Pergamon

Chemical Engineering Science, Vol. 51, No. 9, pp. 1487 1494, 1996
Copyright © 1996 Elsevier Science Ltd
Printed in Great Britain. All rights reserved
0009 2509/96 $15.00 + 0.00

0009-2509(95)00305-3

D I L U T E S O L U T I O N VISCOSITY OF RED M I C R O A L G A
EXOPOLYSACCHARIDE
EDWARD ETESHOLA, MOSHE GOTTLIEB *t and SHOSHANA (MALIS) ARAD *
tDepartment of Chemical Engineering and *Department of Life Sciencesand The Institutes for Applied
Research, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel
(First received 4 November 1994; revised manuscript received and accepted 24 April 1995)
Abstract--The red microalga Porphyridium sp (P. sp) is encapsulated in a sulphated polysaccharide. The
external part of this capsule dissolvesin the growth medium. This extracellular polysaccharide is a heteropolyelectrolyte with molecular weight of ~ 6 x 106 Da. The effect of solvent, counterion and pH on chain
flexibilityand structural features in dilute solution of the exopolysaccharide was investigated by intrinsic
viscometry. From the dependence of the intrinsic viscosity [q] on ionic strength, it was estimated that the
stiffness of P. sp polysaccharide chains is in the same range as that of xanthan and DNA. The effect of the
counterion on [q] is found to be specific and dependent on the type and valence of the counterion. The
polyelectrolyte behaviour of the polymer is confirmed by the decrease of [q] with the addition of salt
without any observable order-disorder conformational transition in aqueous salt solutions in the commonly used range of ionic strength (0.01 1.0). At considerably lower ionic strength (< 0.01) there is an
indication of a transition in the P. sp polyion conformation, most likely reflecting a contraction of the
polymer chain from a highly stretched to a stiff, wormlike chain. It is hypothesized from the overall dilute
solution features that the P. sp biopolymer chain molecules adopt stiff ordered conformation in solution.

INTRODUCTION

Porphyridium sp. is a red microalga which is encapsulated in a sulfated polysaccharide. The external part
of this capsule dissolves in the growth medium. This
extracellular mucilaginous material (hereafter referred
to as PspP, the second P standing for polysaccharide/polymer) carries carboxyl and half ester sulfate
groups on its glycosidic backbone which confers on it
the properties of a biopolyelectrolyte. The exocellular
polymer of P. sp contains different sugars, including
xylose, galactose, glucose, mannose, arabinose,
methyl hexoses and methyl pentoses in various
amounts and ratios (Jones, 1962; Ramus, 1973; Medcalf et al., 1975; Heaney-Kieras and Chapman, 1976;
Heaney-Kieras et al., 1976; Percival and Foyle, 1979;
Geresh et al., 1992). In addition, the polysaccharide
contains ca 9% glucuronic acid, ca 10% half ester
sulfate, and a protein moiety (Heaney-Kieras et al.,
1976). The molecular weight of the P. sp polysaccharide was estimated as 5-7 x 106 Da (Simon et al., 1992).
The polysaccharide produced by the red microalga
P. sp is of great interest because at low polymer
concentrations it yields highly viscous aqueous solutions with unique theological properties comparable
with those of xanthan and carrageenan, and thus
suitable for technological applications (Savin, 1978;
Ramus, 1986; Ramus et al., 1989; Geresh and Arad,
1991). For example, dilute aqueous solutions of the
polymer have been shown to be effective in drag

*Corresponding author.

reduction in capillary pipe flow (Ramus, 1986; Ramus
et al., 1989). Due to its special properties, PspP aqueous solutions have been reported to be compatible
with various salts and stable under varying pH, high
temperature, and accelerated flow rates (Savin, 1978;
Ramus, 1986; Ramus et al., 1989; Geresh and Arad,
1991). Preliminary studies in our laboratory indicate
that P. sp polysaccharide has also interesting surface
activity properties and has potential use as a pharmaceutical material.
Most of the work published in the literature on this
polysaccharide is concerned with glycosyl content
and chemistry and hardly touches upon the relationships between conformational structure and rheological properties (Geresh and Arad, 1991). In order to
develop the extracellular polysaccharide of P. sp, we
need to understand their structure-function relationships, which presupposes a detailed knowledge of
their molecular behaviour and physico-chemical
properties (Lee and Chandrasekaran, 1992). Accordingly, in the work described in this paper we examined
the dilute solution properties of PspP as a function of
the nature of the solvent (water, aqueous salt solutions, pH), and the type of counterion. Our purpose
was to determine the influence of these experimental
variables on the chain flexibility and on the conformational features of the polymer in dilute solutions as
manifested by intrinsic viscosity values.
EXPERIMENTAL
Alga and growth conditions
Porphyridium sp. (UTEX 637) was obtained from
the culture collection of algae at the University of

