Location via proxy:   [ UP ]  
[Report a bug]   [Manage cookies]                

9661

Download as pdf or txt
Download as pdf or txt
You are on page 1of 12

See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/23307943

Osmoadaptation mechanisms in prokaryotes: Distribution of compatible


solutes

Article in International Microbiology · October 2008


DOI: 10.2436/20.1501.01.55 · Source: PubMed

CITATIONS READS

155 731

2 authors:

Nuno Empadinhas Milton S da Costa


University of Coimbra University of Coimbra
111 PUBLICATIONS 1,332 CITATIONS 294 PUBLICATIONS 8,080 CITATIONS

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

The toxinogenic gut microbiome in sporadic Parkinson’s Disease: a quest for “antiPDbiotics” View project

Microcare - Microbiome of diabetic foot ulcers: early diagnosis, prognosis and therapy View project

All content following this page was uploaded by Milton S da Costa on 22 February 2016.

The user has requested enhancement of the downloaded file.


REVIEW ARTICLE
INTERNATIONAL MICROBIOLOGY (2008) 11:151-161
DOI: 10.2436/20.1501.01.55 ISSN: 1139-6709 www.im.microbios.org

Osmoadaptation mechanisms in prokaryotes:


distribution of compatible solutes
Nuno Empadinhas, Milton S. da Costa*
Department of Biochemistry and Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal
Received 26 May 2008 · Accepted 15 July 2008

Summary. Microorganisms respond to osmotic stress mostly by accumulating compatible solutes, either by uptake from
the medium or by de novo synthesis. These osmotically active molecules preserve the positive turgor pressure required for
cell division. The diversity of compatible solutes is large but falls into a few major chemical categories; they are usually small
organic molecules such as amino acids or their derivatives, and carbohydrates or their derivatives. Some are widely distrib-
uted in nature while others seem to be exclusively present in specific groups of organisms. This review discusses the diver-
sity and distribution of known classes of compatible solutes found in prokaryotes as well as the increasing knowledge of the
genes and pathways involved in their synthesis. The alternative roles of some archetypal compatible solutes not subject to
osmoregulatory constraints are also discussed. [Int Microbiol 2008; 11(3):151-161]

Key words: osmoadaptation · osmoregulation · compatible solutes · trehalose synthesis · biosynthetic pathways · prokary-
otic evolution

Introduction ganisms is inversely proportional to the concentration of dis-


solved solutes [10]. Strategies evolved by microorganisms to
According to a myth, when the Romans destroyed Carthage adjust to high external solute concentrations involve the
in 146 BC, they also salted the ground of the city. Although accumulation of intracellular solutes to counteract the osmotic
this tale seems to have arisen in the nineteenth century, salt stress that might otherwise lead to loss of cellular turgor
has long been associated with death and the infertility of soil pressure, dehydration, and death [4]. Two different strategies
and water—the name “Dead Sea” being one example of the for osmoadaptation have been identified: a reliance on the
ancient idea that large amounts of salt are lethal. This review influx of ions from the surrounding environment (the “salt-in”
deals with organisms that live in water “with a pinch of salt.” strategy), and the accumulation of low-molecular-weight
In addition to providing an environment for biochemical organic compatible solutes to balance the external osmotic
reactions, water may also actively participate as a key pressure.
reagent. However, the amount of water available to microor- The salt-in strategy of prokaryotes seems to be restricted
to: (i) the extremely halophilic Archaea of the family
Halobacteriaceae, which includes the extreme halophiles of
*Corresponding author: M.S. da Costa genera such as Halobacterium, Haloarcula, Haloquadratum,
Departamento de Bioquímica Halorhabdus, Natronobacterium, and Natronococcus; (ii)
Universidade de Coimbra the halophilic Bacteria of the order Haloanaerobiales, and
3001-401 Coimbra, Portugal
Tel. +351-239824024. Fax +351-239855789
(iii) the bacterium Salinibacter ruber [37]. Remarkably, K+ is
E-mail: milton@ci.uc.pt the major cation accumulated, even though, unlike Na+, it is
relatively scarce. The saline cytoplasm of these microorgan-
This article is based on a lecture given by Milton S. da Costa at the Institute
isms requires that most of their enzymes are enriched in
for Catalan Studies, Barcelona, on February 7, 2008, as one of the
Distinguished Lectures of the academic year 2007–2008. MSdC is the pres- acidic amino acids and, at the same time, they are strictly
ident of the Federation of European Microbiological Societies (FEMS). dependent on K+ and/or Na+ for activity.
EMPADINHAS, DA COSTA
152 INT. MICROBIOL. Vol. 11, 2008

