Journal of Applied Microbiology Symposium Supplement 1997, 83, 80S88S

Sugar metabolism by mutans streptococci

S.M. Colby and R.R.B. Russell
Department of Oral Biology, Dental School, University of Newcastle, Newcastle upon Tyne, UK

1.
2.
3.
4.

Introduction, 80S
Oral streptococci, 80S
Carbohydrate metabolism in plaque, 81S
Fructans in dental plaque
4.1 Fructosyltransferase, 82S
4.2 Fructanase, 82S
5. Glucans in dental plaque

6.
7.
8.
9.

5.1 Glucosyltransferase, 83S

5.2 Dextranase, 84S
5.3 Dextranase inhibitor, 85S
Glucan-binding proteins, 85S
Comparisons between species, 86S
Acknowledgements, 86S
References, 86S

1. INTRODUCTION

Dental caries are arguably the most prevalent bacterial disease

in developed countries and are responsible for enormous costs
in terms of pain and discomfort, productivity losses due to
toothache or dental appointments and provision of professional treatment. It is now just over a century since it was
first proposed that caries (dissolution of tooth enamel) is due
to the acids generated by bacterial fermentation of carbohydrates and the link between dietary sugar and dental caries
is well established (Moynihan 1996). While many carbohydrates may be utilized by plaque bacteria to generate acids,
sucrose is recognized as being particularly important in the
caries process because not only can it be fermented, but it
also serves as substrate for extracellular enzymes of plaque
bacteria which synthesize sucrose-derived polymers. These
polymers are of central importance in adhesive interactions
in plaque, where they mediate attachment of bacteria to the
tooth surface and to other bacteria thus stabilizing the plaque
biofilm, serve as energy stores aiding the survival of plaque
bacteria and modulate the permeability of plaque and hence
the level of acid at the enamel surface. Streptococci are of
central importance in the formation and metabolic activity of
plaque, and recent years have seen significant advances in
our understanding of the processes which can lead to the
development of caries.

2. ORAL STREPTOCOCCI

Some 19 distinct species of streptococci have the human oral

cavity as their natural habitat, all of them belonging to the
Correspondence to: Professor R. R. B. Russell, Department of Oral Biology,
Dental School, University of Newcastle, Newcastle upon Tyne NE2 4BW, UK.

viridans group and advances in taxonomy in recent years

(see Hardie and Whiley, pp. 1S11S) have led to an appreciation of their different properties and contribution to dental
plaque formation. Plaque represents a highly complex ecosystem with many different types of bacteria present (Milnes
et al. 1996) but streptococci generally comprise the majority
and so have attracted considerably more research interest
than other plaque species.
On the cleaned tooth surface, Streptococcus sanguis, Strep.
mitis and Strep. oralis predominate among the first bacteria to
colonize and it is believed that these pioneer species help to
establish conditions for the subsequent development of the
plaque biofilm by a combination of processes: growth and
division of attached bacteria to form microcolonies, establishment of other species through coaggregation with the
pioneers (Kolenbrander 1993) and local changes in physiological conditions which encourage proliferation of other
species. Of particular interest are the circumstances which
encourage colonization by Strep. mutans and Strep. sobrinus,
which are the two species clearly implicated in the initiation
of caries. Studies from around the world have shown that
98% of adults carry Strep. mutans, while different studies
have found Strep. sobrinus in 735% of individuals. The
majority of epidemiological studies exploring the association
between streptococci and caries have determined the combined numbers of the two species so their relative importance
is unclear, though in populations with a high caries prevalence
it is obvious that Strep. mutans must be the significant organism since it is mostly found in the absence of Strep. sobrinus.
There have, however, been suggestions that Strep. sobrinus is
more virulent when present (de Soet et al. 1993). Both species,
which belong to the mutans group of streptococci, cause
caries in experimental animals and both have been the subject
of extensive investigation at the molecular level (Russell
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S TR EP T OC OC C AL SU G AR ME T AB OL I SM 81S

