Banat 2010

Appl Microbiol Biotechnol (2010) 87:427–444
DOI 10.1007/s00253-010-2589-0
MINI-REVIEW
Microbial biosurfactants production, applications

and future potential
Ibrahim M. Banat & Andrea Franzetti & Isabella Gandolfi & Giuseppina Bestetti &
Maria G. Martinotti & Letizia Fracchia & Thomas J. Smyth & Roger Marchant
Received: 14 February 2010 / Revised: 24 March 2010 / Accepted: 24 March 2010 / Published online: 28 April 2010
# Springer-Verlag 2010
Abstract Microorganisms synthesise a wide range of Keywords Biosurfactants . Bioemulsifiers . Surfactants .

surface-active compounds (SAC), generally called biosurfac- Emulsifiers
tants. These compounds are mainly classified according to
their molecular weight, physico-chemical properties and
mode of action. The low-molecular-weight SACs or biosur- Introduction
factants reduce the surface tension at the air/water interfaces
and the interfacial tension at oil/water interfaces, whereas the Microbial surface-active compounds
high-molecular-weight SACs, also called bioemulsifiers, are
more effective in stabilising oil-in-water emulsions. Biosur- Microbial surface-active compounds are a group of structur-
factants are attracting much interest due to their potential ally diverse molecules produced by different microorganisms
advantages over their synthetic counterparts in many fields and are mainly classified by their chemical structure and
spanning environmental, food, biomedical, and other indus- their microbial origin. They are made up of a hydrophilic
trial applications. Their large-scale application and produc- moiety, comprising an acid, peptide cations, or anions,
tion, however, are currently limited by the high cost of mono-, di- or polysaccharides and a hydrophobic moiety of
production and by limited understanding of their interactions unsaturated or saturated hydrocarbon chains or fatty acids.
with cells and with the abiotic environment. In this paper, we These structures confer a wide range of properties, including
review the current knowledge and the latest advances in the ability to lower surface and interfacial tension of liquids
biosurfactant applications and the biotechnological strategies and to form micelles and microemulsions between two
being developed for improving production processes and different phases. These compounds can be roughly divided
future potential. into two main classes (Neu 1996): low-molecular-weight
compounds called biosurfactants, such as lipopeptides,
glycolipids, proteins and high-molecular-weight polymers of
I. M. Banat (*) : T. J. Smyth : R. Marchant polysaccharides, lipopolysaccharides proteins or lipoproteins
School of Biomedical Sciences, University of Ulster,
that are collectively called bioemulsans (Rosenberg and Ron
Coleraine BT52 1SA, Northern Ireland, UK
e-mail: [email protected] 1997) or bioemulsifiers (Smyth et al. 2010b). The former
group includes molecules which can efficiently reduce
A. Franzetti : I. Gandolfi : G. Bestetti surface and interfacial tension, while the latter are amphiphil-
Department of Environmental Sciences,
ic and polyphilic polymers which are usually more effective
University of Milano-Bicocca,
Piazza della Scienza 1, in stabilising emulsions of oil-in-water but do not lower the
20126 Milano, Italy surface tension as much (Smyth et al. 2010a).
The best-studied microbial surfactants are glycolipids.
