British Phycological Journal

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The polysaccharides of green, red and brown

seaweeds: Their basic structure, biosynthesis and
function

Elizabeth Percival

To cite this article: Elizabeth Percival (1979) The polysaccharides of green, red and brown
seaweeds: Their basic structure, biosynthesis and function, British Phycological Journal, 14:2,
103-117, DOI: 10.1080/00071617900650121

To link to this article: http://dx.doi.org/10.1080/00071617900650121

Published online: 17 Feb 2007.

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 Br. phycol. J. 14:103-117
1 June 1979

THE POLYSACCHARIDES OF GREEN, RED

AND BROWN SEAWEEDS: THEIR BASIC
STRUCTURE, BIOSYNTHESIS AND
FUNCTION*
By ELIZABETH PERCIVAL
The Bourne Laboratory, Royal Holloway College, Egham Hill, Egham, Surrey, TW20 OEX

This is a very ambitious title since there is comparatively little known of the
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biosynthesis of these polysaccharides and not a great deal has been proved of
their function. It is known that they fulfil a variety of functions and show a
corresponding range of structures. They have as constituents the various neutral
sugars and sugar acids which are found in land plants, but some also contain
acidic half ester sulphate groups attached to hydroxyl groups of the sugars.
These are not found in other plants but occur in animal polysaccharides.
Although the hexose sugars glucose, galactose and mannose, which are present
in many of these polysaccharides, have an identical chemical formula their
constituent atoms have different special arrangements and this together with
the way they are linked permits the elaboration of a vast number of poly-
saccharides with different shapes and in consequence different properties. In
addition tile pentoses, xylose and arabinose, and the deoxyhexoses, rhamnose
and fucose allow a further range of structures, and the uronic acids, glucuronic,
mannuronic and guluronic and the half ester sulphate groups introduce the
possibility of different combination with metal ions, as well as different shaped
macromolecules or in current terminology conformations.
Thus the very many different polysaccharides which have been found in
seaweeds are only a small proportion of the possible structures, and we must
assume that in the course of evolution they have proved to have certain
advantages in the environments where the seaweeds are found. Some are ahnost
certainly food reserves and others are constituents of cell walls, but many
have functions which at present are not at all well understood.
The majority of the chemical studies have been carried out on extracts of
mixtures of whole weeds in different stages of growth. In many cases the chemi-
cal structures are only the average of all the molecules present in a particular
type of polysaccharide which is often a family of polydisperse heteromolecules,
all built up on the same general plan, but which differ in their fine details. It
appears that modifications in the structure of the macromolecule are brought
about by the plant to enable the polysaccharide to fulfil a different function.
Some are linked to protein and are therefore strictly speaking proteoglycans.
Nevertheless the chemical studies and the fine details of the chemical constitu-
tions of the more complex of these polysaccharides will be described only
* Presidential Address presented to the British Phycological Society on 3 January 1979.
103
0007-1617/79/020103-1-15 $02.00/0 (~ 1979 British Phycological Society
A
 104 E. PERCIVAL

in so far as they affect the physical properties and hence their function in the
seaweed.

CHLOROPHYCEAE
During the past 25 years my colleagues and I have examined the carbohydrates
of nine different genera of green seaweed and some twenty different species.

FOOD RESERVE
The majority metabolize a small proportion of starch which closely resembles
land plant starches, although the starch granules are smaller and less structurally
defined and the molecules of anaylose and amylopectin appear to have a smaller
molecular weight (LOve et al., 1963). This clearly serves as a food reserve
polysaccharide. A small amount of work on the biosynthetic pathway has been
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carried out by the Japanese (Kashiwabara, Suzuki & Nisizawa, 1965). They
isolated an amylase from Cladophora and from Ulva and a phosphorylase was
isolated from U. pertusa which transfers ~-D-glucosyl residues from glucose 1P.

HO//~ O\ OH b3SO~ H20H 0~o H

OH OSO3 ON

&-L-Rhamnose2- sulphate B-D-Galactose

4 -sulphate
FIG. 1.
TABLE I.