The other strategy involves the accumulation of low- neutralization of K+ is accomplished by the accumulation of
molecular-weight organic compatible solutes. Most microor- a few organic anions—mostly amino acids and derivatives—
ganisms have not evolved extensive genetic alterations for such as a-glutamate (Fig. 1) or its isomers, which can be syn-
adaptation to highly saline environments and their cytoplasm thesized or captured from the medium [4]. In general, the
does not tolerate salt. However, this strategy provides a highly accumulation of α-glutamate reaches a physiological plateau
versatile means for adaptation to osmotically changing envi- prior to the activation of osmoadaptive phenomena: the rarer
ronments, as reflected by the large variety of microorganisms β-glutamate has been identified in marine bacteria and some
relying exclusively on organic compatible solutes for methanogenic Archaea [30,42]. The combination of K+ and
osmoadaptation [4]. The accumulation of organic compatible glutamate accumulation seems to be an adequate response to
solutes, within intrinsic limits, is a widespread response in low levels of salt stress [4]. At higher salinities, additional
the microbial world. compatible solutes are required to balance the osmotic pres-
In the natural environment, compatible solutes can be sure and intracellular milieu. It must be noted that Petrotoga
released upon death of organisms or during efflux processes, miotherma, a thermophilic bacterium of the order Thermo-
rendering these compounds accessible to others that can togales, accumulates α-glutamate at the highest salinity tol-
scavenge them for osmoadaptation or as carbon source, pro- erated but accumulates only small amounts of this amino acid
vided they have the appropriate mechanisms for their uptake at low salinities [17]. On the other hand, the closely related
and catabolism [39]. In some cases, the uptake systems are species P. mobilis accumulates not only α-glutamate but also
crucial for microorganisms that do not have the machinery to the β-isomer near the maximum salinities tolerated (our
synthesize appropriate compatible solutes. In addition, a sud- unpublished results). Both isomers of glutamate have been
den dilution of the environment by rain or flooding triggers detected also in the hyperthermophilic bacterium Aquifex
the release of compatible solutes [39]. It is, therefore, com- pyrophilus under salt stress, along with other negatively
mon to find high affinity transport systems in microorgan- charged phosphodiester (see below) compatible solutes [25].
isms that utilize these componds, as they allow them to ade- Alanine, glutamine, and proline (Fig. 1) are other major
quately and rapidly manage the intracellular levels of com- compatible solutes present in several organisms. Some gram-
patible solutes [21]. positive bacteria accumulate low levels of alanine and gluta-
This review will summarize the diversity and distribution mine but they can accumulate proline at very high concentra-
of known classes of compatible solutes in prokaryotes. It will tions. In some members of the genus Streptomyces, the three
also refer to some archetypal compatible solutes that have amino acids accumulate under salt stress [23]. The relatively
escaped osmoregulatory constraints and are used for alterna- low solubility of α-glutamine (compared to the other solutes)
tive purposes, either functional or structural. The molecules accounts for the concentrations almost reaching saturation
studied thus far are discussed with respect to their chemical detected in certain members of the genus Corynebacterium
structure: amino acids; trehalose and sucrose; phosphodi- [10]. The β-isomer of glutamine, which has only been iden-
esters; cyclic-bisphosphoglycerate; mannosylglycerate and tified in halophilic methanogens, can reach very high levels
mannosylglyceramide; glucosylcerate, glucosyl-glucosylcer- in some species; due to its higher solubility, it acts as a very
ate and mannosyl-glucosylglycerate; and polyols. Table 1 efficient compatible solute in osmoadaptation [41]. FIG 1
lists the compounds that have been described to act as com- None of the above solutes surpass proline as a compatible
patible solutes, and the microorganisms or groups of solute in moderate salt stress. In fact, this amino acid can accu-
microorganism in which they have been found. mulate to concentrations in the molar range and may represent
up to 20% of the dry weight of organisms that accumulate it [4].
Within the prokaryotes, proline was initially identified in sever-
Amino acids: widely distributed com- al halophilic members of the genus Bacillus as well as in the
patible solutes in prokaryotes non-halophilic Bacillus subtilis. Only later was it shown that
halophilic/halotolerant Bacillus strains predominantly accumu-
Intracellular K+ contributes to both the osmotic balance late ectoine (Fig. 1), alone or in combination with proline [10].
across the membrane and the stabilization of the cellular tur- Bacillus subtilis seems to represent a minority of gram-positive
gor pressure. In many Bacteria, it increases rapidly with the bacteria that accumulate proline and are unable to synthesize
salinity of the growth medium. However, the incoming other compatible solutes. It can be argued that proline is, in fact,
charge of K+ is not compensated by the accumulation of Cl− an effective compatible solute of halotolerant or moderately
as it is, for example, in the extreme halophilic organisms of halophilic organisms, but not in those living at extreme salt con-
the Halobacteriaceae and Haloanaerobiales [34]. Instead, the centrations. The moderately halophilic bacterium Halobacillus
COMPATIBLE SOLUTES IN PROKARYOTES
INT. MICROBIOL. Vol.11, 2008 153

Table 1. Distribution of compatible solutes in prokaryotes

Compounds Occurrence Ref.


Aminoacids

Nε-Acetyl-β-lysine Unique to methanogenic archaea [38,41]


N-δ-Acetyl-ornithine Bacillus spp., Planocococcus citreus, Sporosarcina halophila [58]
Alanine Streptomyces [23]
Ectoine and hydroxyectoine Halophilic/halotolerant Bacillus strains, Ectothiorhodospira halochloris, aerobic [9,10,51]
heterotrophic bacteria, most halophilic proteobacteria, Micrococcus spp.,
Bacillus spp., Marinococcus spp., Halobacillus halophilus

α-Glutamate Some methanogenic archaea, marine bacteria, Petrotoga miotherma, P. mobilis, [17,25,31,42,50]
Aquifex pyrophilus, Halobacillus halophilus

α-Glutamine Streptomyces, Corynebacterium sp., Halobacillus halophilus [10,23,50]


β-Glutamine Halophilic methanogens [41]
Glycine betaine Bacteria, Archaea (universal compatible solute)a [4]
Proline Streptomyces, halophilic/halotolerant Bacillus strains, halophilic Bacillus strains, [10,23]
B. subtilis (non-halophilic)

Sugars
Sucrose Anabaena, Synechocistis, Nitrosomonas europaea [3,29,40]
Trehalose Corynebacterium glutamicum, Mycobacterium tuberculosis, Thermus [5,7,57]
termophilus, Rubrobacter xylanophilus

Phosphodiesters
Di-myo-inositol phosphate Pyrococcus woesei, Aeropyrum, Aquifex, Archaeoglobus, Pyrodictium, Pyrolobus, [7,49,53]
Stetteria, Thermococcus, Thermotoga, Rubrobacter xylanophilus, Persephonella marina

Di-mannosyl-di-myo-inositol phosphate Thermotoga spp. [30]


Glyceryl-myo-inosityl phosphate Aquifex pyrophilus, Archaeoglobus fulgidus [25,30]
Glyceric acid derivatives
Cyclic-2,3-bisphosphoglycerate Methanothermus fervidus, Methanobacterium thermoautotrophicum, [12,26,55]
Methanopyrus kandleri