M. G. Martinotti : L. Fracchia
Among these, the best-known compounds are rhamnoli-
DiSCAFF, Università del Piemonte Orientale,
Via Bovio, 6, pids, trehalolipids, sophorolipids and mannosylerythritol
28100 Novara, Italy lipids (MELs) (Fig. 1), which contain mono- or disac-
428 Appl Microbiol Biotechnol (2010) 87:427–444
Monorhamnolipids Dirhamnolipid
Acidic Sophorolipid Lactonic Sophorolipid
Trehalose monomycolates
Trehalose dimycolates
Surfactin
Mannosylerythritol lipids
CH3
(CH2)8
CHOH
CH3
CH2
(CH2)9
C O
CHOH O
O
C O O O
C CH2
O
HO O O
CH2 O O
O
O
NH HO NH
C O C O
HO NH n
(CH ) CH3
2 12
C O
CH3
CH3
Emulsan
Fig. 1 Chemical structure of the most studied microbial surface-active compounds; mono- and dirhamnolipids, acidic and lactonic sophorolipids,
monomycolates trehalose lipid and dimycolates trehalose lipids, mannosylerythritol lipids, surfactin and finally emulsan
Appl Microbiol Biotechnol (2010) 87:427–444 429
charides, combined with long-chain aliphatic acids or Biosurfactant-enhanced bioremediation

hydroxyaliphatic acids. Rhamnolipid production by Pseu-
domonas species has been extensively studied, and Amphiphiles are able to alter the physico-chemical con-
potential applications have been proposed (Maier and ditions at the interfaces affecting the distribution of the
Soberón-Chávez 2000). Rhamnolipids from Pseudomonas chemicals among the phases (Tiehm 1994). For instance, a
aeruginosa are currently commercialised by Jeneil Bio- hydrocarbon-contaminated soil contains at least six phases:
surfactant, USA, mainly as a fungicide for agricultural bacteria, soil particles, water, air, immiscible liquid and
purposes or an additive to enhance bioremediation solid hydrocarbon. The hydrocarbons can be partitioned
activities. Trehalolipids are produced by a number of among different states: solubilised in the water phase, ad/
different microorganisms, such as Mycobacterium, Nocar- absorbed to soil particle, sorbed to cell surfaces and as a
dia and Corynebacterium. However, the most extensively free/insoluble phase. Biosurfactants added to this system
studied compounds in this class are trehalose dimycolates can interact with both the abiotic particles and the bacterial
produced by Rhodococcus erythropolis (Rapp et al. 1979). cells.
Sophorolipids, on the other hand, are produced mainly by This affects the mechanisms of interaction with environ-
yeasts, such as Candida bombicola (also known as ments with regard to the micellarisation and emulsification
Torulopsis bombicola), Centrolene petrophilum, Candida of organic contaminants, the interaction with sorbed
apicola and Rhodotorula bogoriensis, while MELs are contaminants and the sorption to soil particles which leads
produced by Pseudozyma yeasts, Pseudozyma aphidis, to the alteration of cell-envelope composition and hydro-
Pseudozyma antarctica and Pseudozyma rugulosa phobicity. The interactions between micelles and cells are
(Konishi et al. 2007a, b). Cyclic lipopeptides are produced among the main alterations to the bacterial component
by a number of Bacillus species as antibiotic molecules. (Volkering et al. 1998). These phenomena on the one hand
Among these, the most important compound is surfactin can be exploited to increase the bioavailability of poorly
produced by Bacillus subtilis because of its very high soluble contaminants, thus increasing biodegradation rate,
activity (Desai and Banat 1997; Rosenberg and Ron 1999). or on the other hand, can result in an inhibition of
A wide variety of microorganisms, including some Archaea, biodegradation.
produce high-molecular-weight polymers, the most exten- In spite of the publication bias which favours an over-
sively investigated being bioemulsans (Fig. 1) which are publication of successful applications, the main emerging
synthesised by various species of Acinetobacter. The first feature of the large body of literature in this area is the
studied compound was RAG-1 emulsan, an amphiphilic contrasting result reported on efficiency. For instance,
polysaccharide produced by Acinetobacter calcoaceticus rhamnolipids can stimulate the degradation of n-hexade-
RAG-1, which is also the only commercially available cane by the producer strain P. aeruginosa, but didn’t
bioemulsifier at present (Suthar et al. 2008). stimulate degradation by Rhodococcus strains showing
strain specificity. In contrast, biosurfactants from R.
erythropolis strain 3C-9 significantly increased the degra-
Potential applications dation rate of n-hexadecane by two phylogenetically distant
species, Alcanivorax dieselolei and Psychrobacter celer, in
Environmental applications flask tests (Noordman and Janssen 2002; Peng et al. 2007).