1994). Other members of the mutans group of streptococci,

Streptococcus cricetus, Streptococcus rattus and Streptococcus
downei, were originally isolated from animals and are rarely,
if ever, found in humans. These species can, however, provide
useful model organisms for the study of various biological
properties which they share with the species found in
humans.
In healthy adults with a low level of caries experience and
an exemplary diet, i.e. one with a low sucrose consumption
and no snacking between meals, the mutans streptococci may
comprise less than 001% of the plaque bacteria. A switch to
a sugar-rich diet leads to an increase in the mutans species
and mutans streptococci may comprise more than half of the
viable bacteria in plaque at tooth sites where caries are active.
Longitudinal studies have demonstrated that a high level of
mutans streptococci in plaque at a caries-prone tooth site
(particularly in fissures or between teeth) gives a high risk of
subsequently developing caries at that site (Loesche 1986).
We are therefore faced with two questions: what conditions
encourage the mutans streptococci, and what properties make
them so cariogenic once they are established? This article will
not concern itself with the surface components of streptococci
which may be involved in adhesive interactions with other
bacteria or with host macromolecules but will focus on the
physiological processes involving carbohydrates.

3. CARBOHYDRATE METABOLISM IN
PLAQUE

Pioneer species of streptococci are not in general strong acid

producers but, in the presence of fermentable carbohydrate
in the diet, they will lower the pH so that acid-tolerant
bacteria will be favoured. A sugar-rich diet can thus induce
an ecological shift in the plaque microflora and a resultant
increase in aciduric species, especially mutans streptococci
and lactobacilli (Marsh 1994). There is little detailed information available on sugar transport and metabolic pathways
in the pioneer species of streptococci, though it is clear that
even within individual species there is variation in acidproducing capacity. It is thus apparent that some strains of
species which have not shown an overall association with
caries may still be important in the development of caries,
and account for the group of non-mutans streptococci
detected in caries lesions where no mutans streptococci are
detectable (Sansone et al. 1993). Simple classification to the
species level therefore seems to be inadequate in delineating
the properties which may make a bacterium cariogenic and it
is unwise to consider the mutans streptococci to have a monopoly of properties which may contribute to caries. Nevertheless, it seems probable that the virulence properties of all
cariogenic streptococci are comparable, and detailed infor-

mation on certain species will give clues as to the capabilities

of others.
Streptococcus mutans is remarkably versatile in the range of
carbohydrates which it can utilize and this may be one of the
features which enables it to outgrow other species when the
diet is rich in carbohydrate, regardless of the particular sugars
present. Versatility of Strep. mutans is also manifested with
regard to sugar uptake and aciduricity. Although the oral
cavity might be considered to be an ideal place for bacteria
to grow, being warm, moist and well supplied with nutrients
(host salivary glycoproteins supplemented with three or more
meals a day), local conditions in dental plaque are subject
to considerable fluctuation. The main continuous source of
sugars comes from the breakdown of saliva but during meals
it has been estimated that the available sugar may rise 100 000fold from 1 mmol l1 to 100 mmol l1 while the pH may vary
from greater than 67 (the average pH of saliva) to less than
pH 40. In order to flourish in such challenging conditions
Strep. mutans has a multiplicity of transport mechanisms. For
example, sucrose is taken up by three distinct systems with
Km values of 71 105 mol l1, 25 104 mol l1 and
33 103 mol l1 so that there is efficient uptake over a
broad range of external sucrose concentrations (Slee and
Tanzer 1982). The uptake systems have been identified as
a sucrose-specific phosphotransferase (PTS), the trehalosePTS and a third transport system which may correspond to
the multiple sugar metabolism system (Tao et al. 1993). Once
inside the cell, sugars are fermented by the glycolytic pathway
and Strep. mutans can maintain glycolysis at pH values as low
as 38. Streptococcus sobrinus is even more aciduric, though it
is capable of utilizing a narrower range of sugars than Strep.
mutans. The molecular basis of aciduricity of the mutans
streptococci is not yet understood but is the subject of investigation in a number of laboratories.
Clearly, any of the sugars which can be fermented may
contribute to acid production but of particular interest is the
metabolism of the sugar which has been termed the archcriminal in dental caries, sucrose. The levels of mutans streptococci in the mouth respond rapidly to changes in the
amount of sucrose in the diet (Wennerholm et al. 1995) and
a number of properties may explain this. Direct uptake of
sucrose has already been alluded to above, and it has been
estimated that 90% of the sucrose encountered by Strep.
mutans is taken up and enters glycolysis, leading to acid
generation. However, since this pathway is common to all
sugars, the intracellular metabolism of sucrose may be considered to offer no insights into its unique propensity to
contribute to caries. On the other hand, the metabolism of
sucrose by extracellular enzymes does offer some distinctive
features and its different fates are summarized in Fig. 1.
Extracellular enzymes synthesizing fructans or glucans,
enzymes degrading these polymers and glucan-binding proteins have all been identified.