Ulva lactuca
Acrosiphonia centralis
Entromorpha compressa
Urospora penicilliformis
Urospora wormskioldii
Codiolum pusillum
l
/
Sulphated glucuronoxylorhamnans
Negative [C~]D
J
Aeetabularia crenulata Sulphated glucuronoxylorhamnogalact an
Negative [aJD
Chaetomorpha eapillaris
Chaetomorpha linum [
Cladophora rupestri~ }- Sulphated xyloarabinogalactans
Caalerpa filiformis 1 Positive [~]D
Codium fragile J
 Polysaccharides of green, red and brown seaweeds 105

In contrast the Dasycladales, e.g. Acetabularia species, have a little starch in

the chloroplasts but none has been detected in the cytoplasm. Instead a fructan
of the inulin-type (Bourne, Percival & Smestad, 1972) appears to replace the
starch as the active metabolite, and UDPG-phosphorylase has been shown to
play a part in the polysaccharide synthesis.
Low molecular weight fl-l,3-1inked glucans have been found in Codiolum
pusillum (Carlberg & Percival, 1977) and in Caulerpa simpliciuscula (Hawor-
thorne, Sawyer & Grant, 1979). The latter has 1-2 small branches linked
through C-6. It appears to be associated with a soluble starch of the same mole-
cular size in the ratio of 3 molecules of fl-glucan per molecule of e-linked
glucan. The authors suggest that the fi-glucan is a specific reserve for the syn-
thesis of cell wall which is laid down during the final stages of wound healing.

SULPHATED POLYSACCHARIDES
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The major polysaccharides, however, in all the species examined are poly-
disperse heteropolysaccharides in which at least some of the hydroxyl groups
of the sugar residues are substituted by half ester sulphate groups (Fig. 1).
These are very strong acids and exist in vivo as salts of various metals.

TABLE II. Cladophorales--Motecular proportions of the sugars in the water soluble poly-
saccharides

Galactose Arabinose Xylose

Cladophora
rttpestris 1"0 l 1 0"4
#laucescens 1"0 1"3 08
albida 10 1"8 09
I 1 "0 0"7 0"4
I[ 1 "0 09 05
glomerate F.W. 1-0 09 07
Rhizoclonium
implexum l '0 0'6 0'45
riparum 1"0 O"8 0"4
sp. F.W.
Chaetomorpha
linum 1"0 2"5 0"7
aerea 1"0 12 0"45
eapillaris 1"0 2"6 0"7

The different genera of green seaweeds fall into several main groups according
to the type of polysaccharide produced (Table I). Structural studies on all of
these and similar polysaccharides has established that the proportion of the
sugars vary from genus to genus and even among species of the same genus.
Table II shows the different proportions of the constituent sugars in species of
the Cladophorales. The modes of linkage and the points of attachment of the
sulphate groups have been established for the majority of these polysaccharides
(Percival, 1978a). They are all extremely complex highly branched molecules
which do not appear to have a backbone or simple repeating unit. N o r do they
appear to have long chains of a single sugar since evidence of the mutual linkage
 106 E. PERCIVAL

of the different sugars has been obtained for the different groups. Even after
purification extracts still contain a small proportion of protein and they are
probably proteoglycans.
No conclusive biosynthetic studies have been carried out on these poly-
saccharides. All attempts to isolate active enzymes from the seaweeds have
proved unsuccessful.

Functions
A variety of functions have been suggested for these polysaccharides. In
vivo they exist as mucilages or gels of varying stiffness. This varies with the
particular metals with which they are associated. One suggestion is that as
anionic electrolytes they maintain the ionic equilibrium in the cell. Often they
are present as a stiff gel outside the ceil. There they serve to repel the attack of
other organisms, and at the same time strengthen the cell wall and give
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flexibility to the plant to enable it to withstand wave action. A further role of

these polysaccharides may be to prevent desiccation in plants exposed by the
tides. They are hygroscopic materials which have the property of retaining
moisture under dry conditions.
TABLE III.