Mannosylglycerate Thermus thermophilus, Rhodothermus marinus, Rubrobacter xylanophilus, [7,36,48]


Pyrococcus, Palaeococcus, Thermococcus, Archaeoglobus, Aeropyrum, Stetteria

Mannosylglyceramide Rhodothermus marinus [56]


Glucosylglycerate Agmenellum quadruplicatium, Erwinia chrysanthemi, Persephonella marina [13,49]
Glucosyl-(1,6)-glucosylglycerate Persephonella marina [49]
Mannosyl-(1,2)-glucosylglycerate Petrotoga miotherma [17]
Polyols
Sorbitol Zymomonas mobilis [27]
Mannitol Pseudomonas putida [22]
Glucosylglycerol Pseudomonas mendocina, Stenotrophomonas rhizophila, Synechocystis [16,32,33,43]
a
Glycine betaine spans the three domains of life, even though in many prokaryotes it is not synthesized de novo (see text).

halophilus synthesizes glutamate and glutamine at salinities of Glycine betaine (Fig. 1) seems to be the truly universal
about 1 M NaCl [50]. It must be noted that this organism compatible solute, as it spans the three domains of life, from
switches the osmolyte pool mostly to proline when the salinity members of Bacteria and Archaea, with diverse ecophysio-
of the medium increases to 2–3 M NaCl. logical characteristics, to halotolerant plants and algae.
EMPADINHAS, DA COSTA
154 INT. MICROBIOL. Vol. 11, 2008

Int. Microbiol.

Fig. 1. Structure of some naturally occurring


amino acid compatible solutes and amino
acid derivatives.

Although the role of glycine betaine as a compatible solute is coccus citreus and Sporosarcina halophila [58]. The related
unquestionable, the ability of organisms to synthesize this Nε-acetyl-β-lysine (Fig. 1), unique to methanogenic archaea,
organic solute is not as broadly distributed as initially sus- is produced only under salt stress. The genes involved in the
pected. In fact, many of the regular components of culture synthesis of Nε-acetyl-β-lysine from α-lysine, ablA and ablB,
media, such as yeast extract, contain glycine betain. This ver- encoding lysine-2,3-aminomutase and β-lysine acetyltrans-
satile solute is efficiently taken up by many organisms that ferase, respectively, have been identified on the genomes of
use it to cope with salt stress [4]. Additionally, the term several methanogenic archaea and are co-expressed in a salt-
“biosynthesis” of glycine betaine was frequently used to dependent manner [38].
describe the oxidation of its precursor choline, which is now The compatible solute ectoine and its derivative hydrox-
accepted to be a conversion reaction instead of de novo syn- yectoine (Fig. 1), which were identified in the phototrophic
thesis. The synthesis of glycine betaine is, in fact, a rare phe- sulfur bacterium Ectothiorhodospira halochloris, can be
nomenon in heterotrophic bacteria but common in pho- classified as cyclic forms of N-acetylated amino acids [9]
totrophic bacteria and in methanogenic archaea, which (see their distribution, shown in Table 1). In Halobacillus
exhibit moderate to high salt tolerance. halophilus, ectoine is mainly produced at very high salinities,
The N-acetylation of amino acids such as ornithine and along with proline, but the ectoine to proline ratio strongly
lysine converts positively charged amino acids into neutral increases in the late stationary phase of the cell cycle [51].
zwitterionic (i.e., dipolar) molecules. The role of Nδ-acetyl- The ectABC genes, coding for the enzymes of the ectoine
ornithine in osmoadaptation was first confirmed in Bacillus biosynthetic pathway, form an operon that is expressed in a
strains. This solute was later detected at low levels in almost salinity-dependent manner, with maximal expression at 3 M
all the Bacillus species investigated as well as in Plano- NaCl.
COMPATIBLE SOLUTES IN PROKARYOTES
INT. MICROBIOL. Vol.11, 2008 155

philus, which belongs to a very ancient lineage of the


Trehalose and sucrose: the wide- Actinobacteria and is one of the most radiation-resistant
spread sugars organisms known [2,8; our unpublished results]. Rubro-
bacter xylanophilus accumulates high levels of trehalose as
Trehalose is a nonreducing glucose disaccharide that occurs the major organic solute under all conditions tested, includ-
in a wide variety of organisms, from Bacteria and Archaea to ing those for optimal growth [7]. However, functional char-
fungi, plants, and invertebrates. It protects numerous biolog- acterization of these pathways is required to ascertain their
ical structures against various kinds of stress, including des- independent roles in trehalose metabolism. Nonetheless, the
iccation, oxidation, heat, cold, dehydration, and hyperosmot- importance of pathway multiplicity and the ubiquity of tre-
ic conditions. In addition, trehalose is a source of carbon and halose in this radiation-resistant thermophile foreshadow an
energy and a signaling molecule in specific metabolic path- essential role in R. xylanophilus physiology.
ways [5]. To date, five different enzymatic systems have been Sucrose is a non-reducing disaccharide of glucose and
described for trehalose synthesis: the TPS/TPP, TreS, TreY- fructose that is widely distributed in plants [28]. In prokary-
TreZ, TreP, and TreT enzymes (Fig. 2) [1]. The properties of otes, however, only freshwater and marine cyanobacteria as
TPS/TPP, TreS, TreY-TreZ, and TreT have been reviewed by well as some proteobacteria are known to accumulate it. In
Empadinhas and da Costa [6]. Most microorganisms rely on these bacteria sucrose behaves as a compatible solute in
a single pathway, but some, including Mycobacterium tuber- osmotic stress. The pathways for the synthesis of sucrose
culosis, Corynebacterium glutamicum, and T. termophilus, were first characterized in higher plants and later in green
have two or even three pathways [1,57]. It must be noted that algae and in cyanobacteria [29]. One pathway involves two
the genes for four pathways for the synthesis of trehalose, steps catalyzed by sucrose-6-phosphate synthase (SPS) and
i.e., those encoding TPS/TPP, TreS, TreY/TreZ, and TreT, sucrose-6-phosphate phosphatase (SPP) through a phospho-
have been identified in the thermophile Rubrobacter xylano- rylated intermediate, much like the TPS/TPP pathway for tre-

