Chemical Engineering Communications

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CHARACTERISATION OF CAKE COMPRESSIBILITY

IN DEAD-END MICROFILTRATION OF MICROBIAL
SUSPENSIONS

A. A. MCCARTHY , P. GILBOY , P. K. WALSH & G. FOLEY

To cite this article: A. A. MCCARTHY , P. GILBOY , P. K. WALSH & G. FOLEY (1999)

CHARACTERISATION OF CAKE COMPRESSIBILITY IN DEAD-END MICROFILTRATION
OF MICROBIAL SUSPENSIONS, Chemical Engineering Communications, 173:1, 79-90, DOI:
10.1080/00986449908912777

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CHARACTERISATION OF CAKE
COMPRESSIBILITY IN DEAD-END
MICROFILTRATION OF MICROBIAL
SUSPENSIONS
A. A. McCARTHY, P. GILBOY, P. K. WALSH
and G . FOLEY*
School of Biotechnology, Dublin City University, Dublin 9 , Ireland

(Received I June 1998; Infinal form 27 October 1998)

Culture broths of Saccharomyces cerevisiae and KIuyveromyces marxianus var marxianus

NRRLy2415, suspensions of rehydrated active dry bakers yeast and suspensions of calcium
carbonate were filtered in dead-end mode at pressures below 200kPa. In the case of all the
microbial suspensions, the specific cake resistance was found to be a linear function of pressure.
Cake compression was found to be reversible or weakly irreversible with respect to changes in
pressure, i.e.. the specific resistance measured at a given pressure was only weakly dependent
on whether the filter cake had previously been exposed to higher pressures. The greatest irre-
versibility effects were obtained with unwashed active dry yeast suspensions, consistent with the
breakdown of cell aggregates in these suspensions. The specific cake resistance of calcium car-
bonate suspensions was found to be a non-linear function of pressure. The compression of cal-
cium carbonate cakes was irreversible, consistent with the breakdown of the large number of
particle aggregates in these suspensions.

Keywords: Microfiltration; yeast; compressibility; reversibility; aggregate

INTRODUCTION

Filtration of microorganisms is a n important step in the recovery of many

bioproducts. In recent years, filtration has been the subject of considerable
research. However most of the reported studies have concentrated on the
relatively new technique of crossflow filtration [I]. Investigations into the
older and more industrially important technique of dead-end filtration have

'Corresponding author. Tel.: +353-1-7045395, Fax: f353-1-7045412, e-mail: greg.foley@

dcu.ie
80 A. A. McCARTHY er a / .

been comparatively rare and many fundamental questions regarding the pro-
perties of microbial filter cakes remain unanswered. In particular, the mecha-
nism of cake compressibility in microbial filtration is not well understood.
Filter cake compressibility is the term used to denote the tendency for the
specific cake resistance to increase with pressure. The specific cake resist-
ance, a, can be related to the filtrate flux, J, through the equation

where AP is the applied pressure, M is the mass of solids deposited per

unit membrane area, RM is the membrane resistance and p is the filtrate
viscosity. Traditionally, the specific resistance has been related to pressure
using a n expression of the form
cr = a A P n (2)
where a and n are empirical constants. In the filtration of non-microbial
suspensions, Eq. (2) has generally been found to represent the pressure de-
pendence of the specific cake resistance a t high pressures [2]. Equation (2)
has been found to be less accurate for microbial suspensions which are
generally filtered a t low to moderate pressures ( < 200 kPa). For example,
Nakanishi e f al. [3] observed non-linearity in log-log plots of cr versus A P
and estimated the compressibility index, n, by fitting Eq. (2) to a linear
region of the plot a t the high end of the pressure range. Other researchers
have employed a similar procedure [4].
In our previous work, we demonstrated that the specific resistance of
Kluyveromyces marxianus filter cakes was a linear function of pressure [5].
Given this finding and since Eq. (2) appears to give an inadequate des-
cription of cake compressibility in other microbial filtration studies [3, 41,
the aim of the work presented in this paper was to re-examine the relation
between specific cake resistance and pressure for a range of microbial
suspensions. In addition, the mechanism of cake compressibility was investi-
gated by studying the reversibility of cake compression. The microbial
filtration data were compared with data for a model non-microbial
suspension, calcium carbonate.