1487
CE5 51:9-,,1

1488

E. ETESHOLA et al.

Texas, Austin. The cells were grown in batch culture
in 1-1 columns 6 cm in diameter at 24 + I°C in artificial seawater according to Jones et al. (1963). The
cultures were illuminated continuously with fluorescent cool-white lamps at an irradiance of 150 microeinsteins m - 2 s- ~. The medium was aerated with sterile air containing 3% COg (Arad et al., 1988). At the
stationary phase of growth (after 2 weeks), the cells
were separated from the growth medium by centrifugation at 10,000 rpm for 20 min using a Sorvall centrifuge model RC2-B (Sorvall Instruments Du Pont).
The supernatant medium containing the polymer was
collected and dialyzed in Visking size 98 32/32" dialysis tubing (Medicell International Ltd., London)
against distilled water and then exhaustively against
bidistilled water at 4°C. The resultant solution was
then freeze dried in order to isolate the water soluble
polysaccharide fraction.

Preparation o f polysaccharide solutions
The •eze-dried exopolysaccharide was dissolved
in bidistilled water or in aqueous salt solution of the
desired concentration by prolonged gentle stirring
with a magnetic stirrer. For studies on the effect of
pH, test solutions were made by judicious adjustment
of the pH with a few drops of either 8 M HC1 or
NaOH solutions so as to maintain constant polysaccharide concentration.
Intrinsic viscosity
The viscosities of the polymer solutions were measured with a size 75 Cannon-Fenskesemi-micro capillary
viscometer (Cannon Instrument Co., State College, PA,
USA). The shear rate experienced by the polymer solutions in this viscometer was estimated to be in the range
of 175-670 s- 1. The effect of shear rate was examined by
the use of a narrower capillary (size 50, 110-450 s). The
difference in [~/] values was insignificant.
The viscometer was suspended in a thermostatically controlled water bath maintained at the required
temperature to within + 0.2°C, in the temperature
range 25-85°C. Equilibration time of 15-20 min was
allowed before measurements were made since the
setup was experimentally found to reach thermal
equilibrium within 10 min.
The relative viscosity of a given solution qr¢l (defined as the ratio between the solution viscosity and
the solvent viscosity q/qs) was determined by measurement of the relative efflux times in the capillary (Mays
and Hadjchristidis, 1991; Van Krevelen and Hoftyzer,
1976). Viscosity values were based on at least 2-3
efflux time readings taken for any given sample inserted into the viscometer. Variation between consecutive readings was lower than 1.5% and typically
around 0.5%. At least two independent viscosity determinations were performed for each concentration.
Four experimental points were used for extrapolation
to obtain the intrinsic viscosity [~/]. The intrinsic
viscosity is usually obtained from either the extrapolation of In ?]rel/C (Kraemer relation) or (?]rel- 1)/c
(Huggins relation) to zero polymer concentration, c.

However, (~/rel-- 1)/C ( -- rhp/C where ~/spis the specific viscosity rhp = (q - q,)/qs) of flexible polyelectrolyte solutions in pure water exhibits a unique dependence on concentration, i.e. it diverges rapidly
with dilution. This effect, due to polymer chain expansion, makes it extremely inaccurate to extrapolate
qsp/C to infinite dilution. Several equations have been
proposed in the literature to describe the concentration dependence of the viscosity of polyelectrolyte
solutions and to satisfactorily handle the extrapolation of the experimental data (Fuoss and Strauss,
1948; Fuoss, 1951; Liberti and Stivala, 1966; Yuan
et al., 1972). However, some questions have been
raised concerning the validity of these methods and
their ability to determine intrinsic viscosities with
precision (Yuan et al., 1972). Furthermore, it has been
shown that for flexible polyelectrolytes the apparent
divergence in rlsp/C as c ~ 0 is actually a maximum
followed by a decrease in qsp/C at very low c values.
The maximum is attributed to configurational changes due to dissolved gases and other impurities (Cohen and Priel, 1988). Consequently, in the present
study viscosity data were extrapolated to zero concentration by the combined standard Huggins and
Kraemer treatments. The choice of these latter
methods was also made because they appear to be
more widely used in the literature and thus facilitate
comparison of [q] obtained in the present study with
similar ones for other biopolyelectrolytes and with [q]
values for salt solutions which do not present any
extrapolation difficulty.