Fig. 2. Pathways for the synthesis of tre-


halose. TPS, trehalose-6-phosphate syn-
thase; TPP, trehalose-6-phosphate phos-
phatase; TreS, trehalose synthase; TreY,
Int. Microbiol.

maltooligosyltrehalose synthase; TreZ,


maltooligosyltrehalose trehalohydrolase;
TreP, trehalose phosphorylase; TreT, tre-
halose glycosyltransferring synthase.
EMPADINHAS, DA COSTA
156 INT. MICROBIOL. Vol. 11, 2008

halose. An alternative pathway, which was found in higher centrations, to the osmolyte pool of hyperthermophilic
plants and in some filamentous cyanobacteria of the genus organisms. It is worth mentioning here that DIP is the major
Anabaena, uses a sucrose synthase (SuS) that catalyzes the organic osmolyte in Pyrolobus fumarii, which seems to be
condensation of ADP-glucose and fructose into sucrose [3]. the most thermophilic of all Archaea known [11].
However, the activity of SuS is reversible, which indicates The biosynthetic pathway for DIP and the key genes
the potential involvement of this enzyme in sucrose catabo- involved was elucidated by Rodionov and collaborators [44]
lism. The complete genome sequences of many cyanobacte- and Rodrigues and collaborators [45]. The genes for CTP:L-
ria and proteobacteria revealed additional sps, spp, and sus myo-inositol-1-phosphate cytidylyltransferase and for DIPP
genes [47]. The sps and spp genes are located in separate synthase [catalyzing the synthesis of di-myo-inositol-1,3-
regions of the chromosome, either organized in an operon- phosphate-1-phosphate (DIPP), the phosphorylated precursor
like structure or fused into a bifunctional gene. A unique of DIP], have been identified in several hyperthermophiles
genomic organization has been detected in the proteobacteri- and also in Rubrobacter xylanophilus, organisms known to
um Nitrosomonas europaea, in which genes for the two accumulate DIP and for which genome sequences are avail-
sucrose pathways are combined in an operon-like structure able [45]. The DIPP synthase activity is part of a bifunction-
[29]. Two forms of SPS from Anabaena and Synechocystis al enzyme that catalyzes the condensation of CTP and L-myo-
strains have been characterized [40]. Although both enzymes inositol-1-phosphate into CDP-L-myo-inositol, and of the lat-
are active, the reasons underlying this duplication remain ter two into DIPP.
unknown. In contrast with the SPS from plants, the enzymes Other polyol-phosphodiesters have been detected in
show low specificity for NDP-glucose donors. hyperthermophilic organisms. Di-mannosyl-di-myo-inositol
phosphate (DMDIP) has been identified in members of the
genus Thermotoga, in which the concentration increases
Phosphodiesters: compatible solutes mostly in response to heat stress [30]. Diglycerol phosphate
in hyper/thermophilic prokaryotes (DGP) has been identified in members of the genus
Archaeoglobus, where it accumulates under salt stress (Fig.
Among microorganisms with optimum growth temperatures 3), and glyceryl-myo-inosityl phosphate (GIP), a structural
above 80ºC, di-myo-inositol phosphate (DIP) is a widespread chimera of DIP and DGP, has been identified in Aquifex
organic solute (Fig. 3) [31,48]. DIP was first identified in the pyrophilus and Archaeoglobus fulgidus (Fig. 3) [24,25,30].
archaeon Pyrococcus woesei and was later found to accumu- GIP might play a dual role in osmo- and thermoprotection,
late in hyperthermophiles of the genera Aeropyrum, Aquifex, since its intracellular levels increases primarily in response to
Archaeoglobus, Pyrodictium, Pyrolobus, Stetteria, Thermo- combined heat and osmotic stresses.
coccus, and Thermotoga, mostly in response to supra-optimal Rubrobacter xylanophilus accumulates the organic solute
growth temperatures [49,53]. For this reason DIP was con- di-N-acetyl-glucosamine phosphate (DAGAP), whose struc-
sidered to play a role in protecting against the effects of ture (Fig. 3) is similar to that of the phosphodiester compati-
extremely high temperatures. However, DIP has been also ble solutes found in hyperthermophiles, DIP, DGP, GIP, and
reported to accumulate in Rubrobacter xylanophilus (optimal DMDIP. However, the role of DAGAP as a compatible solute
growth at 60ºC), mostly at supra-optimal growth tempera- has been refuted because its concentrations are always too
tures; it is the first report of the occurrence of this solute in low to contribute to the cell’s osmotic balance. It must be
an organism with an optimum growth temperature consider- noted that all phosphodiester solutes from hyperthermophiles
ably below 80ºC [7]. Later, DIP was also reported to occur in reported to date are polyol derivatives, whereas DAGAP has
the thermophilic bacterium Persephonella marina [49]. a phosphate group linking two sugar moieties [7].
Since many lipids of Archaea and Bacteria lack myo-
inositol-phosphate in the polar head groups, it was suggested
that DIP is a byproduct of the synthesis of inositol-containing Cyclic-bisphosphoglycerate in metha-
phospholipids. In fact, inositol-containing phospholipids are nogenic archaea
very common in Bacteria, abundant in crenarchaeotes, and
can reach high levels also in some euryarchaeotes, namely in Compatible solutes of Archaea usually resemble their bacte-
Pyrococcus and Thermococcus. Although DIP and its precur- rial counterparts, except that most of them have a negative
sors could serve dual functions, the compound must be charge due to the addition of carboxylate, phosphate, or sul-
viewed as a de facto compatible solute that counterbalances fate groups [4,46]. The usual negative charge of archaeal
the positive charge of K+ and contributes, by virtue of its con- compatible solutes is believed to neutralize the positive charge
COMPATIBLE SOLUTES IN PROKARYOTES
INT. MICROBIOL. Vol.11, 2008 157

are also present in the genomes of hyperthermophilic archaea


of the genus Pyrococcus, although cBPG has never been
detected in these organisms. Remarkably, these genes are
also found in the genome of Rubrobacter xylanophilus
although cBPG has never been detected.

