A THESIS ABOUT BIOSURFACTANT

SURFACTANT

The word “ surfactant” is a contraction of three word “ Surface Active Agent”.

So surfactants are materials that lower the surface tension or interfacial
tension between two liquids or between a liquid and solid . In the general
sense , any material that affects the interfacial surface tension , can be
consider a surfactant ,but in the practical sense, surfactants may act as
wetting agents, emulsifiers, foaming agents, and dispersants.

Surface active agents play an important role as cleaning, wetting, dispersing,

emulsifying, foaming and antifoaming agents in many practical applications
and products, including: paints, emulsions adhesives, inks, biocides,
shampoos, toothpastes, firefighting, detergents, insecticides, deinking of
recycled papers, ski waxes, spermicides .

In the bulk aqueous phase surfactants form masses, such as micelles where
the hydrophobic tails form the core and the hydrophilic heads are immersed
in the surrounding liquid . Others types of structures can also be formed, such
as spherical micelles or lipid bilayers . The shape of the molecules depends
upon the balance in size between hydrophilic head and hydrophobic tail. A
measure of this is the HLB, Hydrophilic-lipophilic balance. Higher HLB
surfactants (>10) are hydrophilic (“water loving”) and form O/W (Oil-in-water)
emulsions. Lipophilic surfactants possess low HLB values (1-10) and form W/O
(water-in-oil) emulsions. Dish detergents, surfactants for emulsion
polymerization, and the following example (SLS = Sodium Lauryl Sulfate) are
high HLB surfactants.

The dynamics of surface active agent adsorption is of great importance for

practical applications such as in emulsifying or coating processes as well as
foaming, where bubbles or drops are rapidly generated and need to be
stabilized.

As the interface is created, the adsorption is limited by the diffusion of the

surfactant to the interface, which can result in the kinetics being limited.
These energy barriers can be due to steric or electrostatic repulsions; steric
repulsions form the basis of how dispersants function. Surface rheology of
surfactant layers, are important to the stability of foams and emulsions.

Most surfactants’ “tails” are fairly similar, consisting of a hydrocarbon chain,

which can be branched, linear, or aromatic. Fluorosurfactants have
fluorocarbon chains. Siloxane surfactants have siloxane chains. Recent
advances in surfactant technology has seen the development of mixed chains
or/and complex structures.

SURFACTANT AND AMPHIPHILES

The word amphiphile was coined by Paul Winsor 50 years ago. It comes from
two Greek roots. First the prefix amphi which means "double", "from both
sides", "around", as in amphitheater or amphibian. Then the root philos
which expresses friendship or affinity, as in "philanthropist" (the friend of
man), "hydrophilic" (compatible with water), or "philosopher" (the friend of
wisdom or science).
An amphiphilic substance exhibits a double affinity, which can be defined
from the physico-chemical point of view as a polar-apolar duality. A typical
amphiphilic molecule consists of two parts: on the one hand a polar group
which contents heteroatoms such as O, S, P, or N, included in functional
groups such as alcohol, thiol, ether, ester, acid, sulfate, sulfonate, phosphate,
amine, amide etc… On the other hand, an essentially apolar group which is in
general an hydrocarbon chain of the alkyl or alkylbenzene type, sometimes
with halogen atoms and even a few nonionized oxygen atoms.

The polar portion exhibits an strong affinity for polar solvents, particularly
water, and it is often called hydrophilic part or hydrophile. The apolar part is
called hydrophobe or lipophile, from Greek roots phobos (fear) and lipos
(grease). The following formula shows an amphiphilic molecule which is
commonly used in shapoos (sodium dodecyl sulfate).

Because of its dual affinity, an amphiphilic molecule does not feel "at ease" in
any solvent, be it polar or non polar, since there is always one of the groups
which "does not like" the solvent environment. This is why amphiphilic
molecules exhibit a very strong tendency to migrate to interfaces or surfaces
and to orientate so that the polar group lies in water and the apolar group is
placed out of it, and eventually in oil.

In the following the word surface will be used to designate the limit between
a condensed phase and a gas phase, whereas the term interface will be used
for the boundary between two condensed phases. This distinction is handy
though not necessary, and the two words are often used indifferently
particularly in american terminology.