Carbohydrate extracted
Extraction media % dry wt of weed

Distilled water 18"2

After removal of Ca + + followed by distilled water 19"4
Natural seawater 4"7
Synthetic mixture (pH 8) containing H3BOs (0"024 g 1-~)
CaCI22H20 (0"273 g l-a) 4.5

Haug (1976) carried out experiments with live fronds of Ulva lactuca. He
showed that if such fronds are placed in distilled water the glucuronoxylo-
rhamnan (the sulphated polysaccharide) loses its gelatinous character and passes
out into the water. He then experimented with different salts present in sea-
water and added these in turn to the distilled water and found only if borate
and calcium together in the concentration and p H found in seawater were
added to the distilled water did the polysaccharide remain in the plant (Table
III). Haug postulated that the borate complexed with the free hydroxyl groups
on C-2 and C-3 of the 1,4-1inked rhamnose units in this polysaccharide as shown
in Fig. 2, and that the Ca ++ stabilized the borate complex, and that these
structures maintained the polysaccharide in the stiff gel state. Structural studies
(Percival & Wold, 1963) had shown that sulphate groups were linked to some of
the hydroxyl groups at C-2 of the rhamnose units. The presence of sulphate on
C-2 of the rhamnose would prevent these units complexing with borate. By
adding or removing these sulphate groups Ulva is thus able to regulate the
stiffness of the gel of its polysaccharide while growing under natural conditions.
In the same way the presence of calcium ions alone causes a dilute solution of
the sulphated polysaccharide extracted from Cladophora rupestris to form a
stiff gel (Percival, unpublished work).
 Polysaccharides of green, red and brown seaweeds 107

STRUCTURAL POLYSACCHARIDES
While the sulphated polysaccharides may comprise part of the cell walls
many of the Chlorophyceae also synthesize comparatively simple xylans and
mannans. The xylans contain chains of ~-l,3-1inked xylose units (Mackie &
Percival, 1959) which X-ray studies have shown form an open wire spring-like
helix with three chains forming a triple helix (Atkins & Parker, 1969) thus
giving a strong fibrous structure which constitutes the main skeletal poly-
saccharide of such species as Caulerpafil~formis. In other green algae mannans
appear to be the structural polymer. Codium fragile synthesizes a/M,4-1inked
mannan (Love & Percival, 1964) which serves as a partially crystalline skeletal
component (Frei & Preston, 1968). The Codiolum stage in the life history of
Urospora wormskioldii also contains a fl-l,4-1inked mannan (Carlberg &
Percival, 1977). The prostrate form of this alga is practically devoid of mannan,
supporting the view that this is a structural polysaccharide since both forms of
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Urospora contain a small proportion of cellulose. It is possible that the mannan

is laid down with this, as in the hemicelluloses of land plants to give added
strength to the Codiolum which is the erect stage of this alga.

¢[- L - R h a m n o s e 2 sulphate

Y..

9
1

FIG. 2.

RHODOPHYCEAE
FOOD RESERVE

The Rhodophyceae also metabolize a starch, known as floridean starch, as

their food reserve material (Nagashima et al., 1969). It differs from the Chloro-
phycean starch in that it is devoid of amylose (Peat, Turvey & Evans, 1959).
It gives a red violet colour with iodine, similar to that given by glycogen. It is
attacked by all the enzymes which hydrolyze starch. Floridean starch differs
from land plant amylopectins in having average chain lengths of about 11-13
compared with 25 for land plant starches. To summarize, in many of its pro-
perties Rhodophycean starch resembles the amylopectin of the higher plants,
 108 E. PERCIVAL