Mannosylglycerate and mannosylglyc-


eramide

Two thermophilic bacteria, Thermus thermophilus and


Rhodothermus marinus, contain mannosylglycerate (MG),
which accumulates in response to salt stress (Fig. 4) [36].
This compound has been detected also in many hyperther-
mophilic archaea, where it likewise accumulates concomi-
tantly with increasing salinity of the medium [48]. This
organic solute, first discovered in the red alga Polysiphonia
fastigiata, is found in several members of the order
Ceramiales and used to be considered a taxonomic marker
for this order [20]. However, members of the orders
Gelidiales and Gigartinales also accumulate MG and its tax-
onomic value has been lost [20].
The concentrations of MG in red algae do not usually cor-
relate with increases in salinity; therefore, its role as a true
compatible solute in these organisms remains an open ques-
tion. Although present in many red algae, the apparent
restriction of MG to thermophilic bacteria and hyperther-
mophilic archaea led to the hypothesis that MG plays a major
role in thermal adaptation. However, DIP has been consid-
ered the dominant intracellular organic solute in hyperther-
mophiles cultured at supra-optimal temperatures [49]. It must
be noted that the hyperthermophile Palaeococcus ferrophilus
does not accumulate DIP and the bacterium has been found
Int. Microbiol.

to accumulate increasing levels of MG under salt or thermal


stresses alike [35].
Strains of the thermophilic and moderately halotolerant
Fig. 3. Structure of known phosphodiester compatible solutes, primarily bacterium Thermus thermophilus can be divided in three
found in hyperthermophilic organisms. groups according to their salt tolerance: some can grow at
salinities as high as 5–6% NaCl and have been found to pos-
sess genes for the synthesis of MG and trehalose, the domi-
of potassium. Cyclic-2,3-bisphosphoglycerate (cBPG) is one nant compatible solute. Strains that cannot grow above 3%
of those compatible solutes and, to date, it has been detected NaCl have functional genes for the synthesis of MG but can-
only in methanogenic archaea with a broad range of optimum not synthesize trehalose. Interestingly, one strain lacks the
growth temperatures [12,54,55]. The role of cBPG in genes for MG synthesis but contains genes for trehalose
osmoadaptation is still unclear but alternative roles have been biosynthesis and cannot grow when the salinity of the medi-
suggested for this solute, namely, in thermal protection, as an um exceeds 1% NaCl [6]. These data suggest that MG is
intermediate in a gluconeogenic pathway, and as a phosphate required for low-level salt adaptation and that it might some-
reservoir for ATP synthesis [12,55]. The genes for the synthe- how be involved in the switch of compatible solute pools to
sis of cBPG have been identified in Methanothermus fervidus trehalose. This is similar to the osmoadaptation phenomena
as well as in several methanogenic archaea [26]. These genes reported in Halobacillus halophilus, which switches its
EMPADINHAS, DA COSTA
158 INT. MICROBIOL. Vol. 11, 2008

Int. Microbiol.
Fig. 4. Structure of glucosylglycerate and of the naturally occurring derivatives glucosyl-glucosylglycerate and mannosyl-glucosyl-
glycerate.

osmolyte pool from glutamate to proline when the salinity Erwinia chrysanthemi under combined salt stress and nitro-
increases. It turns out that glutamate is itself involved in the gen-limiting conditions, replacing glutamate and glutamine,
up-regulation of proline biosynthetic genes [51]. Manno- the compatible solutes when abundant sources of nitrogen are
sylglycerate serves as a compatible solute in Rhodothermus present in the medium [13]. Glucosylglycerate appeared to
marinus, increasing with the salinity of the medium. be a rare organic solute with a restricted distribution among
However, at salinities close to the maximum for the growth mesophilic bacteria until it was unexpectedly identified in the
of this bacterium, a neutral form of MG, designated manno- thermophilic bacterium Persephonella marina, a member of
sylglyceramide (MGA), becomes the dominant compatible the Aquificales, where it was suggested to act as a true com-
solute [56]. It is possible that MG is the substrate for amida- patible solute under salt stress [49]. This observation argued
tion, but the corresponding gene and enzyme have not been against the initial hypothesis of a restricted role for GG in
identified. In this organism, MG also has been implicated in organisms living at low temperatures and suggests that GG
the response of R. marinus to thermal stress [6]. Manno- and MG are functionally interchangeable in the adaptation to
sylglycerate also accumulates in Rubrobacter xylanophilus, stress.
which represents a deep-branching lineage of the phylum A GG-derivative compatible solute, glucosyl-(1,6)-gluco-
Actinobacteria [7]. However, the compound is constitutively sylglycerate (GGG) (Fig. 4), has been detected in Perse-
accumulated and neither salt, thermal stress, nor the medium phonella marina [49]. However, detailed information on the
composition has major effects on the intracellular levels of conditions leading to GG and GGG accumulation is not
this solute. available. This compound was previously detected in trace
amounts in mycobacteria, where it was considered an inter-
mediate in the synthesis of a rare methylglucose lipopolysac-
Glucosylglycerate, glucosyl-glucosyl- charide [18].
glycerate, and mannosyl-glucosylglyc- Finally, an additional and unique compatible solute
erate derived from GG has been identified in Petrotoga miotherma
and characterized as mannosyl-(1,2)-glucosylglycerate
Glucosylglycerate (GG) (Fig. 4) is structurally analogous to (MGG) (Fig. 4) [17]. The levels of MGG increase with the
MG and was originally identified in the marine cyanobacteri- NaCl concentration of the medium up to the optimum for
um Agmenellum quadruplicatum when grown under nitro- growth, being replaced by proline and α-glutamate at higher
gen-limiting conditions. This compound was recently shown NaCl concentrations. Curiously, the strategy for MGG syn-
to behave as a compatible solute in the g-proteobacterium thesis does not appear to resemble that of R. marinus, in
COMPATIBLE SOLUTES IN PROKARYOTES
INT. MICROBIOL. Vol.11, 2008 159