RESULTS

A typical set of data for the viscosity of PspP
solutions as a function of polymer concentration is
depicted in Fig. 1. The data for solutions in water and
in four different aqueous NaC1 solutions at 25 °C are
plotted in terms of rhp/C vs c. The reported (Cohen
and Priel, 1988; Yamanaka et al., 1990) divergence of
rhp/C as e -~ 0 in water is not observed in the concentration range examined here. The effects of salt concentration, type of counterions, and temperature on
the intrinsic viscosity of the PspP are summarized in
Table 1. The reported values are averages of at least
two independent determinations of [~/] with the variance always below 5% of the reported mean.
The intrinsic viscosity in NaCI solutions at 25°C is
plotted in Fig. 2 against I, the ionic strength of the salt
solution, I = (~ciZ~)/2p, where ci is the ion molar
concentration, Zi the number of charges on the ion
and p the solvent density. The plot shows an initial
sharp decrease of [~/] as the amount of salt is
increased at low NaC1 concentrations (I < 0.025),
followed by a very moderate decrease in [q] at higher
NaC1 concentrations.
The effect of the cation used to prepare the salt
solutions is shown in Fig. 3 (here [q] is plotted against
1 - ° 5 for reasons discussed in the following section).
As clearly observed in this figure the type of cation
and not only its valence are of importance. Sodium

Dilute solution viscosity of red microalga exopolysaccharide
50

.

.

.

.

t

.

.

.

.

i

.

.

.

0.5 M aqueous NaC1. F o r all test solutions in water,
there was a decrease in the [r/] values after the first
adjustment to acidic or basic pH. These lower Jr/]
values were very similar for all p H values. It can also
be seen from Fig. 4 that in the presence of external salt
the l-r/] of test solutions is unaffected by the p H
adjustments remaining essentially constant in the
2.0-11.0 p H range. This d e m o n s t r a t e s once again the
dimensional stabilization effect of salt solutions.

.

i1
~ 0 M NaCI

40
• _ei . . . . .

"0
v

....

DISCUSSION

~6~25e4M

Effect of ionic strength

~ ~1-.25e" M

30

--

v

0.00

1489

.-v ....

v

~-~'~if-

0.01

J-

0.03

0.02

Polysaccharide concentration (g/dl)
Fig. 1. Specific viscosity of Porphyridium sp. polymer solutions at 25°C. Solvents: (e) bidistilled water, (11)
6.25 x 10- 5 M, (&) 1.25 x 10 -4 M, (T) 0.01 M, and (4,) 0.1 M
NaC1 aqueous solutions. Each data point represents the
average of at least two independent determinations. The
lines represent the best fit linear regression.

salt is less effective in reducing molecular dimensions
than potassium. N o significant difference is observed
between intrinsic viscosity values for Ca ++ and
M g + +, the two divalent cations used in this study,
and at identical ionic strength [r/] is considerably
smaller for divalent cations than for m o n o v a l e n t ones.
Figure 4 shows the effect of p H on the [-~/] of P s p P
solutions p r e p a r e d in water and in the presence of

A decrease in intrinsic viscosity is observed when
the ionic strength increases (see Table 1). F o r a saltfree solution, electrostatic interactions due to the
charges on the polymer favour a stretched chain conformation as a result of long-range electrostatic effects. This behaviour results in higher intrinsic viscosity. Addition of a simple electrolyte screens these
intermolecular electrostatic interactions and causes
the polymer to assume a more flexible configuration
(wormlike chain), resulting in reduced intrinsic viscosity (Robinson et al., 1991; Smidsrod, 1970; Smidsrod
and Haug, 1971; Tinland and Rinaudo, 1989; Shatwell
et al., 1990; see Figs 1 and 3). Thus, recorded differences in viscosity between solutions of two different
batches of P s p P in water can stem from differences in
the a m o u n t s of salts present as impurities in the two
samples (due for example, to ineffective dialysis procedure), and need not necessarily be related to differences in molecular characteristics such as molecular
weight and its distribution or branching mode.