MATERIALS AND METHODS

Microorganisms
The microbial suspensions used in this study were (i) resuspended active dry
yeast (Distillers Company Limited, Scotland), (ii) washed suspensions of
 FILTRATION OF MICROBIAL SUSPENSIONS 81

the same active dry yeast, (iii) the same yeast reactivated in culture medium
as described previously [6], (iv) the fungus Kluyveromyces marxianus var
marxianus NRRLy2415 (Northern Regional Research Laboratories, Peoria,
IL) which displays a variety of morphological forms, from simple ovoid
yeast to branched mycelia as described previously [7].

Fermentation
A YEPD medium consisting of 10g/L yeast extract, 20g/L bacteriological
peptone and 20g/L D-glucose was prepared for cultivation of the bakers
yeast (BY). Cultivation was performed on a shaker table (133 rpm) at 30C
for 24hours. Both yeast-like (KMI) and mycelial (KM2) K. marxianus
suspensions were grown on whey based medium, in batch and continuous
culture, as described previously [5, 71.

Resuspension of Active Dry Yeast

7.5 g/L of dried yeast was resuspended in 250mL of deionised water at pH 5.
The mixture was agitated magnetically in a IOOOmL Erlenmeyer flask at
30C for 30minutes. Suspensions prepared in this way are referred to
throughout this paper as "unwashed" suspensions (ADYI). "Washed" sus-
pensions (ADY2) were prepared by centrifuging an unwashed suspension
for 20minutes at 10,000rpm in a Sorval centrifuge, decanting the super-
natant and resuspending the pellet in a fresh volume of deionised water.

Determination of Cell Concentration

Wet cell concentrations were determined using a modified form of the meth-
od of Ju and Ho [8], as described in detail elsewhere [5].

Image Analysis
The mean specific cell surface area, S, (defined as the mean surface area
per unit volume) and mean cell aspect ratio, Lh, (defined as the mean ratio
of length to equivalent cylindrical diameter) were determined for each
microbial suspension using a Leica QSOOMC image processing and analysis
system as described previously [S]. Aggregated cells were excluded from
all measurements and no attempt was made to quantify the degree of
82 A. A. McCARTHY er a/.

aggregation. The mean morphology of cells in each suspension as

determined by image analysis is summarised in Table I.

Calcium Carbonate Suspensions

For comparison with the microbial filtration experiments, a series of filt-
rations were performed with 40g/L suspensions of CaCO,, suspended in
water in exactly the same manner as the unwashed yeast suspensions.

Filtration
Filtration was performed using a stainless steel filtration cell provided by
Gelman Sciences (Dublin, Ireland). Supor (polysulphone) membranes of
nominal pore size 0.45 pm and diameter 47 mm were used in all filtrations.
A new filter was used for each run. Pressure was maintained at a constant
value by attaching the cell to a Nitrogen cylinder. The pH of all suspensions
was adjusted to 5 by the addition of 10% NaOH or 10% HCI prior to
filtration. All filtrations were performed at 20C. Filtrate volumes were
determined at various times using a Mettler electronic balance (Mettler,
Germany) and the flux, J, was determined as A V/(AAt) where A is the mem-
brane area for filtration and AV is the change in filtrate volume during a time
period At.
After filtration of suspensions was completed, the cake was removed
and the pure-water flux was compared with the clean-membrane value. No
significant differences were observed, indicating that membrane fouling was
negligible.
The specific resistance of a filter cake, a,was determined using the method
of Nakanishi et al. [3]. A 150mL volume of suspension was filtered at a
pressure of 30 kPa. The pressure on the cake was released, the filtrate

TABLE I Cell morphology

Suspension SV Lim
(urn-') (-)