Int. Microbiol.
Fig. 5. Taxonomical distribution of the organisms (genera) in which glucosylglycerate (solid black) and mannosylglycerate (solid
gray) have been detected. Open boxes represent organisms containing genes for the synthesis of glucosylglycerate (black line)
or mannosylglycerate (gray line).

which a pre-existing compatible solute (MG) is converted patible solute, and P. putida accumulates mannitol when
into a different form (MGA). In fact, GG has not been detect- exposed to osmotic stress. The accumulation of a polyol by
ed in P. miotherma under any of the conditions tested and the Z. mobilis seems to reflect convergent evolution for osmoad-
precursor for MGG biosynthesis has been identified to be aptation. In fact, this organism, like many yeasts, has devel-
GPG (our unpublished results). Figure 5 shows the taxonom- oped a strategy to modify a readily available and abundant
ical distribution of the organisms (genera) in which glucosyl- sugar into a compatible solute [27].
glycerate and mannosylglycerate have been detected. Cyanobacteria with moderate salt tolerance synthesize glu-
cosylglycerol and use it as a compatible solute in salt stress.
This rare compatible solute is a polyol derivative structurally
Polyols: rarely used as compatible related to galactosylglycerol, found in some red algae [19]. The
solutes in prokaryotes halotolerant proteobacteria Pseudomonas mendocina and
Stenotrophomonas rhizophila also produce glucosylglycerol
The polyols glycerol, arabitol, sorbitol, mannitol and inositol [33,43]. The cyanobacterium Synechocystis sp. PCC6803 pro-
are archetypal compatible solutes of halotolerant fungi and of duces glucosylglycerol mainly by de novo synthesis or, prefer-
some algae and plants [14]. Bacteria rarely use polyols as entially, takes it up if available in the environment [32]. A
compatible solutes, with the few exceptions including remarkable strategy of compatible solute accumulation in
Zymomonas mobilis and Pseudomonas putida [22,27]. Archaea involves the utilization of polyol phosphodiesters,
Unlike most heterotrophic bacteria, Z. mobilis can convert namely diglycerol phosphate and di-myo-inositol-phosphate, as
sucrose into glucose and sorbitol, using the latter as a com- mentioned earlier in this review [48,49].
EMPADINHAS, DA COSTA
160 INT. MICROBIOL. Vol. 11, 2008