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82S S .M . C O LB Y A N D R .R . B. RU S SE LL

(a) Strep. mutans

gtf B

modified
glucans

glucans

gtf C

sucrose

dexA

gtf D

+
ftf

fructose

isomaltosaccharides

fructan +
msm
glucose

fruA

dexB

fructose

glucose

(b) Strep. sobrinus

gtf I

modified
glucans
dei

glucans

gtf S
dexA

gtf T

sucrose

cleaves sucrose to release free glucose (which is available for

uptake into the cell) while linking the fructose parts of the
molecule together to yield an inulin-like fructan. Until the
introduction of gene cloning techniques, it was not clear how
many FTF Strep. mutans were produced because multiple
bands of FTF activity ranging in size from 95 000 to
79 000 Da could be resolved by SDS-PAGE (Russell and
Gilpin 1987). A single ftf gene has now been cloned and
nucleotide sequencing has shown it to have a deduced molecular weight of 87 600 (Shiroza and Kuramitsu 1988). The
reason for the appearance of multiple electrophoretic forms
is not clear, but post-translational modification by proteolysis
is a common feature of the extracellular enzymes of Strep.
mutans (Russell et al. 1986). Streptococcus sobrinus does not
have a fructosyltransferase.

4.2 Fructanase
+

gtf U

+
isomaltosaccharides
fructose

Fig. 1 The metabolism of sucrose by extracellular enzymes in

(a) Streptococcus mutans and (b) Strep. sobrinus. Gene symbols:

gtf, glucosyltransferase; dexA, dextranase; ftf,
fructosyltransferase; fruA, fructanase; msm, multiple sugar
metabolism operon; dexB, dextran glucosidase; dei, dextranase
inhibitor. Both species also have mechanisms for the direct uptake
of sucrose

4. FRUCTANS I N D ENTAL PLAQUE

Fructans are linear polymers consisting solely of fructose

units, synthesized from sucrose. The fructose units may be
joined by b(26)-linkages, in which case they are referred to
as levan, or b(21)-linkages as are found in inulin from
dahlia tubers. Despite evidence for a high rate of synthesis of
fructan, only low levels are found in plaque and this is
believed to be due to a high rate of turnover, with fructan
being rapidly degraded. Fructan thus serves as a short-term
extracellular energy storage molecule. There is conflicting
evidence as to whether the ability to make fructan contributes
to the virulence of Strep. mutans (Kuramitsu 1993).
4.1 Fructosyltransferase

In Strep. mutans, the most active extracellular enzyme acting

on sucrose is fructosyltransferase (FTF), an enzyme which

It is likely that a variety of different plaque bacteria can

exploit fructan as a substrate and Strep. mutans itself has a
fructanase. This enzyme is an exofructosidase which releases
single fructose units which can be taken up and metabolized.
Fructanase is found in the supernatant fluid of liquid cultures
so an unexpected finding from the sequencing of the fruA
gene was the presence of an LPXTG motif close to the Cterminus (Burne and Penders 1992). Such a motif, together
with a C-terminal hydrophobic tail, is characteristic of proteins which are covalently anchored to peptidoglycan. Two
wall-associated proteins of Strep. mutans (Ferretti et al. 1989;
Okahashi et al. 1989) and the dextranase described below also
have these wall-anchor motifs and a region rich in proline or
serine threonine which is thought to correspond to a wallspanning segment. Although LPXTG proteins may be covalently linked to peptidoglycan (Schneewind et al. 1995) all the
Strep. mutans proteins with this motif are released, the extent
of release depending upon growth conditions (Lee 1992;
Burne and Penders 1994). The surface protein WapA has a
site of cleavage outside the wall-spanning layer (Ferretti et
al. 1989) and fructanase also is subject to proteolysis (Burne
and Penders 1994) but the sites of cleavage have not been
defined. There may be more than one release mechanism
and the biological significance of the phenomenon remains
unclear. It has been suggested that the release of surface
components by a surface protein releasing enzyme (SPRE)
can modulate the balance between immobilized cells and
planktonic ones which would be free to migrate and colonize
other sites in the mouth (Lee et al. 1996). An alternative
viewpoint is to envisage varying conditions in plaque when
there may be different advantages to Strep. mutans of having
enzymes such as fructanase surface-associated (benefiting the
single cell) or a roving fructanase which would benefit the
entire local microbial community.