Effect of the nature of counterion
The fact that the different cations gave different [r/]
values (Table 1 and Fig. 3) suggests that the effect of
the counterion on P s p P is specific and d e p e n d e n t on
the type of counterion. The KC1 salt (compared to
NaC1) was more effective in reducing [r/] at very low

Table 1. Intrinsic viscosity [-~/](dl g - 1) of Porphyridium sp polysaccharide as a function of temperature and salt concentration
Salt concentration (M)
Temp.
(°C)
25

45
55
65
75
85

Salt type

0

6.25 × 10 -5

1.25× 10 -4

0.01

0.025

0.05

0.1

0.5

1.0

NaC1
CaC12
MgClz'6HzO
KC1

42.7

35.0
28.3
27.1
32.0

33.2
25.4
26.6
31.0

28.7
23.8
24.3
25.3

24.4

24.6

24.8
23.2
20.0
24.0

23.2
20.3
19.7
--

22.8
----

26.7
23.7
24.7
26.7
22.9
24.5
26.6
25.8
23.5

--------

--------

23.7
23.4
21.1
23.4
24.0
20.3
22.0
23.0
22.3

22.8
--22.3
--20.7
---

22.3
--22.5
--21.5
---

NaCI
CaC12
MgCI2 •6H20
NaC1
CaC12
NaC1
CaCIz
MgClz • 6HzO

36.0
35.5
36.5
34.6
32.4

Note: The [q] values given represent the average of at least two independent determinations.

1490

al.

et

E. ETESHOLA
45
I=0

A

4ol

40

i

r

-..°.°.

35

_

' '''"1

...

'

'

....

"1

'

'

' ' ....

I

#
"-..&
"---..•....

:35,
30

• -"-•-...•.


°°.

0
0
25

.9

30

C

i

2O
1 .e-5

i

q

i

i iiLil

~11

ILl

i

i

iJlll

1 .e-3

1 .e-4

1 .e-2

C
25

20

'

,

,

I

,

,

L

,

0.25

).00

I

i

~

,

~

I

0.50

1.00

0.75

Ionic Strength

(NaCI)

Fig. 2. Intrinsic viscosity of Porphyridiumsp. polysaccharide as a function of the ionic strength in aqueous
NaCI at 25°C. The insert is an enlargement of the low ionic strength range.

i

i

t

I

i

i

~

i

i

i

I

i

i

i

i

~

i

i

i

I

35

.

.

Ai,.,

•,D"

KCI .-'"

NaCI.-

30

.'"

.-

•.-

•-"

. •

.*'"
. . o.

0
0

._~

"I'X,

.o 25
°C~

..

CaCI 2
MgCI 2

C

° iiiiiii-

• • .........
°

20

ii-

. •°'•

I

I

I

I

v
i

1

i

[

i

10

I

I

L

I

I I I I

100

[ -0.S

Fig. 3. Intrinsic viscosity of Porphyridium sp. polymer for several monovalent and divalent cations. The
lines represent the best fit linear regression of the data.
and moderately low salt concentration ( < 1 × 10 -2 M).
The divalent cation Ca + + reduced Jr/] even further in
comparison with the monovalent cation Na +. As the
hydration level of K + is higher than that of Na + the
value of [q] is lower for the former because the hydrated ions of K + are more tightly b o u n d (Pasika,
1977). The divalent cations, which give even lower Jr/]
values, most probably form ion pairs to a much
greater extent than do the monovalent cations with
the carboxyl and the half ester sulfate groups of PspP.

Smidsrod (1970) suggested that Mg +÷ ions, whJc~
gave lower I-r/] than Na ÷ ions probably form ion pairs
with the carboxyl groups of alginate. Alternatively,
the greater effectiveness of the divalent cations may be
due to more effective shielding of the P. sp polyion
charges by bivalent ions (Schneider and Doty, 1954).

Determination of the flexibility parameter B
For many years it has been known that, for flexible
polyelectrolytes, intrinsic viscosity varies with 1-0.5

Dilute solution viscosity of red microalga exopolysaccharide
50

'

'

'

l

'

'

'

i

I

i

n

}

i

1491
i

i

i

I

i

i

n

I

160
rhamsan

150

140

40

¢

.__>,

[]

0
o

8

=o

>

xanthan

~

50
[]

.2

u

t-.