but in others it is intermediate between the latter and the animal polysaccharide,
glycogen. Its granules are again less organized than the grains of land plant
starches and they are hydrolyzed more readily. Japanese workers (Nagashima,
Nisizawa & Hori, 1971) have shown that ADI4P glucose was utilized by an
c~-l,4-glucosyltransferase in the red seaweed Serratieardia maxima with floridean
starch as the primer. /%Amylolysis of the product released maltose with ~4C
labelled glucose. These workers showed that the starch granules were localized
exclusively outside the chloroplasts and that most of the synthesis occurred
outside.
In Rhodymenia palmata (dulse) the major polysaccharide is a water-soluble
xylan made up of 1,3- and 1,4-1inked units in a purely random sequence
(Bjorndahl et al., 1965). No evidence for floridean starch in this seaweed has
been advanced and the solubility of the xylan in water suggests that it is a food
reserve polysaccharide. More recent work has shown that a second fraction
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comprising only 1,4-linkages can be extracted with dilute alkali (Turvey &
Williams, 1970), so that it is possible that it also serves as a structural polymer.

A B A B
CH20H H£) , CH20H C.HzOH
HO .J 0 HO J - - - ~ O ]'-mO,

/ OH OH
L-Galactose D- Galactose
i C.3
C--0
HOOC/" b I
I C,H2 0

OH "OH 0S07 - " .

OH
D-galactose 6-sulphote D-golactose2,6-disulphate Galactose4, 6-0 - I- carboxylethylidene

HO~--~OH H O ~ O H
OH
a -I, 4-1inked 3 6-anhydm-L-galactose 6-O-MethyI-D-goloctose
FIG. 3.

Other 1,3- and mixed 1,3-; 1,4-1inked xylans have been separated from
Porphyra umbilicalis, Laurencia pinnatafida (Turvey & Williams, 1970) and
Chaetangiumfastigiatum (Cerezo, 1972) all of which also synthesize a galactan.
Frei & Preston (1964) mechanically separated the cell wall of P. umbilicalis
and Bangiafuscopurpurea and showed that they consisted of 1,3-1inked xylans
in a random network of microfibrils. Both these weeds synthesize fl-l,4-1inked
mannans which exist in a cuticle outside the xylan. An enzymic extract from
P. umbilicalis was shown to have mannanase activity on the/3-1,4-1inked mannan
of ivory nut.
 Polysaccharides of green, red and brown seaweeds 109

SULPHATED POLYSACCHAR1DES
Quantitatively the major polysaccharides of the Rhodophyceae are galactans
consisting entirely of galactose or modified galactose units. They are known
commercially as agar, carrageenan and Danish agar. The major difference
between the agars and carrageenans is that the former contains D- and L-
galactose units whereas the latter consists entirely of the D-sugar. Besides
being substituted by half ester sulphate groups some of the galactose units in
all these polysaccharides are present as 3,6-anhydrogalactose (Fig. 3). They are
present in vivo as mucilages and gels. The proportions and site of the sulphate
groups and the amount of 3,6-anhydrogalactose differ in the polysaccharides
from different genera. The overall shape of the macromolecules, which comprise
twofold helices, determines the physical properties and these in turn are affected
by the amount and point of attachment of the sulphate groups and also the
proportion of 3,6-anhydrogalactose present (Percival, 1978b). The higher the
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latter is the better are the gelling properties of the polysaccharide. It is worth
pointing out that recent work on the sporophytic and gametophytic forms of
Chondrus cri~pus (McCandless, Craigie & Walter, 1973) and on Gigartina spp.
(Pickmere, Parsons & B~iley, 1973; McCandless, Craigie & Hansen, 1975) has
shown different structural carrageenans in the two life forms of these seaweeds.

HO~H2OSO30,
H ~o >H
OH OH

G a l a c t o s e - 6 - sulphate 3 6 - anhyd rogalactose

FIG. 4.

Biosynthesis
Enzyme preparations isolated from PotThyra umbilicalis (Rees, 1961) and
from Gigartina stetlata (Lawson & Rees, 1970) converted L-galactose 6-sulphate
and D-galactose 6-sulphate respectively into the corresponding 3,6-anhydro-
galactose (Fig. 4). Biologically this results in the removal of a "kink" in the
polysaccharide chain and permits more extensive double helix formation to give
a more compact rigid gel framework. This adaptation allows the plant to produce
stiffer gels when, for example, it is exposed to more severe wave action. Little
is known of the biosynthesis of the macromolecule of these galactans. Radio-
active studies with 14C indicated that uridine diphosphate glucose is converted
into UDP-galactose and this is utilized in the synthesis of these galactans.
 t 10 E. P E R C I V A L

Functions
The functions of these polysaccharides in the seaweed are probably very
similar to those of the sulphated polysaccharides of the Chlorophyceae. Working
with Ceramium the Leeds workers (Chamberlain & Evans, 1973) consider,
that after spore release, the galactan released with the spores acts as a protective
mucilaginous layer for the spores.