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5. GLUCANS IN DENTAL PLAQUE

In contrast to the rapid turnover of fructan, glucans are a

constant feature of plaque (Hotz et al. 1972) and consist of
glucose units joined by a(16)- and/or a(13)-linkages. The
relative proportions of these two linkages and the degree of
branching determine the ultimate properties of the glucan. A
glucan which is essentially a linear a(16)-linked chain is
referred to as a dextran and is soluble (the trivial name
dextransucrase is often applied to the enzyme which produces
dextran); in contrast mutan is a water-insoluble glucan with
a high proportion of a(13)-linkages. In practice, the glucan
in plaque will be a highly complex macromolecular network
resulting from the action of synthetic and degradative
enzymes described below. However, experiments have shown
that both soluble and insoluble glucans are important in cell
cell and cellsurface adhesive interactions in dental plaque,
with dextran-mediating bacterial aggregation while a(13)rich, branched insoluble glucans have been shown to be the
major contributor to adherence to hard surfaces (Ebisu et al.
1974).
Besides their adhesive properties, a range of other functions for glucans has been suggested, including acting as an
extracellular energy store (see below) and protecting streptococci from attack by host defences, bacteriophage or bacteriocins. With regard to the caries process, it has been
proposed that their major significance may be in establishing
an intercellular matrix which determines the density of bacterial cells in plaque: in the presence of sucrose large amounts
of glucan are produced and effectively reduce the number of
Strep. mutans in a unit volume. However, the looser plaque
structure results in more efficient diffusion of nutrients
through the matrix and this leads to an overall greater metabolic activity and hence higher rate of acid production

Table 1 Glucosyltransferases of

Streptococcus sobrinus

(Dibdin and Shellis 1988; van Houte et al. 1989). Exposure

to sucrose thus has a number of effects, aiding colonization
by adhesion to the tooth and increasing the total plaque bulk
while at the same time enhancing the overall rate of acid
production

5.1 Glucosyltransferase

Because of the importance of glucans in plaque, the glucosyltransferases (GTF) which make glucans have attracted
considerable interest. Streptococcus mutans produces three distinct GTF while Strep. sobrinus produces four. Each GTF is
encoded by a separate gene and each enzyme has distinctive
properties, varying in its requirement for a primer molecule
to start the polymerization reaction, the proportion of a(1
6)- and a(13)-linkages and degree of branching it introduces
into the glucan, and the total length of the glucan chain
produced. The glucans made by the individual GTF of Strep.
mutans have not been well characterized, but the GTF of
Strep. sobrinus all have individual characteristics (Table 1).
Within dental plaque, the overall properties of the glucan
present will depend on the relative activity of the different
GTF present and also on their interactions, since one GTF
may modify the product of another.
The gene sequences of 12 streptococcal GTF are now
available, as well as two from Leuconostoc mesenteroides. Multiple alignment of the deduced amino acid sequences has
revealed common features of GTF. All are of high molecular
weight (155174 kDa) and composed of distinct domains
(Fig. 2). The challenge facing those working in the field of
GTF is to identify those features of the protein structure
which determine function. We need to understand what it is
that determines why a particular GTF makes a particular

Glucosyltransferase
Glucan

Enzyme
Water
Molecular
Gene names
Primer
Structure
solubility
weight

gtf I
I* P3 Dependent
Branched,
Insoluble
High
mostly, a(13)-linkages
(insoluble)
gtf U S1 P4
Dependent
Branched,
Soluble
100 000
a(16)- and
a(13)-linkages
gtf S
S3 P2
Independent
Linear,
Soluble
5300
a(16)-linkages
gtf T
S4 P1
Independent
Branched,
Soluble
400 000
a(16)- and
a(13)-linkages

Nomenclature used by Cheetham et al. (1991)* and Hanada et al. (1993). Data on
primer dependence and glucan properties are also taken from these authors.