{

40

t_--

30

welan

~

30
P sp
~..~-,&

alginate

20

10

20

,
0

,

,

I

,

=

,

I

2

4

,

,

,

I

,

,

6

I
8

,

,

,

I
10

~

,

0

12

(with I the ionic strength), and that the slope of this
plot, S, is an indicator of chain flexibility (Shatwell
et al., 1990).
Smidsrod and Haug (1971) developed a simple
semi-empirical method based on this observation.
This is the so-called "B value method", in which the
parameter B is given by:
(1)

where [r/]o.1 is the intrinsic viscosity in 0.1 M NaC1,
and the persistence length a is:
a = 0.26/B

(2)

(for [r/l in dl g - 1 and a in nm). The constant (0.26) was
originally found by constructing a calibration plot for
a number of semi-flexible bioploymers. The persistence length is a measure of the length over which the
chain 'persists' in the direction of the first bond of the
chain. For stiff wormlike coils the Kuhn step length, l,
is given by (Anthonsen et al., 1993)
l = 2a.

l

i

l

I

i

~

i

5

i

[
10

~

i

i

I
15

! ..o.s

Fig. 4. Variation of the PspP intrinsic viscosity as a function
of the pH in water (0) and in 0.5 M NaC1 solution (11).

s/(l-r/]o.1) 1.3

h

0

,

pH

B =

K - carrageenan

(3)

Following this type of analysis, the intrinsic viscosity
data for PspP in NaC1 solutions are plotted in Fig. 5
as function of I - ° ' s . For comparison purposes data
for several other common polysaccharides are also
shown in the same figure. The linear dependence of
I-r/] on i - o . s in the salt concentration range of
0.01-1.0 M NaC1 is clearly observed. Both qualitatively and quantitatively the behavior of PspP is in

Fig. 5. Variation of intrinisc viscosity with ionic strength, I,
for Porphyridium sp. polysacharide (O), rhamsan (V), xanthan (D), welan (i), alginate (A), and x-carrageenan (O).
Data for alginate, welan and rhamsan were taken from
Robinson et al. (1991), for x-carageenan from Slootmaekers
et al. (1988) and for xanthan from Shatwell et al. (1990):

accord with that observed for other polysaccharides.
A linear dependence of [r/] on 1-0.5 is also observed
for all other salts examined here (Fig. 3).
At high ionic strength, charge screening reduces the
influence of coulombic interactions on polymer conformation, whereas at low ionic strength these interactions become important and do affect conformation.
This fact may be deduced from the nonlinearity observed in the plot in Fig. 6, which includes data on the
effect of NaC1 at very low ionic strength and over
a considerably broader concentration range than that
shown in Fig. 5. The decrease in [~/] with increasing
ionic strength implies the existence of attractive interactions between chain elements, possibly of electrostatic origin. The slope change observed in the plot
around I ~ 0.01 M may hint at a transition in the
P. sp polyion conformation, most likely reflecting
a contraction and conformational ordering (with increasing ionic strength) of the polyion chain from
a highly stretched conformation to a stiff but nevertheless wormlike chain. Similar transitions have been
reported in the literature (Holzwarth, 1981; Slootmaekers et al., 1988).
We will examine the PspP Jr/] data within the
concentration range 0.01-1.0 M NaCI in greater detail
(Fig. 5). Applying eqs (1)-(3) to the data presented in
Fig. 5, we obtain B = 0.0074 for the stiffness parameter,

1492

E. ETESHOLAet al.
45

. . . .

I I , l l l l l l l l l l l l l l l l ,

I

1=0

40

~'~ 35
u0

.~_
°~-

30

25

20

~ , , 1 ~
0

, I , ~ , 1 , ~

25

50

75

, ,ll
100

,~

,I
125

I -.0.5

Fig. 6. The dependence of the intrinsic viscosity of Porphyridium sp. polymer on the concentration of added sodium
chloride at 25°C.

a = 35 nm for the persistence length and 70 nm for the
K u h n step length for PspP. These values are reported
in Table 2 in comparison with other biopolyelectrolytes. The value of 0.0074 estimated for B, which is
a measure of chain flexibility (higher B value corresponds to a more flexible chain), is close to values