PHAEOPHYCEAE
LAMINARAN
In the Phaeophyceae the food reserve materials are a sugar alcohol, mannitol
and a fl-l,3-1inked g!ucan laminaran. This is a water-soluble polysaccharide
containing approximately 20-25 glucose units. In some species there are two
types of chains, G-chains which are terminated at the reducing end with glucose
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and M-chains which are terminated by mannitol (Percival, 1978c) (Fig. 5). In
CH2OH r CHzOH "I
0 0

uN L OH "In HOICH
HCOH
f
HCOH
CHzOH

M - Chains

C.H2 ~ r

Oo
O
CHzOH 1 CHzOH

N ,OH

(3- Chafns
FIG. 5.

addition the laminaran, for example, from Eisenia bicyclis, contains as much
as a third of 1,6-1inked units in the chains, occasionally the chains may be
branched at C-6. Bidwell (1967) has shown that the mannitol and laminaran
are active metabolites which can be interconverted. Quatrano & Stevens (1976)
have shown that laminaran isolated from the cytoplasm of developing zygotes
of Fucus spp. decreased during the first 7 h of wall assembly while cellulose in
the wall increased and this occurred with no external source of carbon or light.
An exo-fl-l,3-glucanase released from a particulate fraction of the zygotes
with Triton X hydrolyzed native laminaran to glucose.

ALGINIC ACID
Quantitatively the major polysaccharide of the brown seaweeds is alginic
acid. It is a polysaccharide consisting of unbranched chains comprising blocks
 Polysaccharides of green, red and brown seaweeds 111

of contiguous fl-l,4-1inked D-mannuronic acid and blocks of contiguous

e-l,4-1inked L-guluronic acid (Fig. 6), the average lengths of the blocks being
about 20 units and these are interspersed with a statistical mixture of the two
acids (Haug et al., 1967). The proportions of the two acids have been found to
vary in the extracts from different species and from different parts of the same
weed (Haug, Larsen & Smidsrod, 1974).
COOH

o O .... OOH
o O . . . .

,., I \ OH HO J l .... o_j ~ OH HO /" I

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13- D - M a n n u r o n i c acid d,- L - Guluronic a c i d

FIG. 6.

Alginate is present in the plant as salts of different metals, primarily sodium

and calcium. It occurs in both the intercellular region and the cell walls and it is
considered that its biological functions are principally of a structural and ion
exchange type. Alginic acid enriched in polymannuronic acid characterizes
young cell wall tissue and/or intercellular regions, whereas polyguluronic-rich
alginic acid appears to be located in the cell wall proper (Haug, Larsen &
Baardseth, 1969). This is to be expected since it is known that guluronic acid rich
alginates have a high affinity for Ca++, and it is these ions which are mainly
responsible for gel strength. Rees (1972) has shown how the calcium can form
with the G-blocks of an alginate what he calls the "egg box" structure in which
the Ca ions fit into the folds of the guluronic acid chains (Fig. 7). The Ca ++

FIG. 7. Egg box diagram of L-guluronic acid calcium.

ions are co-ordinated with the oxygen atoms in neighbouring chains thus
stabilizing the whole structure and being firmly held. This results in stiff gels
necessary for a semirigid cell wall. X-ray evidence and conformational analysis
(Whittington, 1971) also indicates the polymannuronic acid may be more
 112 E. P E R C I V A L

flexible than polyguluronate in the sense that rotations can take place about the
glycosidic bonds in the former, whereas the units appear to be more rigidly
held in twofold helices in the latter.