KM I (yeast-like)
KM2 (mycelial)
ADY l (unwashed)
ADY2 (washed)
BY
 FILTRATION OF MICROBIAL SUSPENSlONS 83

returned to the filter module, the pressure set at 30 kPa and filtration was
allowed to proceed. The specific resistance, a, could then be calculated from
the following expression [5]

where c is the wet cell concentration and V, is the volume of filtered sus-
pension (150mL). Accuracy in the estimation of a was ensured by allowing
sufficient filtrate to pass through the membrane until a steady-state filtrate
flux was attained. The specific resistance was measured at four further
pressures (60,100,140 and I80 kPa) by incrementing the pressure and
measuring a steady flux as before. In this manner, specific resistance
measurements over the desired range of pressures could be achieved in a
single pass through the module. It should be pointed out that after each
change of pressure a steady state flux was reached almost instantaneously
and hence no information could be obtained regarding the dynamics of the
change in specific resistance. After the specific resistance had been measured
at the highest pressure, the pressure was released, the entire filtrate was
returned to the cell and the specific resistance measured at all pressures for
a second time. The procedure followed was exactly as for the first pass.
The procedure was modified for calcium carbonate to prevent the cake
running dry and compacting [9].The filter cell was charged with 200mL of
suspension and filtration carried out at 30 kPa until such time as the flux
reached a constant value due to settling of particles. This typically occurred
when 150mL of filtrate had been produced, at which point the pressure was
released and the 150 mL returned to the filter cell. The subsequent procedure
was the same as for the microbial suspensions.

Filtrate Viscosity
Filtrate viscosities were determined at 20C using a Brookfield V1 (Harlow,
England) cone and plate viscometer.

RESULTS AND DISCUSSION

Microbial Suspensions
Figures 1 to 3 indicate the relation between specific cake resistance and
pressure (first and second passes) for the microbial suspensions.
 A. A. McCARTHY er a / .

Pressure (kPa)
FIGURE 1 Specific cake resistance versus pressure for yeast-like (KMI) and mycelial (KM2)
broths of K. marxianus. Solid and open symbols represent data for the first pass and second pass
respectively. The lines represent fits of Eq. (4) to the experimental data. In the case of KM2,
differences between the data for each pass were not great enough to be visible on the graph.

The linearity of the microbial suspension data is consistent with the

upward curvature seen in log-log plots of a versus A P in Nakanishi et 01,'s
work [3]. It is clearly not appropriate to correlate our specific resistance data
using Eq. (2). The use of this equation requires a subjective selection of
a linear region on the log-log plot and will also lead' to considerable
inaccuracies in the prediction of the specific cake resistance at low pressures.
A more appropriate equation for correlating microbial specific cake
resistance data is [S]

where a. is the specific resistance of the unstressed cake and kc is a constant

which is a measure of cake compressibility. In all cases, regression coefi-
cients for fits of Eq. (4) to our data exceeded 0.997.
 FILTRATION OF MICROBIAL SUSPENSIONS

24 s

-i

0 40 80 120 160 200

Pressure (kPa)
FIGURE 2 Specific cake resistance versus pressure for unwashed active dry yeast (ADYI) and
washed active dry yeast (ADYZ). Solid and open symbols represent data for the first and second
pass respectively. The lines represent fits of Eq. (4) to the experimental data.

The Carman-Kozeny equation predicts that the specific cake resistance is

proportional to S: [lo]. However, it does not make any prediction regarding
the effect of particle shape on specific cake resistance. In Figure 4, a plot'is
given of cro/S: versus L* for all microbial suspensions studied, where cro
has been determined from the intercept of the data fits in Figures 1-3. The
dominant trend in Figure 4 is that cro/S: decreases with increasing L,,,,,.This
is consistent with our earlier work on the filtration of K. marxianus [5]. The
reduction in the ratio ao/S; as Ldm increases is due to the increased voidage
in cakes composed of more elongated cells [ 5 ] .
Figure 4 shows that there was a difference between the first and second
pass data for all suspensions. However, the differences are very small in the
case of the culture broths, suggesting a predominantly reversible mecha-
nism of cake compression. For the ADY suspensions, however, substantial
86 A. A. McCARTHY ef a1