In English the term surfactant (short for surface-active-agent) designates a

substance which exhibits some superficial o interfacial activity. It is worth
remarking that all amhiphiles do not display such activity; in effect, only the
amphiphiles with more or less equilibrated hydrophilic and lipophilic
tendencies are likely to migrate to the surface or interface. It does not happen
if the amphiphilic molecule is too hydrophilic or too hydrophobic, in which
case it stays in one of the phases.

Amphiphiles exhibit other properties than tension lowering and this is why
they are often labeled according to their main use such as: soap, detergent,
wetting agent, disperssant, emulsifier, foaming agent, bactericide, corrosion
inhibitor, antistatic agent, etc… In some cases they are konwn from the name
of the structure they are able to build, i.e. membrane, microemulsion, liquid
crystal, liposome, vesicle or gel.

CLASIFICATION OF SURFACTANT

From the commercial point of view surfactants are often classified according
to their use. However, this is not very useful because many surfactants have
several uses, and confusions may arise from that. The most acepted and
scientifically sound classification of surfactants is based on their dissociation
in water. There are 4 types of surfactants with a brief review of each as
follows. These classifications are based upon the composition of the polarity
of the head group: nonionic, anionic, cationic, amphoteric.
ANIONIC SURFACTANT - Anionic surfactants contain anionic functional groups
at their head, such as sulfonate, phosphate, sulfate and carboxylates. Alkyl
sulfates include ammonium lauryl sulfate, sodium lauryl and the related alkyl-
ether sulfates sodium laureth sulfate, also known as sodium lauryl ether
sulfate (SLES), and sodium myreth sulfate. These are the most common
surfactants and comprise the alkyl carboxylates (soaps), such as sodium
stearate. The stearates comprise >50% of the global usage of surfactants.

Many of these find utilization in emulsion polymerization. Other anionic

surfactants include dioctyl sodium sulfosuccinate (DOSS),
perfluorooctanesulfonate (PFOS), linear alkylbenzene sulfonates (LABs) and
perfluorobutanesulfonate, as well as alkyl-aryl ether phosphates. More
specialized species include sodium lauroyl sarcosinate and carboxylate-based
fluorosurfactants such as perfluorononanoate, perfluorooctanoate (PFOA or
PFO).

NONIONIC SURFACTANT - Nonionic Surfactants come as a close second with

about 45% of the overall industrial production. They do not ionize in aqueous
solution, because their hydrophilic group is of a non- dissociable type, such as
alcohol, phenol, ether, ester, or amide. A large proportion of these nonionic
surfactants are made hydrophilic by the presence of a polyethylene glycol
chain, obtained by the polycondensation of ethylene oxide. They are called
polyethoxylated nonionics. In the past decade glucoside (sugar based) head
groups, have been introduced in the market, because of their low toxicity. As
far as the lipophilic group is concerned, it is often of the alkyl or alkylbenzene
type, the former coming from fatty acids of natural origin. The
polycondensation of propylene oxide produce a polyether which (in oposition
to polyethylene oxide) is slightly hydrophobic. This polyether chain is used as
the lipophilic group in the so-called polyEO- polyPO block copolymers, which
are most often included in a different class, e.g. polymeric surfactants.

CATIONIC SURFACTANT - Cationic Surfactants are dissociated in water into an

amphiphilic cation and an anion, most often of the halogen type. A very large
proportion of this class corresponds to nitrogen compounds such as fatty
amine salts and quaternary ammoniums, with one or several long chain of the
alkyl type, often coming from natural fatty acids. These surfactants are in
general more expensive than anionics, because of a the high pressure
hydrogenation reaction to be carried out during their synthesis. As a
consequence, they are only used in two cases in which there is no cheaper
substitute, i.e. (1) as bactericide, (2) as positively charged substance which is
able to adsorb on negatively charged substrates to produce antistatic and
hydrophobant effect, often of great commercial importance such as in
corrosion inhibition.
Most of cationic surfactants find use as anti-microbials, anti-fungals, etc. in
HI&I (Benzalkonium chloride (BAC), Cetylpyridinium chloride (CPC),
Benzethonium chloride (BZT). The cationic nature of the surfactants is not
typically consistent with the world of non-ionic and anionic charges, and they
disrupt cell membranes of bacteria and viruses. Permanently charged
quaternary ammonium cations include: Alkyltrimethylammonium salts: cetyl
trimethylammonium bromide (CTAB) and cetyl trimethylammonium chloride
(CTAC).