Biosynthesis
Japanese work (Abe et al., 1973) on the brown seaweed lshige okamurai
indicates that alginate is polymerized in the cytoplasm and then transported to
the cell surface. Lin & Hassid (1966) separated a GDP-D-mannuronic acid from
the brown seaweed, Fueus gardneri and considered that it was the precursor of
polymannuronic acid in young tissue. They also tentatively identified a trace
quantity of a GDP-guluronic acid. However, in more recent work the Nor-
wegians (Madgwick; Haug & Larsen, 1973a) have extracted an epimerase from
Pelvetia canaliculata and showed that this converted polymannuronic acid into
a mixed poly-D-mannuronic-L-guluronic polymer. They were able to monitor
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this change by tritium incorporation.

COOH COOH COOH . .. .
v~ ./n

0 0 0 0. . . .

.... 0

I Lyase

COOH COOH COOH COOH

0 0 -0 OH

F~G. 8. A chain of 1,4-1inkedD-mannuronic acid cleared by enzyme.

It has been shown (Quatrano & Stevens, 1976) that cell walls of zygotes of
several species of Fucus 4 h after fertilization consist of 60 ~ alginate, 20
cellulose and 20 ~ fucan and that these proportions persist although additional
polysaccharides are synthesized. The alginate in the walls 16-20 h after fertil-
ization has a mannuronic to guluronic acid ratio of 0.33-0.55 and consists of
5 4 ~ alternating units 3 0 ~ of guluronic acid blocks and 16 ~ of mannuronic
acid blocks. Such an alginate would exist as a transparent flexible gel with high
mechanical strength.
Shiraiwa et al. (1975) and Madgwick, Haug & Larsen (1973b) have prepared
alginate lyases from a number of brown seaweeds which cleave the alginate
chains producing 4,5-unsaturated acid at the non-reducing end of the cleaved
chains (Fig. 8).

"FUCANS"

Finally the sulphated polysaccharide of the brown seaweeds has been

variously described as fucoidin, fucoidan, ascophyllan, sargassan and glucuron-
oxylofuean. I believe that it comprises a family of polydisperse heterolnolecules
containing, in addition to fucose, varying proportions of galactose, mannose,
 Polysaccharides of green, red and brown seaweeds 113

xylose a n d glucuronic acid (Percival, 1978c). F o r simplicity I shall call such

p o l y m e r s " f u c a n s " on the u n d e r s t a n d i n g that they contain other sugars as well
as fucose. I n the early studies extensive purification was carried o u t in an a t t e m p t
to isolate a " f u c a n " c o n t a i n i n g only fucose residues. This was never achieved
c o m p l e t e l y a n d even in the so-called purest samples small p r o p o r t i o n s o f
galactose, xylose a n d u r o n i c acid persisted (Percival & Ross, 1950). I
t h i n k t h a t the material finally separated b y these w o r k e r s was one fraction o f
this polydisperse family o f molecules. F r a c t i o n a t i o n experiments we have c a r r i e d
o u t on extracts f r o m a n u m b e r o f different genera o f b r o w n weeds ( J a b b a r - M i a n
& Percival, 1973a) (Table IV) have separated fractions c o n t a i n i n g as m u c h as
20 ~ u r o n i c acid and p r o p o r t i o n a t e l y less fucose a n d sulphate, a n d at the other
e n d o f the f r a c t i o n a t i o n the u r o n i c acid is r e d u c e d to 3 - 4 ~ a n d the fucose a n d
sulphate c o r r e s p o n d i n g l y increase. S u p p o r t for this heterogeneity has been
o b t a i n e d for the fucans f r o m Ascophyllum nodosum a n d Fucus vesiculosus
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( M e d c a l f & Larsen, 1977a).