0 40 80 120 160 200

Pressure (kPa)
FIGURE 3 Specific cake resistance versus pressure for cultured Bakers yeast (BY). Solid and
open symbols represent data for the first and second pass respectively. The lines represent fits of
Eq. (4) to the experimental data.

differences are observed, indicating a significant degree of irreversibility in

the cake compression process. This irreversibility is particularly pronounced
for ADYI, possibly reflecting the fact that this suspension contained
substantial numbers of cell aggregates. Aggregated particles have been
shown to form cakes which have increased porosity [2], thus explaining the
particularly low value of cxO/S:for ADY 1 in the first pass. Aggregates are
also likely to breakdown irreversibly under the action of pressure, a fact
which explains the substantial difference between the first and second pass
data for ADY1. It is not possible to confirm that aggregate breakdown
occurs because the very process of recovering the filter cake and resus-
pending the cells for microscopic analysis affects the degree of aggregation.
 FILTRATION OF MICROBIAL SUSPENSIONS

ADYl
A ADY2
BY

FIGURE 4 ao/.Y; versus Lh for microbial suspensions. Solid and open symbols represent
data for the first and second pass respectively. First and second pass data for KM2 are identical.

Figure 5 is a plot of kc versus Ld, for all the microbial suspensions. As

found in our previous work [5], cake compressibility becomes more
pronounced as the aspect ratio of the cells in the culture broth increases.
The ADY suspensions, however, exhibit enhanced cake compressibility.
Given that the cell walls of rehydrated yeast are usually damaged due to
the drying process [Ill, this may lead to increased cake compressibility,
possibly because of greater cell deformability [12]. The enhanced compres-
sibility of the ADYl suspension may once again reflect the breakdown of
aggregates. It is likely that compression of ADYl cakes proceeds by two
mechanisms - a reversible mechanism similar to that undergone by the
culture broths and an-irreversible mechanism related to aggregate break-
down. This is consistent with the fact that the ADYl cakes show the greatest
reduction in compressibility from the first to the second pass.
 A. A. McCARTHY el al.

FIGURE 5 kc versus Ld, for microbial suspensions. Solid and open symbols represent data
for the first and second pass respectively. First and second pass data for KM2 are identical.

Calcium Carbonate
Figure 6 is a plot of specific cake resistance versus pressure for the calcium
carbonate suspension. The first pass data are clearly non-linear over the
range of pressures examined. The non-linearity of the calcium carbonate
data is possibly an indicator of fundamental differences between the cake
compression mechanisms of microbial and powder suspensions. This is
supported by the highly irreversible nature of the cake compression process
when calcium carbonate is employed. It is well established in the literature
that the compression of powder cakes is mainly associated with irreversible
breakdown of particle aggregates [2]. Microscopic examination revealed
large numbers of aggregates in the calcium carbonate suspensions used in
this study.
 FILTRATION OF MICROBIAL SUSPENSIONS

0 40 80 120 160 200

Pressure (kPa)
FIGURE 6 Specific cake resistance versus pressure for calcium carbonate. Solid and open
symbols represent data for the first and second pass respectively.

CONCLUSIONS

At pressures below 200 kPa, the pressure dependence of the specific cake
resistance of microbial filter cakes would appear to be best described by a
linear equation such as Eq. (4). Fitting of Eq. (2), the standard power-law
expression, is subjective and inaccurate at low pressures. At present, there
is no explanation for the essentially reversible nature of microbial cake
compression. Direct observations of changes in cell morphology under
pressurised conditions are required before any real insight into the
compression mechanism can be obtained. Irreversibility effects arising in
the compression of unwashed bakers yeast and CaC03 cakes appear to be
due to the breakdown of aggregates. Current work in our laboratory is
90 A. A. McCARTHY ef a/.

focusing on using image a n a l y s i s t o q u a n t i f y the degree of cell a g g r e g a t i o n

in suspensions of a c t i v e dry yeast. By comparing t h e degree of aggregation
with the irreversibility of cake c o m p r e s s i o n , we hope to confirm the role of
aggregates in t h e cake c o m p r e s s i o n process.

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