ZWITTERIONIC SURFACTANT - Zwitterionic (amphoteric) surfactants have

both cationic and anionic centers attached to the same molecule. The anionic
part can be variable and include sulfonates, as in the sultaines CHAPS (3-[(3-
Cholamidopropyl)dimethylammonio]-1-propanesulfonate). Betaines such as
cocamidopropyl betaine have a carboxylate with the ammonium. The cationic
part is based on primary, secondary, or tertiary amines or quaternary
ammonium cations. Zwitterionic surfactants are often sensitive to pH and will
behave as anionic or cationic based on pH. Fast dry (“coacervation”) latex
traffic paints are based on this concept, with a drop in pH triggering the latex
in the paint to coagulate.

A few commonly used surfactans are given below

Some amphoteric surfactants are insensitive to pH, whereas others are
cationic at low pH and anionic at high pH, with an amphoteric behavior at
intermediate pH. Amphoteric surfactants are generally quite expensive, and
consequently, their use is limited to very special applications such as
cosmetics where their high biological compatibility and low toxicity is of
primary importance.

The past two decades have seen the introduction of a new class of surface
active substance, so-called polymeric surfactants or surface active polymers,
which result from the association of one or several macromolecular structures
exhibiting hydrophilic and lipophilic characters, either as separated blocks or
as grafts. They are now very commonly used in formulating products as
different as cosmetics, paints, foodstuffs, and petroleum production
additives.

PROPERTIES OF SURFACTANT

MICELLIZATION –

One of the most interesting properties of surfactants in solution is their ability

to self aggregate to form association colloids known as micelles, accompanied
by an overall decrease in the free energy of the system.2,4 At very low
concentration, the surfactant molecules are preferentially adsorbed at air
water interface with its hydrophobic tail pointing away from the water
surface thereby lowering the interfacial tension.3 As the concentration
increases, the adsorption at the air water interface becomes stronger forming
a condensed monolayer, known as Gibb’s monolayer at the interface after
which any further addition of surfactant molecules remain in the aqueous
phase.2,20 When the concentration of the surfactant molecules in the bulk of
the solution exceeds a limiting value, the surfactant molecules self aggregate
to form micelle which is manifested by an abrupt change in many physic-
chemical properties.

A schimetics representation of surfactant in solution is given below

The narrow concentration range over which these changes occur is known as
critical micelle concentration (CMC) and is perhaps the most important
characteristic property of a surfactant. The CMC may also be consideredas the
concentration at which micelles first appears in solution and is determined
from the marked changes in the plot of some physico-chemical properties of
the solution against the surfactant concentration.

Depending on the concentration of the surfactant, the geometric and

energetic factors, the size and shape of micelles fluctuate in a given system.3
Size of a micelle is expressed in terms of aggregation number i.e. the number
of monomer units present in a micelle. Generally, the aggregation number is
between 20 and 100 for single chain ionic surfactants while large aggregation
number of 1000 or more have been reported for non-ionic micelles especially
near the cloud point.3 The shape of micelles however may vary from spherical
to cylindrical, hexagonal, rod and to lamellar structure depending upon
various factors. Micelles are spontaneously formed and addition of more
surfactant leads to formation of more micelles increasing the micellar
concentration or the micellar growth while the surfactant monomers in the
system remain more or less unchanged.

The result is a decrease in the average distance between the micelles and
hence an increase in inter-micellar repulsion. In order to compensate it, the
spherical micelles may transform into worm like micelles thereby increasing
the distance between the micelles. The molecular architecture of a given
surfactant determines the type of aggregate into which a surfactant
associates in aqueous solution.