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84S S .M . C O LB Y A N D R .R . B. RU S SE LL

type of glycosyl linkage, what determines the length of glucan

chain which it produces and what influences the kinetics of
the enzyme reaction, such as the need for a primer glucan.
Detailed knowledge of the individual GTF properties will
help in elucidating the way in which they interact with each
other.
Once the deduced amino acid sequence of GTF became
available, a search of the databanks indicated a degree of
relatedness to a-amylase (Ferretti et al. 1987) and suggested
that comparison with other enzymes acting on carbohydrate
substrates would be informative. GTF can be regarded as
glycosyl hydrolases, which have the ability to hydrolyse
sucrose and also to transfer the released glucose to a growing
acceptor chain. Information on other glycosidic enzymes
should thus aid our understanding of GTF. For example, it
is known that the catalytic mechanism of glycosidases involves
glutamic acid or aspartic acid residues, providing a nucleophilic carboxylate and a general acid catalyst (Sinnot 1990).
While multiple alignment of related sequences can often help
identify conserved residues essential for catalysis, alignment
of GTF sequences has failed to locate the critical residues
(there are 23 points where glutamate is conserved in all GTF!)
so progress depends on advances in structural determination
or comparison with heterologous sequences. Mooser et al.
(1991) were able to trap a glucosyl-enzyme intermediate formed during the reaction of GTF with sucrose and isolated
the relevant active site peptide from GTF, demonstrating
that it was homologous to a common glycosidase motif, with
an aspartic acid implicated as the core residue. Kato et al.
(1992) have confirmed the importance of this residue by
site-directed mutagenesis. From our knowledge of enzyme
mechanisms, we can anticipate that a number of amino acid
residues well separated on the primary sequence but juxtaposed when the protein is folded into its tertiary structure
will play critical roles in GTF function. The large size of
GTF makes them extremely challenging molecules for crystallization and X-ray crystallography and no 3-D structural
model is yet available. However, there has been a recent
exciting advance in determining the structure of GTF by
MacGregor et al. (1996), who applied a combination of
sequence alignment, hydrophobic cluster analysis and structure prediction strategies to GTF.
Glycosyl hydrolases have been grouped and classified into
a series of families on the basis of amino acid sequence similarities and, as structures become available, larger groupings
of structurally related families are being formed (Davies and
Henrissat 1995). One major grouping consists of enzymes
with a (b/a)8 barrel structure also referred to as a TIM
barrel in which eight b strands (E1E8) alternate with eight
a helices (H1H8). Included in this group are a-amylase,
cycloglucanotransferase, isoamylase and the DexB dextran
glucosidase from Strep. mutans. The sequences of all these
proteins can be aligned on the basis of their b sheets and a

A B

Fig. 2 Representation of the domain structure common to all

glucosyltransferases (GTFs). A, Signal peptide (42% homology

between GTFs); B, variable region of unknown function (ca 200
residues) which is unique to each GTF; C, conserved region (43%
homology between GTFs) of ca 800 residues. This is thought to
be the catalytic domain. The solid bar indicates the region
homologous to the (b/a)8 barrel; D, repeat region essential for
glucan binding. This domain consists of a series of tandem
repeats (ca 30 amino acids each) which are also found in proteins
of other organisms and may represent a common type of sugar
binding domain

helices and the central domain of GTF sequences can also be

aligned in this way (Fig. 2). The striking and distinctive
feature of the GTF alignment is that the first helix detected
in the GTF sequence aligns with a helix H3 of the other
proteins, while the b and a segments normally found at the
start of proteins such as amylase (E1 H1 E2 H2 E3) come in
the latter part of the GTF sequence. In other words, in the
GTF there is a totally novel construction in which the usual
arrangement of alternating sheets and helices has undergone
cyclic permutation (MacGregor et al. 1996). Nevertheless, all
the elements required to form the predicted standard (b/a)8
barrel are present in GTF and this model has the potential
of a major advance in understanding GTF structure/function
relationships. There is now a considerable body of knowledge
on the amino acid residues involved in the active site of
amylase and some of the other glycosidases, so superimposition of the amylase and GTF sequences allows prediction of which residues are important in GTF. Although
amylases show very great sequence diversity, four well conserved regions have been identified and the corresponding
regions of GTF can be recognized. Site-directed mutagenesis
of specific amino acids can now proceed on a rational basis
and yield new insights into GTF mechanisms.
5.2 Dextranase

