kernforschungsanlage (Jtilich,
F.R.G), and Fonds der Chemischen lndustrie (F.R.G.) is gratefully
acknowledged.
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