Therefore, with the current state of knowledge, the modelling
In many cases, environmental contamination caused by of the effect of biosurfactant addition in bioremediation
industrial activity is due to accidental or deliberate release treatment is not predictable, and efficacy has to be evaluated
of organic and/or inorganic compounds into the environ- experimentally (Franzetti et al. 2006, 2008b). To gain better
ment. Such compounds pose problems for remediation, as insight into this problem, it is useful to review the current
they become easily bound to soil particles. The application knowledge and recent advances regarding these interactions.
of biosurfactants in the remediation of organic compounds, For excellent reviews about interactions between surfac-
such as hydrocarbons, aims at increasing their bioavailabil- tants and the environment, see Volkering et al. (1998) and
ity (biosurfactant-enhanced bioremediation) or mobilising Paria (2008). The interactions between bacteria, contami-
and removing the contaminants by pseudosolubilisation and nants and biosurfactant can be interpreted from a functional
emulsification in a washing treatment. The application of perspective, considering that the main natural role attributed
biosurfactants in the remediation of inorganic compounds to biosurfactants is their involvement in hydrocarbon
such as heavy metals, on the other hand, is targeted at uptake (Perfumo et al. 2010a). Microbial surfactants can
chelating and removal of such ions during a washing step promote the growth of bacteria on hydrocarbons by
facilitated by the chemical interactions between the amphi- increasing the surface area between oil and water and
philes and the metal ions. through emulsification and increasing pseudosolubility of
hydrocarbons through partitioning into micelles (Miller and with biodegradation similarly inhibited by Tween 20,
Zhang 1997; Volkering et al. 1998). sodium dodecyl sulfonate, tetradecyl trimethyl ammonium
High-molecular-weight biosurfactants (bioemulsifiers) bromide and Citrikleen at concentrations equal or greater
have great potential for stabilising emulsions between than their CMCs (Billingsley et al. 1999).
liquid hydrocarbons and water, thus increasing the surface Another proposed role of biosurfactants in hydrocarbon
area available for bacterial biodegradation. However, they uptake is the regulation of cell attachment to hydrophobic
have been rarely tested as enhancers of hydrocarbon and hydrophilic surfaces by exposing different parts of cell-
biodegradation in bioremediation systems, and contrasting bound biosurfactants, thus changing cell-surface hydropho-
results are reported in the literature (Barkay et al. 1999; bicity (Rosenberg et al. 1987; Franzetti et al. 2008a).
Franzetti et al. 2009a). This natural role can be exploited by adding (bio)
For low-molecular-weight biosurfactants, above the Crit- surfactants to increase the hydrophobicity of degrading
ical Micelle Concentration (CMC), a significant fraction of microorganisms and to allow cells’ easier access to
the hydrophobic contaminant partitions in the surfactant hydrophobic substrates (Shreve et al. 1995). The release
micelle cores. In some cases, this results in a general increase of LPS by Pseudomonas spp. induced by sub-CMC levels
in the bioavailability of contaminants for degrading micro- of rhamnolipids allowed a more efficient uptake of
organisms. Successful applications of rhamnolipids and hexadecane by rendering the cell surface more hydrophobic
surfactin in enhanced bioremediation have been recently (Al-Tahhan et al. 2000). Noordman and Janssen (2002)
reviewed (Mulligan 2009). In addition, Wang and Mulligan reported that rhamnolipid produced by P. aeruginosa UG2
(2009) studied the effect of ammonium ion concentration and facilitated the hydrocarbon uptake of the producer strain
pH on the potential application of rhamnolipid and surfactin and increased the degradation of hexadecane, while the
for enhanced biodegradation of diesel. A lipopeptide and same product did not stimulate to the same extent the
protein–starch–lipid produced by two strains of P. aeruginosa biodegradation of hexadecane by four unrelated species
significantly enhanced the solubilisation of phenanthrene, (Acinetobacter lwoffii RAG1, R. erythropolis ATCC 19558,
pyrene and fluorene, increasing their metabolism and R. erythropolis DSM 43066 and strain BCG112), nor was
supporting sustained growth (Bordoloi and Konwar 2009). degradation of hexadecane stimulated by addition of the
Polycyclic Aromatic Hydrocarbons (PAH) biodegradation biosurfactants produced by these species themselves.
was also investigated by Das et al. (2008b); they used Bacillus Zhong et al. (2007) showed that the adsorption of
circulans to increase the bioavailability of anthracene. dirhamnolipid biosurfactants on cells of B. subtilis, P.