obtained for rigid helices such as xanthan and D N A ,
but less than those obtained (Smidsr~d and Haug,
1971) for carboxylated polysaccharides with characteristically more flexible backbones, such as alginate
and carboxymethyl cellulose (Axelos and Thibault,
1991; Slotmaekers et al., 1988; see Table 2). Therefore
the modest change in [r/] as a function of ionic
strength (Fig. 5) may be taken to attest to the stiffness
of the P s p P chain structure. A possible explanation
for this property is that the P. sp biopolymer has
a configuration which maintains the chain molecule in
a stiff form similar to the situation reported for D N A
(Cox, 1960). We note that the question of whether the
P s p P molecule is a single or a multiple stranded helix
in aqueous solution may not have been answered by
the experiments reported here. However, preliminary
X-ray diffraction studies (Eteshola et al., submitted)
indicate the possibility of a two-fold helical structure.
In terms of property-structure relationships, the low
expansion coefficient observed for the P s p P chain (the
linearity observed in Fig. 5) could also be due to
a complex branching mode (Painter, 1983; Flaibani
et al., 1989). Yuan et al. (1972) have reported that
branching can curtail the ability of a polyion to
expand.
The persistence length was estimated as already
indicated above. In addition, the K u h n step length
was estimated (data not shown) by interpolation from
Figure 10 in the paper by Smidsrod and Christensen
(1991), which mentions an empirical correlation between B and the K u h n length. This relationship also
reveals that the P s p P has a chain stiffness comparable
to that of D N A and xanthan. Thus, from the various
stiffness indices reported in Table 2, it may be concluded that the overall molecular conformation and
dimensions of the P. sp biopolymer chains as reflected
by the intrinsic viscosity bear some similarities to

Table 2. Stiffness indices for several biopolyelectrolytes
Polyelectrolyte
Porphyridium sp.

Rhamsan*
Xanthan
Xanthan*
Xanthan ps. PXO61~
Xanthan ps. 556 ~
DNA*
Pectin (C72)~
Alginates-guluronate-rich *
-mannuronate-rich*
x-carrageenan~
Carboxymethyl cellulose*
* Robinson et al., 1991.
t Tinland and Rinaudo, 1989.
Slootmaekers et al., 1988.
~Shatwell et al., 1990.
~Axelos and Thibault, 1991.
** By viscometry.
tt By light scattering.

B
0.0074
0.003
0.00525 t

0.0055
0.017
0.031
0.040
0.065

a (nm)

l (nm)

35.1
88
50*
7"* (coil)
40** (helix)
42.2
39.2
45
15.3
7.8
6.5
6.8 t*
3.7**
4.1

70.2
-100
84.4
78.4
90
30.6
1
---

Dilute solution viscosity of red microalga exopolysaccharide
those of D N A and xanthan. F r o m the above results
and observations, it is hypothesized that the P. sp
polysaccharide chain molecules adopt relatively stiff
conformation in dilute solutions over a relatively wide
salt concentration range.
Effect of p H and ionic strength
The intrinsic viscosity decrease as a result of pH
changes (Fig. 4) can be regarded at most as moderate,
and some of the decrease, especially under acidic
conditions, is negligible. The P s p P seems to be somewhat more stable under acidic than basic conditions.
The dependence of the intrinsic viscosity of P s p P
solutions on the p H in the presence of external salt is
characteristic for a polyelectrolyte solution. The large
conformational changes due to strong screening of
coulombic interactions by the salt masks the small
changes due to the p H effects discussed above. Similar
observations have been made for the polyelectrolytes
Na-xanthan and Na-carboxymethyl-cellulose (Rinaudo
and Milas, 1978).
CONCLUSION
This paper represents the first attempt to study
chain flexibility and conformational features for Porphyridium sp. polysaccharide in dilute aqueous and
salt solutions by intrinsic viscometry.
Polyelectrolytic behaviour is confirmed by the decrease of I-q] with the addition of neutral salt. C o m bined data for very low to moderate ionic strength
indicate a change in slope in the plot of E~/] vs 1 - ° 5 ,
which may hint at a transition in P s p P chain confirmation from a highly stretched to a stiff, wormlike
chain. However, no clear o r d e ~ d i s o r d e r conformational transition as a result of ionic strength changes
was detected. The stiffness parameter was deduced
from the dependence of [q] on ionic strength; from the
value obtained it is concluded that the stiffness of
P s p P chains is in the same range as xanthan and
D N A . Due to the stiff configuration adopted by the
chain molecules in solution, a relatively low sensitivity
to increasing ionic strength is observed. The low thermal expansion coefficient displayed may be related to
the detailed primary and chemical structure of the
polymer. The polymer showed specificity to counterion type and valency.
Acknowledgement--MG acknowledges the support of the
Israel Science Foundation administered by the Israel Academy of Sciences and Humanities.
REFERENCES

