Introduction
Microbial fermentation at a high cell concentration is
an effective strategy for improving the productivity of
useful solvents such as ethanol. Several methods are
usually employed to achieve high cell concentrations in
fermentation systems. These include: immobilization
techniques (Chibata et al., 1986; Amin and Doelle,
1990), repeated-batch fermentation, and cell recycle
(Scott, 1983) involving the use of separating devices
such as hollow fiber filter modules, centrifuges and floc-
culating strains of microorganisms (Ghommidh and
Bu’Lock, 1988; Hughes et al., 1990).
Hollow fiber membranes are self-supporting porous
polymer tubes which have cylindrical channels. They
offer one of the most cost effective ways of separating
cells from culture media without cell damage.
Recirculation velocity across the filter surface is of great
importance to the overall filtration rate. Maintaining a
high cross-flow rate reduces filter blockage allowing
high flux rates. Hollow fiber filter was utilized for
ethanol production by Zymomonas mobilis (Charley et al.,
1983). Earlier work utilized a microporous cassette-type
system for recycle (Beaton, 1980; Chang et al., 1981),
however difficulties were encountered in long-term
continuous cultures due to membrane and support
screen blockage.
Zymomonas mobilis (Doelle et al., 1993) is a promising
agent for large scale production of ethanol because of
its unusual physiological and biochemical properties
(Viikari, 1988) as well as its high efficiency in ethanol
production. In contrast to the exhaustive information
available on yeast fermentation, much less work has been
focused on the potential of Zymomonas mobilis for
industrial ethanol production.
Ordinary chemostat type cultures have inherent disad-
vantages which makes them unsuitable for long term
culture of anaerobic microorganisms. Unlike an aerobic
culture, economic fermentation cannot be achieved by
a reduction in the flow rate, as low sugar levels leads
to nutrient starvation and loss of viability. The chemo-
stat can therefore not be run close to the μ
max
. An
arrangement of two or more fermenters in series however
offers certain advantages over the use of a single chemo-
stat, and theoretical models for these arrangements have
been proposed (Pirt, 1975) but have not been used in
experimental setups because of constraints with equip-
ment. With biomass feedback maximum output rate of
biomass and products in a fermenter can be increased
in direct relationship to the concentration factor.
The aim of the series of experiments was to develop a
continuous culture system with high dilution coupled
with high productivity without the risk of cell washout.
Materials and methods
Microorganism and growth medium
Zymomonas mobilis NRRL-B14023 was grown on
consisting of 100 g/l glucose, 10 g/l yeast extract, 1 g/l
KH
2
PO
4
, 1 g/l (NH
4
)
2
SO
4
, and 0.5 g/l MgSO
4
.7H
2
O.
Composition of fresh medium feed was the same as the
medium employed as growth medium.
Inoculation and preculture
Z. mobilis was first propagated at 30°C for 18 h without
agitation by transferring 0.5 ml of culture from the
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Biotechnology Techniques, Vol 11, No 8, August 1997, pp. 537–541
© 1997 Chapman & Hall Biotechnology Techniques · Vol 11 · No 8 · 1997 537
Continuous culture systems involving
Zymomonas mobilis and a hollow fiber
filter module
Zakpaa Domakyaara Hilary and Ayaaki Ishizaki*
Lab. of Fermentation and Microbial Technology, Department of Food Science and Technology, Faculty of Agriculture,
Kyushu University 46-09 Hakozaki, Higashi-ku, Fukuoka-shi-812, Japan
Culture systems involving the use of one or two fermenters in series, with cell recycle employing hollow fiber filter
modules to achieve high cell densities, were developed. Dilution rates of about 0.6 h
–1
and ethanol productivity values
of about 25 g/l h
–1
, with residual glucose at 0.9 g/l, could be achieved using the system employing one fermenter.