The spherical micelle proposed by Hartley2 is arguably the most successful

one for the purpose of rationalization of observed behaviour of micelles in
solution. In a typical spherical micelle, the hydrophobic tails of the surfactant
monomer in aqueous solution are preferentially associated to form the core
of the micelle while the hydrophilic heads are exposed to the water.
Immediate environment of the hydrophobic core that contains the
hydrophilic head along with the counterions constitute the stern layer, which
forms the inner portion of the electrical double layer surrounding the micelle.
 A schematic diagram of spherical micelle is given below

The main driving force behind the formation of surfactant aggregates in

aqueous solution is believed to be the Hydrophobic effect.30-32 The
hydrophobic effect promotes the aggregation of the surfactant molecules
while the electrostatic repulsion between the charged head groups opposes
it.33 Aggregation of hydrophobic groups in aqueous solution above a certain
concentration is due to the overlap of hydration shells formed around the
hydrophobic moieties.

Water undergoes a structural enhancement in the hydrophobic

hydration shells and upon aggregation, these shells overlaps and part of the
water molecules surrounding the individual solutes is released thereby de-
structuring the water structure. This accounts for the overall entropy gain
upon micellisation.
The tendency of a surfactant to form micelle in solution is largely dependent
on the type and nature of the surfactant.2,34 Surfactants with longer
hydrophobic tail (i.e. more hydrophobicity) generally exhibit greater tendency
towards micelles formation. With increase in the length of the hydrophobic
tail, the hydrophobic effect becomes stronger and consequently the CMC
decreases and larger micelles are formed. The aggregation number also
increases with hydrophobic chain length.35 Besides the chain length,
branching in the surfactant is also known to affect the CMC and the
aggregation number. The CMC of the branched surfactant has been shown to
be higher and the aggregation number lower than those of their linear chain.

Also micelles formation and and its stability depends on the solvent . The
polarity of solvent and its tendency to form hydrogen bond affects the
behaviour of micelles . Solvent like water,hydragene, formaldehyde, glycerol
which have greater tendency to form H-bond have prerequisite effect on
micellisation.

However micelle formation occur also in solvent which has less tendency or
no tendency to form H-bond like acetone, acetonitrile, dimethyl suphoxide
etc . In nonpolar solvent hydrophobic part increases which supresses the
tendency of surfactant molecule to form micelles.
BIOSURFACTANT –

Biosurfactants are diverse groups of surface active molecules or chemical

compounds synthesized by microorganisms. These amphiphilic compounds
are produced on living surfaces, mostly on microbial cell surfaces, or excreted
extracellulary.

Microbial suface active agents are a group of structurally diversed molecules

produced by different microorganisms and are mainly classified by their
chemical structure and their microbial origin. They are made up of a
hydrophilic moiety, comprising an acid, peptide cations, or anions, mono-, di-,
or polysaccharides, and a hydrophobic moiety of unsaturated or saturated
hydrocarbon chains or fatty acids. These structures confer a wide range of
properties, including the ability to lower surface and interfacial tension of
liquids and to form micelles and microemulsions between two different
phases. These compounds can be roughly divided into two main classes (Neu,
1996): low-molecular-weight compounds called biosurfactants, such as
lipopeptides, glycolipids, and proteins, and high-molecular-weight poly- mers
of polysaccharides, lipopolysaccharide proteins, or lipoproteins that are col-
lectively called bioemulsans (Rosenberg and Ron, 1997) or bioemulsifiers
(Smyth et al., 2010). The best studied microbial surfactants are glycolipids.
Among these, the best-known compounds are rhamnolipids, trehalolipids,
sophorolipids, and mannosylerythritol lipids (MELs). Rhamnolipid production
by Pseudomonas spe- cies has been extensively studied, and potential
applications have been proposed (Maier and Soberón-Chávez, 2000).
Rhamnolipids from Pseudomonas aerugi- nosa are currently commercialized
by Jeneil Biosurfactant, USA, mainly as a fun- gicide for agricultural purposes
or an additive to enhance bioremediation activities (Banat et al., 2010).