Anthonsen, M. W., Varum, K. J. and Smidsrod, O., 1993,
Solution properties of chitosan: conformation and chain
stiffness of chitosan with different degree of N-acetylation. Carbohydr. Polym. 22, 193-201.
Arad (Malis), S., Friedman, O. D. and Rotem, A., 1988, Effect
of nitrogen on polysaccharide production in a Porphyridium sp. Appl. Envirom. M icrobiol. 54, 2411-2414.
Axelos, M. A. V. and Thibault, J.-F., 1991, Influence of the
substituents of the carhoxyl groups and of the rhamnose

1493

content on the solution properties and flexibility of pectins. Int. J. Biol. Macromol. 13, 77 82.
Cohen, J. and Priel, Z., 1988, Viscosity of dilute polyelectrolyte solution. J. chem. Phys. 88 (11), 7111-7116.
Cox, R. A., 1960, The influence of ionic strength on the
viscosity of ribonucleic acid and other polyelectrolytes.
J. Polym. Sci. 47, 441-447.
Eteshola, E., Rideout, M. J., Brownsey, G. J., Cairns, P.,
Gottlieb, M., Morris, V. J. and Arad (Malis), S., submitted, Study of the rheology and morphology of aqueous
preparations from red microalga exopolysaccharide.
Carbohydr. Polym.
Flaibani, A., Olsen, Y. and Painter, T. J., 1989, Polysaccharides in desert reclamation: composition of exocellular
proteoglycan complexes produced by filamentous
blue-green and unicellular green edaphic algae. Carbohydr. Res. 190, 235-248.
Fuoss, R. M., 1951, Polyelectrolytes. Disc. Farad. Soc. 11,
125-134.
Fuoss, R. M. and Strauss, U. P., 1948, Polyelectrolytes.
II. Poly-4-vinylpyridonium chloride and poly-4-vinylN-n-butylpyridonium bromide. J. Polym. Sci. 3(2),
246-263.
Geresh, S. and Arad (Malis), S., 1991, The extracellular
polysaccharide of red microalgae: chemistry and rheology. Bioresource Technol. 38, 195 201.
Geresh, S., Lupescu, N. and Arad (Malis), S., 1992, Fractionation and partial characterization of the sulphated
polysaccharide of Porphyridium. Phytochemistry 31(12),
4181-4185.
Heaney-Kieras, J., Roden, L. and Chapman, D. J., 1977, The
covalent linkage of protein to carbohydrate in the extracellular protein~olysaccharide from the red alga Porphyridium cruentum. Biochem. J. 165, 1-9.
Heaney-Kieras, J. and Chapman, D. J., 1976, Structural
studies on the extracellular polysaccharide of the red alga
Porphyridium cruentum. Carbohydr. Res. 52, 169-177.
Heaney-Kieras, J., Kieras J. F. and Bowen, D. V., 1976,
2-O-Methyl-D-glucuronicacid, a new hexuronic acid of
biological origin. Biochem. J. 155, 181 185.
Holzwarth, G. M., 1981, Is xanthan a wormlike chain or
a rigid rod? in Solution Properties of Polysaccharides
(Edited by D. A. Brant), pp. 15-23. ACS Symp. Ser. 150,
American Chemical Society, Washington, DC.
Jones, R. E., Speer, H. L. and Kury, W., 1963, Studies on the
growth of the red alga Porphyridium cruentum. Physiol.
Plant. 16, 636-643.
Jones, R. F., 1962, Extracellular mucilage of the red alga
Porphyridium cruentum. J. Cell. Comp. Physiol. 60, 61-64.
Van Krevelen, D. W. and Hoftyzer, P. J., 1976, Properties of
Polymers--Their Estimation and Correlation with Chemical Structure. Elsevier, Amsterdam.
Lee, E. J. and Chandrasekaran, R., 1992, The "pseudo
double-helical" structure of the gel-forming capsular
polysaccharide from Rhizobium trifolii. Carbohydr. Res.
231, 171-183.
Liberti, P. A. and Stivala, S. S., 1966, On the viscosity of
aqueous polyelectrolytes. Polym. Lett. 4, 137-145.
Mays, J. W. and Hadjchristidis, N., 1991, Polymer characterization using dilute solution viscometry, in Methods of
Polymer Characterization (Edited by H. G. Barth and
J. W. Mays), pp. 227 269. Wiley, New York.
Medcalf, D. G., Scott, J. R., Brannon, J. H., Hemerick, G. A.,
Cunningham, R. L., Chessen, J. H. and Shah, J., 1975,
Some structural features and viscometric properties of
the extracellular polysaccharide from Porphyridium
cruentum. Carbohydr. Res. 44, 87-96.
Painter, T. J., 1983, Algal polysaccharides, in The Polysaccharides, Vol. 2 (Edited by G. O. Aspinall), pp. 195 285.
Academic Press, Inc., NY.
Pasika, W. M., 1977, Polysaccharide polyelectrolytes, in
Extracellular Microbial Polysaccharides (Edited by P. A.
Sandford and A. Laskin), pp. 128-143. ACS Symp. Ser.
45, Washington, D.C.