5. Elbein AD, Pan YT, Pastuszak I, Carroll D (2003) New insight on tre-
halose: a multifunctional molecule. Glycobiology 13:17R-27R
Concluding remarks 6. Empadinhas N, da Costa MS (2006). Diversity and biosynthesis of com-
patible solutes in hyper/thermophiles. Int Microbiol 9:199-206
In the prokaryotic world, the use of small organic molecules, 7. Empadinhas N, Mendes V, Simoes C, Santos MS, Mingote A, Lamosa
P, Santos H, da Costa MS (2007) Organic solutes in Rubrobacter xyla-
such as amino acids, sugars or their derivatives, to cope with nophilus: the first example of di-myo-inositol-phosphate in a ther-
unfavorable environmental conditions is a widely dissemi- mophile. Extremophiles 11:667-673
nated strategy. Some of these molecules, including trehalose, 8. Ferreira AC, Nobre MF, Moore E, Rainey FA, Battista JR, da Costa MS
(1999) Characterization and radiation resistance of new isolates of
might have evolved early in the history of life, as reflected Rubrobacter radiotolerans and Rubrobacter xylanophilus. Extremo-
both by the plethora of different biosynthetic pathways and philes 3:235-238
by their ubiquity, from the lowest to the higher branches of 9. Galinski EA, Pfeiffer HP, Trüper HG (1985) 1,4,5,6-Tetrahydro-2-
methyl-4-pyrimidinecarboxylic acid. A novel cyclic amino acid from
the tree of life, whether as a vertically inherited trend or hor-
halophilic phototrophic bacteria of the genus Ectothiorhodospira. Eur J
izontally acquired [15,52]. Other such molecules, such as Biochem 149:135–139
DIP or cBPG, which have a considerably narrower distribu- 10. Galinski EA (1995) Osmoadaptation in bacteria. Adv Microb Physiol
tion, seem to have been neglected by the evolutionary cool- 37:272–328.
11. Gonçalves LG, Lamosa P, Huber R, Santos H (2008) Di-myo-inositol
ing that probably followed the early burst of life on Earth. phosphate and novel UDP-sugars accumulate in the extreme hyperther-
Compatible solutes including the related MG and GG, which mophile Pyrolobus fumarii. Extremophiles 12:383-389
have found their way from hyper/thermophilic Bacteria and 12. Gorkovenko A, Roberts MF (1993) Cyclic 2,3-diphosphoglycerate as a
component of a new branch in gluconeogenesis in Methanobacterium
Archaea to red algae and possibly fungi and mosses, and for thermoautotrophicum ΔH. J Bacteriol 175:4087-4095
which evolution has found more than one pathway, continue 13. Goude R, Renaud S, Bonnassie S, Bernard T, Blanco C (2004)
to inspire and challenge biologists since despite their func- Glutamine, glutamate, and α-glucosylglycerate are the major osmotic
solutes accumulated by Erwinia chrysanthemi strain 3937. Appl
tional interplay they are rarely found in the same phylogenet-
Environ Microbiol 70:6535-6541
ic clusters. Additional roles for these molecules no doubt will 14. Grant WD (2004) Life at low water activity. Philos Trans R Soc Lond B
be soon unveiled—not only structural roles like those played Biol Sci 359:1249-1266
by trehalose and GG in mycobacteria or Nocardia, but also 15. Guerrero R, Berlanga M (2006) Life’s unity and flexibility: the ecolog-
ical link. Int Microbiol 9:225-235
functional ones at the physiological and molecular levels. 16. Hagemann M, Effmert U, Kerstan T, Schoor A, Erdmann N (2001)
The study of the genes and synthetic pathways of compatible Biochemical characterization of glucosylglycerol-phosphate synthase of
solutes, the regulation of the biosynthesis of these com- Synechocystis sp. strain PCC 6803: comparison of crude, purified, and
recombinant enzymes. Curr Microbiol 43:278-283
pounds, and of the phylogenetic relationship among their nat- 17. Jorge CD, Lamosa P, Santos H (2007) α–D–Mannopyran-
ural producers will continue to expand our knowledge on the osyl–(1→2)–α–D–glucopyranosyl–(1→2)–glycerate in the ther-
evolution of prokaryotic adaptations to stress. mophilic bacterium Petrotoga miotherma–Structure, cellular content
and function. FEBS J 274:3120-3127
Acknowledgements. We are grateful to the many collaborators and 18. Kamisango K, Dell A, Ballou CE (1987) Biosynthesis of the mycobac-
co-authors cited, all of whom decisively contributed to our knowledge of terial o-methylglucose lipopolysaccharide. Characterization of putative
compatible solutes biology. We thank Helena Santos and her group (ITQB, intermediates in the initiation, elongation, and termination reactions. J
Oeiras, Portugal) for helpful discussions and valuable collaboration. Part of Biol Chem 262:4580-4586
the work was supported by Fundação para a Ciência e a Tecnologia, FCT, 19. Karsten U, West JA, Zuccarello GC, Engbrodt R, Yokoyama A, Hara Y,
Portugal, and FEDER, Projects POCI/BIA-MIC/56511/2004 and A005/2005 Brodie J (2003) Low molecular weight carbohydrates of the
Action V.5.1. Bangiophycidae (Rhodophyta). J Phycol 39:584-589
20. Karsten U, Gors S, Eggert A, West JA (2007) Trehalose, digeneaside,
and floridoside in the Florideophyceae (Rhodophyta)–a reevaluation of
its chemotaxonomic value. Phycologia 46:143-150
References 21. Kempf B, Bremer E (1998) Uptake and synthesis of compatible solutes
as microbial stress responses to high-osmolality environments Arch
1. Avonce N, Mendoza-Vargas A, Morett E, Iturriaga G (2006) Insights on Microbiol 170:319-330
the evolution of trehalose biosynthesis. BMC Evol Biol 6:109 22. Kets EP, Galinski EA, de Wit M, de Bont JA, Heipieper HJ (1996)
2. Carreto L, Moore E, Nobre MF, Wait R, Riley PW, Sharp RJ, da Costa Mannitol, a novel bacterial compatible solute in Pseudomonas putida
MS (1996) Rubrobacter xylanophilus sp. nov., a new thermophilic S12. J Bacteriol 178:6665-6670
species isolated from a thermally polluted effluent. Int J Syst Bacteriol 23. Killham K, Firestone MK (1984) Salt stress control of intracellular
46:460-465 solutes in streptomycetes indigenous to saline soils. Appl Environ
3. Curatti L, Porchia AC, Herrera-Estrella L, Salerno GL (2000) A Microbiol 47:301-306
prokaryotic sucrose synthase gene (susA) isolated from a filamentous 24. Lamosa P, Burke A, Peist R, et al. (2000) Thermostabilization of pro-
nitrogen-fixing cyanobacterium encodes a protein similar to those of teins by diglycerol phosphate, a new compatible solute from the
plants. Planta 211:729-735 hyperthermophile Archaeoglobus fulgidus. Appl Environ Microbiol
4. da Costa MS, Santos H, Galinski EA (1998) An overview of the role and 66:1974-1979
diversity of compatible solutes in Bacteria and Archaea. Adv Biochem 25. Lamosa P, Gonçalves LG, Rodrigues MV, Martins LO, Raven ND,
Eng Biotechnol 61:117-153 Santos H (2006) Occurrence of 1-glyceryl-1-myo-inosityl phosphate in
COMPATIBLE SOLUTES IN PROKARYOTES
INT. MICROBIOL. Vol.11, 2008 161