SURFACTANTS VS BIOSURFACTANTS –

Surfactants are a class of chemical compounds formed by molecular

structures with both hydrophilic and hydrophobic moieties that tend to be
distributed at the interface between liquid phases with different degree of
polarity. Most commercially available surfactants are synthesized from
petroleum derivatives which are hazardous to both environment and living
beings. Also petroleum products are non-renewable sourse which may be
finished in near future.

So considering above demerits of chemically synthesized surfactants and also

taking consideration into useful applications of surfactants, new types of
surfactants were developed which are synthesized from renewable sources
and also environmentally benign called as biosurfactants.

Most known biosurfactants are produced on substrates that are insoluble in

water, such as solid and liquid hydrocarbons, oils and fats, although many are
obtained from soluble substrates or a combination of the two (Van-Hamme et
al., 2006). The production of bio0surfactants with the use of renewable
substrates and different microbial species as well as the variation in culture
parameters (incubation time, stirring speed, pH of the medium and added
nutrients) allow the acquisition of compounds with distinct structural
characteristics and physical properties. This makes biosurfactants comparable
to or even better than synthetic surfactants in terms of efficiency, although
production costs do not yet allow greater competitiveness with their
petrochemical counterparts.

There are many advantages of biosurfactants when compared to their

chemically synthesized counterparts. Some of these are as follows:

• Biodegradability.

• Generally low toxicity.

• Biocompatibility and digestibility—which allows their application in cos-

metics and pharmaceuticals and as functional food additives.

• Availability of raw materials—BSs can be produced from inexpensive raw

materials that are available in large quantities; the carbon source may come
from hydrocarbons, carbohydrates, and/or lipids, which may be used sepa-
rately or in combination with each other.

• Acceptable production economics—depending upon the application, BSs

can also be produced from industrial wastes and by-products, and this is of
particular interest for bulk production (e.g., for use in petroleum-related
technologies).

• Use in environmental control—BSs can be efficiently used in handling

industrial emulsions, control of oil spills, biodegradation, and detoxification of
industrial effluents and in bioremediation of contaminated soil.

• Specificity—BSs, being complex organic molecules with specific func-

tional groups, are often specific in their action interest in detoxification of: de-
emulsification of industrial emulsions, specific cosmetic, pharmaceutical, and
food applications.

• Effectiveness—at extreme temperatures, pH, and salinity.

NEED OF SYNTHESIS OF BIO-SURFACTANTS-

There is no doubt that our lives have been enhanced by chemistry. All living
organisms are interlink with several chemicals and a chemistry behind the
chemicals. The chemical enterprise generates critical living needs such as food
for the world's population, achieves various medical wonders that save
millions of lives and improve people's health, and produces materials
essential to the present and future needs of mankind.

A line given by Clark in 1999 , ‘’ This role of chemistry is not generally

recognized by government or the public. In fact, chemicals, chemistry, and
chemists are actually seen by many as the cause of the problems"

The chemical industry releases more hazardous wastes to the environment

than any other industry sector . Basically petrolium industries where the use
of surfactant is maximum liberate hazardous chemicals.

Hydrophobic pollutants present in petroleum hydrocarbons and soil and

water environment require solubilization before being degraded by microbial
cells. Mineralization is governed by desorption of hydrocarbons from soil.
Surfactants can increase the surface area of hydrophobic materials, such as
pesticides in soil and water environment, thereby increasing their water
solubility. Hence, the presence of surfactants may increase microbial
degradation of pollutants. Use of biosurfactants (BSs) for degradation of
pesticides in soil and water environment has gained importance only recently.
BSs are attracting much interest due to their potential advantages over their
synthetic counterparts in many fields spanning environmental, food,
biomedical, and other industrial applications.

Also bio-surfactant is a part of green synthesis,a new methodology developed

by chemist to minimize the hazardous effect of chemically synthesize.

HISTORY OF BIOSURFACTANTS
RESEARCHES-
The development of green chemistry as a new science and BS research go
hand in hand. . In fact the roots of green chemistry go back to the 1950s,
when Henkel, a chemical company, started monitoring surfactant
concentrations in the Rhine River and developed the closed bottle test in
order to study the biodegradability of surfac- tants and since then has
strategically steered research and development according to environmental
principles, resulting in, for example, the development of zeolite A as an
alternative to phosphates in detergents (in order to avoid eutrophication of
sweet water lakes in Germany due to overfertilization). Development of
surfactants for carbon dioxide, enabling CO2 to be used as a solvent (e.g., in
dry cleaning), won the Presidential Green chemistry Challenge Awards.