1494

E. ETESHOLAet al.

Percival, E. and Foyle, R. A. J., 1979, The extracellular
polysaccharides of Porphyridium cruentum and P.
aerugineum. Carbohydr. Res. 72, 165-176.
Ramus, J., 1973, Cell surface polysaccharides of the red alga
Porphyridium in Biogenesis of Plant Cell Wall Polysaccharides (Edited by F. Loewus), pp. 333-358. Academic
Press, New York.
Ramus, J., 1986, Rhodophyte unicells: biopolymer, physiology and production, in Algal Biomass Technologies--An
Interdisciplinary Perspective (Edited by W. M. Barclay
and R. P. McIntosh), pp. 51-55. J. Cramer, Berlin.
Ramus, J., Kenney, B. E. and Shaughnessy, E. J., 1989, Drag
reducing properties of microalgal exopolymers. Biotechnol. Bioengng 33, 551~557.
Rinaudo, M. and Milas, M., 1978, Polyelectrolyte behavior
of a bacterial polysaccharide from Xanthomonas
campestris: comparison with carboxymethylcellulose. Biopolymers 17, 2663 2678.
Robinson, G., Manning, C. E. and Morris, E. R., 1991,
conformational and physical properties of the bacterial
polysaccharides Gellon, Welan and Rhamsan in Food
Polymers, Gels and Colloids (Edited by E. Dickson), pp.
22-33. The Royal Society of Chemistry, Cambridge, U.K.
Savin, J. G., 1978, Oil recovery process employing thickened
aqueous driving fluid. Mobil Oil Coporatio, N. Y., U. S.
Pat. 4 079 544.
Schneider, N. S. and Doty, P., 1954, Macro-ions IV. The ionic
strength dependence of the molecular properties of sodium
carboxylmethyl cellulose. J. phys. Chem. 58, 76~769.
Shatwell, K. P., Sutherland, I. W. and Ross-Murphy, S. B.,
1990, Influence of acetyl and pyruvate substituents on the
solution properties of xanthan polysaccharide. Int. J.
Biol. Macromol. 12, 71-78.

Simon, B., Geresh, S. and Arad (Malis), S., 1992,
Degradation of the cell-wall polysaccharide of Porphyridium sp. (Rhodophyta) by means of enzymatic activity of its predator, Gymnodium sp. (Pyrrophyta). J.
Phycol. 28, 460-465.
Slootmaekers, D., De Jonghe, C., Reynaers, H., Varkevisser,
F. A. and Bloys, van Treslong, C. J., 1988, Static light
scattering from k-carrageenan solutions. Int. J. Biol.
Macromol. 10, 16(~168.
Smidsrod, O., 1970, Conformation of polyelectrolytes in
solution. Carbohydr. Res. 13, 359.
Smidsrod, O. and Christensen, B. E., 1991, Molecular structure and physical behaviour of Seaweed colloids as
compared with microbial polysaccharides, in Seaweed
Resources in Europe: Uses and Potential (Edited by
M. D. Guiry and G. Blunden), pp. 185-217. Wiley,
New York.
Smidsrod, O. and Haug, A., 1971, Estimation of the relative
stiffness of the molecular chain in polyelectrolytes from
measurements of viscosity at different ionic strengths.
Biopolymers 10, 1213 1227.
Tinland, B. and Rinaudo, M., 1989, Dependence of the
stiffness of the xanthan chain on the external salt concentration. Macromolecules 22, 1863 1865.
Yamanaka, J., Matsuoka, H., Kitano, H., Hasgawa, M.
and Ise, N., 1990, Revisit to the intrinsic viscosity
molecular weight relationship of ionic polymers. 2.
Viscosity behavior of salt-free aqueous solution of
sodium poly(styrenesulfonates). J. Am. Chem. Soc. 112,
587 592.
Yuan, L., Dougherty, T. J. and Stivala, S. S., 1972, A semiempirical approach to the viscosities of polyelectrolyte
solutions. J. Polym. Sci. A2, 10, 171-189.