hyperthermophiles. Appl Environ Microbiol 72:6169-6173 43. Roder A, Hoffmann E, Hagemann M, Berg G (2005) Synthesis of the
26. Lehmacher A, Vogt AB, Hensel R (1990) Biosynthesis of cyclic 2,3- compatible solutes glucosylglycerol and trehalose by salt-stressed cells
diphosphoglycerate. Isolation and characterization of 2-phosphoglycer- of Stenotrophomonas strains. FEMS Microbiol Lett 243:219-226
ate kinase and cyclic 2,3-diphosphoglycerate synthetase from 44. Rodionov DA, Kurnasov OV, Stec B, Wang Y, Roberts MF, Osterman
Methanothermus fervidus. FEBS Lett 272:94-98 AL (2007) Genomic identification and in vitro reconstitution of a com-
27. Loos H, Kramer R, Sahm H, Sprenger GA (1994) Sorbitol promotes plete biosynthetic pathway for the osmolyte di-myo-inositol-phosphate.
growth of Zymomonas mobilis in environments with high concentra- Proc Natl Acad Sci USA 104:4279-4284
tions of sugar: evidence for a physiological function of glucose-fructose 45. Rodrigues MV, Borges N, Henriques M, et al. (2007) Bifunctional
oxidoreductase in osmoprotection. J Bacteriol 176:7688-7693 CTP:inositol-1-phosphate cytidylyltransferase/CDP-inositol:inositol-1-
28. Lunn JE, MacRae E (2003) New complexities in the synthesis of phosphate transferase, the key enzyme for di-myo-inositol-phosphate
sucrose. Curr Opin Plant Biol 6:208-214 synthesis in several (hyper)thermophiles. J Bacteriol 189:5405-5412
29. Lunn JE (2002) Evolution of sucrose synthesis. Plant Physiol 46. Roesser M, Muller V (2001) Osmoadaptation in bacteria and archaea:
128:1490-500 common principles and differences. Environ Microbiol 3:743-754
30. Martins LO, Carreto LS, da Costa MS, Santos H (1996) New compati- 47. Salerno GL, Curatti L (2003) Origin of sucrose metabolism in higher
ble solutes related to di-myo-inositol-phosphate in members of the order plants: when, how and why? Trends Plant Sci 8:63-69
Thermotogales. J Bacteriol 178:5644-5651 48. Santos H, da Costa MS (2002) Compatible solutes of organisms that live
31. Martins LO, Huber R, Huber H, Stetter KO, da Costa MS, Santos H in hot saline environments. Environ Microbiol 4:501-509
(1997) Organic solutes in hyperthermophilic Archaea. Appl Environ 49. Santos H, Lamosa P, Borges N, Faria TQ, Neves C (2007) The physio-
Microbiol 63:896-902 logical role, biosynthesis and mode of action of compatible solutes from
32. Mikkat S, Hagemann M, Schoor A (1996) Active transport of glucosyl- (hyper)thermophiles, In: Gerday C, Glandorff N (eds.) Physiology and
glycerol is involved in salt adaptation of the cyanobacterium biochemistry of extremophiles, ASM Press, Washington, DC, pp 86-103
Synechocystis sp. strain PCC 6803. Microbiology 142:1725-732 50. Saum SH, Sydow JF, Palm P, Pfeiffer F, Oesterhelt D, Müller V (2006)
33. Mikkat S, Galinski EA, Berg G, Minkwitz A, Schoor A (2000) Salt adap- Biochemical and molecular characterization of the biosynthesis of glut-
tation in pseudomonads: characterization of glucosylglycerol-synthesiz- amine and glutamate, two major compatible solutes in the moderately
ing isolates from brackish coastal waters and the rhizosphere. Syst Appl halophilic bacterium Halobacillus halophilus. J Bacteriol 188:6808-
Microbiol 23:31-40 6815
34. Müller V, Oren A (2003) Metabolism of chloride in halophilic prokary- 51. Saum SH, Müller V (2008) Growth phase-dependent switch in osmolyte
otes. Extremophiles 7:261-266 strategy in a moderate halophile: ectoine is a minor osmolyte but major
35. Neves C, da Costa MS, Santos H (2005) Compatible solutes of the stationary phase solute in Halobacillus halophilus. Environ Microbiol
hyperthermophile Palaeococcus ferrophilus: osmoadaptation and ther- 10:716-726
moadaptation in the order thermococcales. Appl Environ Microbiol 52. Schmidt T (2006) The maturing of microbial ecology. Int Microbiol
71:8091-8098 9:217-223
36. Nunes OC, Manaia CM, da Costa MS, Santos H (1995) Compatible 53. Scholz S, Sonnenbichler J, Schäfer W, Hensel R (1992) Di-myo-inosi-
solutes in the thermophilic bacteria Rhodothermus marinus and tol-1,1′-phosphate: a new inositol phosphate isolated from Pyrococcus
“Thermus thermophilus”. Appl Environ Microbiol 61:2351-2357 woesei. FEBS Lett 306:239-242
37. Oren A (2008) Microbial life at high salt concentrations: phylogenetic 54. Seely RJ, Fahrney DE (1983) A novel diphospho-P,P′-diester from
and metabolic diversity. Saline Systems 4:2 Methanobacterium thermoautotrophicum. J Biol Chem 258:10835-
38. Pflüger K, Baumann S, Gottschalk G, Lin W, Santos H, Müller V (2003) 10838
Lysine-2,3-aminomutase and β-lysine acetyltransferase genes of 55. Shima S, Herault DA, Berkessel A, Thauer RK (1998) Activation and
methanogenic archaea are salt induced and are essential for the biosyn- thermostabilization effects of cyclic 2,3-diphosphoglycerate on
thesis of Nε-acetyl-β-lysine and growth at high salinity. Appl Environ enzymes from the hyperthermophilic Methanopyrus kandleri. Arch
Microbiol 69:6047-55 Microbiol 170:469-472
39. Poolman B, Glaasker E (1998) Regulation of compatible solute accu- 56. Silva Z, Borges N, Martins LO, Wait R, da Costa MS, Santos H (1999)
mulation in bacteria. Mol Microbiol 29:397-407 Combined effect of the growth temperature and salinity of the medium
40. Porchia AC, Salerno GL (1996) Sucrose biosynthesis in a prokaryotic on the accumulation of compatible solutes by Rhodothermus marinus
organism: Presence of two sucrose-phosphate synthases in Anabaena and Rhodothermus obamensis. Extremophiles 3:163-172
with remarkable differences compared with the plant enzymes. Proc 57. Silva Z, Alarico S, Nobre A, Horlacher R, Marugg J, Boos W, Mingote
Natl Acad Sci USA 93:13600-13604 AI, da Costa MS (2003) Osmotic adaptation of Thermus thermophilus
41. Roberts MF, Lai MC, Gunsalus RP (1992) Biosynthetic pathways of the RQ-1: a lesson from a mutant deficient in the synthesis of trehalose. J
osmolytes Nε-acetyl-β-lysine, β-glutamine, and betaine in Bacteriol 185:5943-5952
Methanohalophilus strain FDF1 suggested by nuclear magnetic reso- 58. Wohlfarth A, Severin J, Galinski EA (1993) Identification of N?-acety-
nance analyses. J Bacteriol 174:6688-6693 lornithine as a novel osmolyte in some Gram-positive halophilic eubac-
42. Robertson DE, Roberts MF, Belay N, Stetter KO, Boone DR (1990) teria. Appl Microbiol Biotechnol 39:568-573
Occurrence of β-glutamate, a novel osmolyte, in marine methanogenic
bacteria. Appl Environ Microbiol 56:1504-1508

View publication stats

You might also like