TABLE IV. Fractionation of "Fucans". On DE-cellulose columns by elution with increasing

concentrations of aqueous potassium chloride

Carbo- Uronic Molar proportions of sugars

[~]D Recovery hydrate Sulphale acid
Species (degrees) (~) (~) (~) (~) Fuc Xyl GIcUA Gal

Himanthalia lorea
KCI fraction :
0.3 M --87 28 50-52 2"5 18"8 2'5 1"9 1"8 +
0.5 M --58 16 45-47 21"0 8"9 10"0 1"0 4"0 -¢-+ -t-
M --132 33 40-42 29"0 4'0 14'0 1-0 2"0 +-i-
Bifurearia bifurcata
KC1 fraction :
03 M --84 42 48-50 4'6 19"5 2'8 1"0 2"4 ÷
0"5 M --54 8 43--45 22"7 11"5 5"5 1"0 2'5 -1--I-
M -- 100 20 40-42 30"0 2"6 13"0 1"0 2'3 + -}-+
Padina pavonia
KCI fraction :
0'3 M 94 31 50-52 2"5 20'4 2'0 l "0 2"0 -1-
0"5 M --51 20 45--47 11"0 9'1 3"0 1"0 1"2 ÷ +
M --112 38 40 42 17"0 4"7 12"5 I'0 30 ++-]-

Quantity of galactose visually determined on paper chromatograms

Structural studies have shown t h a t these polysaccharides c o m p r i s e extremely

c o m p l e x highly b r a n c h e d macromolecules. Evidence o f 1,2-, 1,3- a n d 1,4-1inked
a n d sulphated fucose units have been o b t a i n e d ( J a b b a r - M i a n & Percival, 1973b).
Different groups o f w o r k e r s have a d v a n c e d a variety o f structures for the m a c r o -
molecules including a glucuronic acid b a c k b o n e for the " f u c a n " f r o m Asco-
phyllum n6dosum (Larsen, H a u g & Painter, 1966). L a t e r w o r k ( M e d c a l f &
Larsen, 1977b) by the N o r w e g i a n workers, however, on dilute acid extracted
" f u c a n " complexes f r o m this weed, indicated a variety o f structures for the
different fractions, b u t until m o r e direct evidence is a d v a n c e d their interpreta-
tion o f the m a c r o m o l e c u l a r structures should be accepted with caution.
G l u c u r o n o s y l - m a n n o s e as a r e p e a t i n g unit in a b a c k b o n e for the polysaccharide
f r o m Sargassum linifolium has been advanced ( A b d e l - F a t t a h , Hussein & Salem,
 114 E. PERCIVAL

1974). Here again the results could be interpreted differently. It is safer to

explain most of the results on the basis of the inner or outer part of these highly
branched polysaccharides.

Biosynthesis
Little is known about the synthesis of the "fucans". Bidwell & Ghosh (1963)
postulate that like the galactans of the Rhodophyceae, sulphate is rapidly
exchanged without complete breakdown and resynthesis. This is supported by
Lestang & Quillet (1974) who found that living fronds of Pelvetia canaliculata
immersed in seawater from which sulphate ions had been removed secreted
sulphate into the water and that fronds which had been partially depleted of
sulphate absorbed sulphate from seawater, and they concluded that both
processes were taking place simultaneously. The exchange did not take place
with fronds which had been killed by boiling or prolonged drying, nor did
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absorption take place when respiration was prevented, and they concluded that
the energy of respiration was required to form the sulphate esters. Furthermore,
they isolated a cytidine diphosphosulphate (CDPS), which in association with
manganese they consider to be the intermediary in the enzymic sulphation of the
fucose residues. They postulate that this turnover of sulphate removes toxic
concentrations of sodium ions from the plant tissues.
Further work (Lestang-Bremond & Quillet, 1976) using 3sSO4Z- in seawater
in which sections of frond were immersed, confirmed the sulphate exchange,
and extraction of the "fucan" showed that the 35SO~- was combined in that
constituent of the seaweed. Only about 3.5 ~ of the total sulphate takes part
in the exchange. These authors assumed that only this part of the sulphate,
which they refer to as the sulphate on the outside of the globular "fncan"
molecule, could undergo exchange. In view of the known heterogeneity of
"fucans" and the probability that "fucans" of different composition are associ-
ated with various parts of the seaweed other explanations are obviously possible.
In the blades of Laminaria sp. specialized secretory cells have been shown
(Evans, Simpson & Callow, 1973), by light microscope autoradiography using
35SO4Z-, to be the sole site of synthesis of sulphated polysaccharides. The fucan
is secreted into mucilage canals which pass through the thallus and empty their
contents on to the surface. E. M. autoradiography using 35SO2- have shown the
Golgi complex to be the site of polysaccharide sulphation (Evans & Callow, 1974).
Various enzymes have been shown to be associated with the Golgi-rich
fraction from the vegetative thallus of Fucus serratus, including one which
transfers galactose from UDP-galactose to "fucan" (Coughlan & Evans,
1978). 35SO2- studies have shown that the Golgi bodies are sites of fucan
sulphation in 22 h zygotes of this seaweed (Callow, Coughlan & Evans, 1978).
The sulphate acceptor molecule has been partially characterized as 3sS-APS
and 35S-PAPS.