The discovery of a new surfactant with high surface activity in supercritical

carbon dioxide opened a way to new processes in textile and metal industries
and for dry cleaning of clothes.

The production and use of BSs also imply to the principles of Green
Chemistry. Some are cited below:

• Olive oil mill effluent, a major pollutant of the agricultural industry in

Mediterranean countries, has been used as the raw material for rhamno- lipid
BS production by Pseudomonas sp. JAMM. The use of agroindustrial by-
products has been reported both for yeasts and bacteria (Makkar and
Cameotra, 2002). Sobrinho et al. (2008) used groundnut oil refinery residues
and corn steep liquor as substrates for anionic glycolipid production by
Candida sphaerica, while the biosynthesis of glycolipids by P. aeruginosa was
obtained using cashew apple juice as substrate (Rocha et al., 2007) and
vegetable oil refinery wastes (Raza et al., 2007), thus complying with the first
principle of green chemistry, that is, to Prevent Waste.

• A BS from P. aeruginosa was compared with a synthetic surfactant (Marlon

A-350) widely used in industry in terms of toxicity and mutagenic properties.
Both assays indicated the higher toxicity and mutagenic effect of the
chemical-derived surfactant, whereas BS was considered slightly to nontoxic
and nonmutagenic (Flasz et al., 1998). The comparison of acute and chronic
toxicity of three synthetic surfactants (Corexit, 9500, Triton X-100, and PSE-
61) and three microbial-derived surfactants (rhamnolipid, emulsan, and
biological cleanser PES-51) commonly used in oil spill remedia- tion revealed
that PES-61 (synthetic surfactant) and emulsan (BS) were the least toxic
whereas Triton X-100 (synthetic) was the most toxic (Edwards et al., 2003),
complying with principle 4 of green chemistry, that is, Design for Safer
Chemicals.

• Unlike synthetic surfactants, microbial-produced compounds are easily

degraded (Mohan et al., 2006) and particularly suited for environmental appli-
cations such as bioremediation (Deleu and Paquot, 2004; Mulligan, 2005). The
increasing environmental concern among consumers and the regulatory rules
imposed by governments forced industry to search for alternative products
such as BSs.

CLASSIFICATION OF BIOSURFACTANT-

Synthetic surfactants are classified into four categories i.e. Anionic, Cationic,
Nonionic and Amphoteric surfactants. Most biosurfactants are anionic or
neutral, whereas only some are cationic, such as those that contain amine
groups. The hydrophobic moiety is characterised by long- chain fatty acids
and the hydrophilic moiety may be a carbohydrate, amino acid, cyclic peptide,
phosphate, carboxyl acid or alcohol (Bognolo, 1999).

Biosurfactants are commonly classified based on their biochemical nature or

the microbial producer species. With regard to structure, these compounds
are classified into five main groups (Rahman and Gakpe, 2008).
• Glycolipids – the degree of polarity depends on the hydrocarbons used
as substrate; examples: rhamnolipids produced by Pseudomonas aeruginosa
and sophorolipids produced by species of Candida.

• Lipopolysaccharides – which normally have a high molecular mass and

are soluble in water; example: emulsan, an extracellular emulsifier produced
from hydrocarbons by the bacteria Acinotobacter calcoaceticus;

• Lipopeptides – example: surfactin produced by Bacillus subtilis

(one of the most potent biosurfactants reported in the literature);

• Phospholipids – structures common to many microorganisms; example:

biosurfactant from Corynebacterium lepus;

• Fatty acids, neutral lipids (some classified as glycolipids) and

hydrophobic proteins.