The nature and amount of glucan in plaque is influenced not

only by the activity of the GTFs but also by that of extracellular dextranase. A wide range of bacterial species associated with dental plaque have been shown to produce
dextranase (Johnson 1990) and those of Strep. mutans and
Strep. sobrinus have been studied in some detail (Wanda and
Curtiss 1994; Igarashi et al. 1995).
Dextranase (Dex) is able to break down glucans to isomaltosaccharides 34 glucose units long by cleaving a(16)linkages within the dextran chain (Pulkownik and Walker
1977; Walker et al. 1981). This activity may modify glucans

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S TR EP T OC OC C AL SU G AR ME T AB OL I SM 85S

by altering the ratio of a(16)- to a(13)-linked chains, hence

influencing solubility and adhesive properties. Dex activity
therefore influences sucrose-dependent adherence by reducing the number of a(16)-linkages in the glucan and this has
been demonstrated in Strep. mutans where Dex mutants
accumulate onto a smooth surface to a markedly greater extent
than the wild type (Colby et al. 1995). A further consequence
of Dex activity may be to provide primer or branch points
for GTF thus contributing to the complexity of the glucan
structure (Germaine et al. 1977).
Apart from its effect on the adhesiveness of glucans, the
dextranase of Strep. mutans breaks down glucans to isomaltosaccharides which may then be transported into the cell
via the multiple sugar metabolism (msm) operon (Russell et
al. 1992; Tao et al. 1993) where they are further degraded to
glucose by a dextran glucosidase, DexB (Colby et al. 1995).
Streptococcus sobrinus does not have such an uptake mechanism and is unable to utilize dextrans or isomaltosaccharides
as sole carbon sources (Ellis and Miller 1977).
There is significant homology between the deduced amino
acid sequences of Strep. mutans and Strep. sobrinus dextranases. Both have N-terminal signal peptide sequences as would
be expected of extracellular enzymes and, near the C-terminal
end, the LPXTG motif followed by a hydrophobic region
and a charged tail similar to the proposed wall-anchor region
in fructanase (Fig. 3). Dextranase differs from other LPXTG
proteins in not having a clearly defined region rich in the
hydrophilic amino acids serine, threonine or proline but it is
remarkable in having a large portion of the molecule apparently not needed for dextranase activity. The size of the
dextranases differs and it seems reasonable to assume that
the common region is responsible for dextranase activity.
Multiple active forms of Dex have been reported for Strep.
sobrinus, ranging in size from 130 000 to 81 000 (Barrett et al.
1987) and a similar phenomenon is seen with Strep. mutans
(Russell and Ferretti 1990; Igarashi et al. 1992; Colby et al.
1995) and also Strep. salivarius (Lawman and Bleiweis 1991).
The points of cleavage have not been determined but it seems
likely that the multiple forms result from sequential removal
of C-terminal stretches of amino acids. However, the role of
the non-enzymatic C-terminal regions of these large molecules remains an enigma.

Strep. mutans Dex

Strep. sobrinus Dex

A B

Fig. 3 Representation of the structure of the extracellular

dextranase enzymes of Streptococcus mutans and Strep. sobrinus. Both

proteins possess a signal peptide (A), a conserved domain with
57% homology (C), and two variable regions (B and D)

5.3 Dextranase inhibitor

Despite the similarity in amino acid sequence, there appear

to be differences in the regulation of dextranase in the oral
streptococci. A heat-stable protein which inhibits dextranase
activity, known as Dei, has been detected in Strep. sobrinus
and Strep. downei but not in Strep. mutans (Hamelik and
McCabe 1982; Sun et al. 1994). The activity of this protein
is specific since Dei from Strep. sobrinus will only inhibit
dextranase from Strep. sobrinus and the closely related Strep.
downei and Strep. cricetus but not from Strep. mutans (Pearce
et al. 1991; Sun et al. 1994). Under conditions of carbohydrate
limitation of Strep. sobrinus, Dei levels are high and so little
active dextranase can be detected. When growth rates
increase, however, as would occur in plaque at meal times,
the relative proportions and binding of Dex and Dei alter
so that under these conditions free Dex becomes available
(Wellington et al. 1994). This contrasts with Strep. mutans
where available Dex is released from early exponential phase
(Walker et al. 1981).