Functions
Apart from other functions this polysaccharide almost certainly protects the
plant from desiccation. Lestang and Quillet (1974) showed that the "fucan"
extracted from Pelvetia canaliculata had a very strong affinity for magnesium
 Polysaccharides of green, red and brown seaweeds 115

so that when the fronds are in contact with seawater the ester sulphate groups
are largely associated with magnesium ions. They point out that magnesium
ions are highly hydrated and will therefore retain water in the fronds. In
support of this conclusion is the fact that weeds such as Pelvetia canalieulata,
which grow on the higher part of the shores and are therefore frequently
exposed, have a high "fucan" content, whereas Desmarestia and Durvillea
species which grow below low tide mark contain very little or no "fucan"
(Carlberg, Percival & Rhaman, 1978).
"Fucans" are included, like the galactans, in the "Cell wall and intercellular
region polysaccharides" chapter of Algal Physiology and Biochemistry (Mackie
& Preston, 1974), and evidence from electronmicroscope studies indicate that
they are laid down in the vicinity of the cell walls. While a certain proportion of
the "fucan" of Fucus and AseophylIum is extracted by water and dilute acid the
residual weed always contains some "fucan", supporting the fact that at least
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some of the molecules are bound in the cell wall structure. Evidence of this was
indicated by 14C studies (Bidwell, Percival & Smestad, 1972) on Fucus vesieulosus
"fucans". From these it appeared that a xylogalactofucoglucuronan is syn-
thesized first and that this is transformed into a xyloglucuronogalactofucan
(both named in order of the increasing proportions of their constituent sugars)
and a "fucan" containing 90 ~ of fucose units was finally synthesized, each of
these polymers possibly having a different function in the alga.
Quatrano and his colleagues in their elegant studies on the development of
the zygotes of species of Fueus distinguished two types of "fucan" (Quatrano &
Stevens, 1976; Quatrano & Crayton, 1973): F1 which had a negligible sulphate
content and F2 which is fairly highly sulphated. F1 is present in the cell walls
during the first 4 h after fertilization. The authors consider that after 10 h F1
has been enzymically sulphated and forms 20 ~ of the cell walls of the zygote.
Although embryos grown in the absence of sulphate form normal rhizoids
(Crayton, Wilson & Quatrano, 1974) the fucan is not sulphated and the rhizoids
fail to adhere to the substratum.
This must not be regarded as a survey of all the work carried out in this field,
but is a brief description of the topics which the author considers the more
important and interesting. It must be remembered that the earlier studies on
these polysaccharides were carried out before the development of chromato-
graphy, NMR, electronmicroscopy, and many other modern techniques had
been developed. These have made the study of polysaccharides much easier
and less tedious. In the early days gram quantities of pure polysaccharide were
required for any detailed study and considerable credit should be given to the
patience and skill of the early workers in this field. It is only during the last
10-15 years that investigations on milligram and even smaller quantities are
possible. As a result during the forthcoming decade very large advances in the
chemistry and biology of these polysaccharides can be expected.

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