Some Microbial Amphiphiles and there producers given below

Biosurfactants microorganisms Reference

Glycolipids

Rhamnolipids Pseudomonas aeruginosa Hisatsuka et al. (1971)

P.chlororaphis Gunther et al. (2005)

Aceine tobacter Rooney et al. (2009)

calcoaceticus
Trehalose lipids R.erythropolis Rapp et al. (1979)

Arthrobacter sp. Li et al. (1984)

Mycrobacterium sp. Cooper et al. (1989)

Biosurfactant Microorganism Reference

Sophrolipids C.bombicola Cavalero and Cooper

(2003)
C.borgoriensis Cutler and Light
(1979)
T.apicola Hommel et al. (1987)
Ustilago maydis Hewald et al. (2005)
Mannosylerythritol C.antarctica Kitamoto et al. (1992)
lipids
Cellobio lipids T. petrophylum Cooper and Paddock
(1983)
U.zeae Boothroyd et al.
(1956)
Lipopeptides

Surfactin B. subtilis Arima et al. (1968)

Viscosin P. fluorescens Neu et al. (1990)

Lichenysin B. licheniformis Yakimov et al. (1995)

Serrawettin S. marcescens Matsuyama et al.

(1992)
subtilisin B. subtilis Bernheimer and
Avigad (1970)
Arthrofactin Arthrobacter sp. Strain MIS38 Morikawa et al. (1993)

Plipastatin B. subtilis Volpon et al. (2000)

Amphisin Pseudomonas sp. Sorensen et al. (2001)

Putisolvin P. putida Kuiper et al. (2004)

Surface active antibiotics

Gramicidin B. brevis Marahiel et al. (1977)

Polymyxin B. polymyxa Suzuki et al. (1965)

Fatty acids C. lepus Cooper et al. (1978)

Neutral lipids N. erythropolis MacDonald et al.

(1981)
Flavolipids Flavobacterium sp. Strain Bodour et al. (2004)
MTN11
Phospholipids T. thiooxidans Beebe and Umbreit
(1971)
Polymeric surfactants

Emulsan A.calcoaceticus Zosim et al. (1982)

P. fluorescens Persson et al. (1988)

Alasan A. radioresistens Navon-Venezia et al.

(1995)
Liposan C. lipolytica Cirigliano and Carman
(1985)
Lipomanan C. tropicalis Kappeli et al. (1984)

Biodispersan A. calcoaceticus Rosenberg et al.

(1988)
Yansan Yarrowia lipolytica Amaral et al. (2006)
Particulate biosurfactants
Vesicles A. calcoaceticus Kappeli and Finnerty
(1979)
Emulcyan Phormidium J-1 Fattom and Shilo
(1985)
GLYCOLIPIDS-

The most commonly described microbial surfactants are the glycolipids.

Glycolipids are carbohydrates (hydrophilic moiety) linked with long-chain
aliphatic acids or hydroxy aliphatic acids (hydrophobic moiety). The linkage is
by means of either ether or an ester group. The best studied glycolipids are
rhamnolipids, sophorolipids, and trehalolipids (Desai and Banat, 1997).

RAHMNOLIPIDS-

Rhamnolipids are extracellular biosurfactants mainly produced by

Pseudomonas aeruginosa. It can produce up to 100 g/L of rhamnolipids, and
hence, its cost becomes competitive against the production cost of synthetic
surfactants (Maier and Soberón-Chávez, 2000).
 Major forms of rhamnolipid contain one (monorhamnolipid) or two
(dirhamnolipid) rhamnose sugars linked to one or two 3-hydroxydecanoic acid
moieties. At first, Jarvis and Johnson reported the production of rhamnose-
containing glycolipids by P. aeruginosa in peptone-glycerol broth. The
glycolipid was found to contain two units each of l-rhamnose and
hydroxydecanoic acid (Jarvis and Johnson,1949).

Mostly, P. aeruginosa strains produce rhamnolipid mixtures, which are influ-

enced by the carbon source used, and they vary in the length and the
saturation of the fatty acid moiety (Herman and Maier, 2002). Benincasa et al.
reported that up to 28 different structural homologues of rhamnolipids were
produced by P. aeruginosa strains (Benincasa et al., 2004). Difference in the
form and the composition of the rhamnolipids produced causes variation in
physicochemical properties. Rhamnolipids are low-molecular weight
biosurfactants, which are capable of reducing ST and inter- facial tension (IFT).
When excreted into the medium, they can emulsify hydrocarbon or other
hydrophobic substrates and thereby make them available for cell metabolism.
Rhamnolipids are also reported to have antimicrobial activity against several
bacterial, yeast, and fungal strains (Benincasa et al., 2004).