6. GLUCAN-BINDING PROTEINS

The glucan-binding domain of GTF may have dual functions: at least one or two of the repeat units are needed for
glucan synthesis (Ferretti et al. 1987) but the binding capacity
will also lead to glucan-mediated binding of bacterial cells
through surface-associated GTF. Free GTF is also found in
saliva and on the tooth surface where it remains active so that
glucans made on the tooth form another site of attachment
for bacteria (Schilling and Bowen 1992). Besides GTF, a
number of other glucan-binding proteins (GBP) without
known enzyme activity can be found. Streptococcus mutans
produces two GBPs, one of which consists of a series of
tandem repeat units homologous to those found in the carboxyterminal one-third of GTF (Banas et al. 1990). The
occurrence of this independent GBP is consistent with the
idea that GTF have a modular structure and this is supported
by the fact that the Dei dextranase inhibitor also contains
homologous repeat units (Sun et al. 1995). Furthermore,
homologous sequences are found in proteins from other
organisms which also have a modular structure: autolysin
of Strep. pneumoniae, fibronectin-binding protein of Strep.
pyogenes and toxin A of Clostridium difficile (Wren 1991; Talay
et al. 1994; Yother and White 1994). In all these cases the
repeat-containing module is thought to be involved in binding
to a macromolecular carbohydrate substrate, and subtle differences in protein sequence must determine the specificity
of binding.
A second GBP which induces a strong immune response
in children has been described in Strep. mutans (Smith et al.
1994) and Strep. sobrinus also produces a number of GBP (Ma
et al. 1996; Wu-Yuan and Gill 1996). Ma et al. (1996) have

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86S S .M . C O LB Y A N D R .R . B. RU S SE LL

produced evidence that only certain of these, which they call

glucan-binding lectins (GBL), can mediate aggregation by
glucan.

7. COMPARISONS BETWEEN S PECIES

Streptococcus mutans and Strep. sobrinus occupy a similar ecological niche and both share the capacity to produce extracellular polymers from sucrose and to adhere to hard surfaces
by means of these polymers. Nevertheless, as Fig. 1 illustrates, the details of these processes are distinct in each species. Streptococcus mutans has three GTF and an FTF as
compared to four GTF (and no FTF) in Strep. sobrinus. While
both species produce dextranase, the controlling inhibitor Dei
is found only in Strep. sobrinus while Strep. mutans can not
only degrade dextran but also utilize the resultant oligosaccharides for intracellular metabolism. The number and
size of GBP also differs between the two species but the way in
which these are involved in adhesion is still unclear. Further
investigations of the function of all these different components should throw light on their relative contributions to
the complex process of growth and adherence in the oral
cavity. In particular, now that many of the genes have been
cloned, targeted gene inactivation allows comparison of
mutants with wild type in in vitro models of adhesion (Colby
et al. 1995) and in vivo, where the capacity to cause caries
in experimental animals can be explored (Kuramitsu 1993).
Improved knowledge of the structure of critical molecules
also sets the scene for rational design of specific inhibitors,
or identification of targets for vaccine development (Taubman
et al. 1995).
This review has presented a number of instances which
illustrate instructive parallels between research on oral streptococci and subjects in quite different areas. Examples are
the structural similarities between GTF of streptococci and
Leuconostoc mesenteroides (the commercial source of dextran),
between GTF and amylase, the finding of homologous carbohydrate binding domains in bacteria from different habitats,
and differing mechanisms of cell wall anchorage in proteins
which initially appear to have common features. Studies of
the oral streptococci thus have much to contribute to, as well
as to learn from, a wide range of microbiological systems.

8. ACKNOWLEDGEMENTS

Research in the authors laboratory has been supported by

the Medical Research Council (Grant Nos. 8504830 and
G9007878CB) and Public Health Service grant DE08191
from the US National Institutes of Health, in collaboration
with the University of Oklahoma.

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