SOPHOROLIPIDS-

Sophorolipids are largely produced by yeasts, mainly Candida sp. The strain
Candida bombicola ATCC 22214 can produce over 400 g/L sophorolipids and
now is used for commercial production and applications (Pekin et al., 2005).
Sophorolipids are composed of a disaccharide sophorose, -glycosidically
linked to a long chain hydroxy fatty acid. Sophorolipid can occur in the acidic
form, in which the fatty acid tail is free, or in the lactonic form, where the
carboxylic end of the fatty acid is connected to the sophorose head by
internal esterification. Sophorolipid mixtures also vary in fatty acid chain
length, and the variation depends on the type of hydrophobic organic
substrate utilized by the microorganism for growth. Properties of sophorolipid
mix- tures depend on their form and composition. For instance, lactonic
sophorolipids have better ST lowering and antimicrobial activity, whereas the
acidic ones display a better foam production and solubility (Van Bogaert et al.,
2011).
TREHALOLIPIDS-

Different types of trehalolipids are known to be produced by several

microorganisms, including the genera of Mycobacterium, Rhodococcus,
Arthrobacter, Nocardia, and Gordonia. Among the trehalolipids, the trehalose
esters produced by Rhodococcus erythropolis have been studied most
extensively. Trehalolipids consist of a disaccha- ride trehalose linked to
fattyacid groups by ester bond. The ester of fatty acids with trehalose was
first noticed by Anderson and Newman in 1933, but not as a surface active
agent (Anderson and Newman, 1933). Trehalolipids caught the attention as a
general surfactant after an emulsion layer containing trehalose dimycolates
was discovered in Arthrobacter paraffineus culture broths when the cells
were grown on hydrocarbon substrates (Shao, 2011). Trehalolipids are most
widely used in bioreme- diation technologies as such compounds are known
to enhance the bioavailability of hydrocarbons (Franzetti et al., 2010).

LIPOPEPTIDES-

Lipopeptides consist of short linear chains or cyclic structures of amino acids,

linked to a fatty acid via ester or amide bonds or both. Lipopeptides contain
the rare and modified amino acids, which are not used for ribosomal protein
synthesis (Sen, 2010). They vary in terms of the types of amino acids present
in the peptide ring, as well as in the chain length and structure of the fatty
acid component.

SURFACTIN-

Surfactin, the most studied and potent lipopeptide biosurfactant, is produced

by various strains of Bacillus subtilis. The cyclic lipopeptide surfactin consists
of 3-hydroxy- 13-methyl-tetradecanoic acid amidated to the N-terminal amine
of a heptapeptide moiety with the carboxy terminal end of the peptide being
further esterified to the hydroxyl group of the fatty acid (Sen, 2010). Surfactin
was discovered by Arima et al. from the culture broth of B. subtilis as a potent
inhibitor of blood clotting, but was also found to be a powerful surface active
agent (Arima et al., 1968). Besides surfactant property, surfactin possesses a
number of biological activities, which makes it a versatile biomolecule with
tremendous commercial application potentials.

LICHENYSIN-

Lichenysins are most potent lipopeptide biosurfactants produced by B.

licheniformis and are named lichenysin A, B, C, D, G, and surfactant BL86 with
regard to pro- ducing strains. The production of lichenysin was first described
by Jenneman et al. in a halotolerant Bacillus sp., which was then identified as
B. licheniformis JF-2 (Jenneman et al., 1983). Lichenysin A was reported to
exhibit critical micelle concentration (CMC) of 12 mg/L, which is around one-
half of the CMC of sur factin (25 mg/L). Rationale behind this fact was the less
polar peptide moiety and the presence of a longer hydroxy fatty acid giving
lichenysin A delicate hydrophile-lipophile balance (Yakimov et al., 1995).