Interestingly, the organism had better growth and biosurfac- aeruginosa and Candida lipolytica depended on the
tant production on glycerol containing mineral medium physiological status of the cells and was specific to the
supplemented with anthracene, although it was unable to microorganisms. Furthermore, the biosurfactant adsorp-
utilise anthracene as the sole carbon source. These authors tion affected the cell-surface hydrophobicity depending
were able to demonstrate, however, that anthracene was used on the rhamnolipid concentration and the physiological
as a substrate for the production of the biosurfactant. state of the cell. The effect of exogenous rhamnolipids
The specific modes of hydrocarbon uptake, however, are on cell-surface composition of P. aeruginosa NBIMCC
not fully understood. Recently Cameotra and Singh (2009) 1390 was recently studied by Sotirova et al. 2008. They
elucidated the mechanism of n-hexadecane uptake mediated showed that above the CMC, rhamnolipids caused a 22%
by rhamnolipids in P. aeruginosa. The rhamnolipids reduction of total cellular LPS content, while at concen-
produced an emulsion with hexadecane, thus facilitating trations below the CMC, they caused changes in the
increased contact between the hydrocarbon substrate and bacterial outer membrane protein composition yet did not
the bacteria. It was also observed that uptake of the affect the LPS component.
biosurfactant-coated hydrocarbon droplets occurred, sug- Chang et al. (2009) demonstrated that the cell-surface
gesting a mechanism like pinocytosis taking place, a hydrophobicity was enhanced by the accumulation at the
process not previously reported in bacterial hydrocarbon cell surface of different fatty acids during growth on
uptake systems. hydrocarbon in R. erythropolis NTU-1. A significant
In contrast, it is well known that the presence of a correlation between the modification of the cell surface by
surfactant can detrimentally affect biodegradation. Micelle saponins and the degree of hydrocarbon biodegradation was
cores can trap organic contaminants, creating a barrier reported by Kaczorek et al. (2008).
between microorganisms and organic molecules, resulting
in the potential substrate becoming less rather than more Biosurfactant-enhanced soil washing
available. For example, Witconol SN70, a non-ionic
alcohol ethoxylate surfactant (Colores et al. 2000), reduced The application of microbial SACs to remove contami-
the biodegradation rate of hexadecane and phenanthrene, nants from soils is a technology characterised by less
uncertainty than biosurfactant-enhanced bioremediation, stimulate biosurfactant production in situ by indigenous

since the removal efficiency is mainly driven by the bacteria (Singh et al. 2007).
chemico-physical properties of the biosurfactants and not At present, the first strategy is the most exploited. In
their effect on metabolic activity or changes in cell-surface practice, a major obstacle to the in situ production of
properties. However, the mechanisms affecting hydrocar- biosurfactants is the difficulty of isolating microbial strains
bon mobilisation or removal from soils resemble those adapted to the extreme environment of the reservoirs, which
involved in enhancing bioavailability for bioremediation. features high pressure, salinity, temperatures up to 85°C
The ability to stabilise oil/water emulsions and increase and extreme pH values. All the main types of microbial
hydrocarbon solubility enhances biodegradation and hy- surface-active compounds have been proposed for a MEOR
drocarbon removal from soils (Franzetti et al. 2010). These application. Although rhamnolipids have been most fre-
mobilisation and solubilisation effects occur at concen- quently used, lipopeptides, such as surfactin, lichenysin and
trations above and below the CMC. Mulligan (2009) emulsan have also proved very effective in enhancing oil
reviewed the application of biosurfactants in enhanced soil recovery (Sen 2008). Xu et al. (2009) further demonstrated
washing for hydrocarbon- and metal-contaminated soils. the effectiveness of a polysaccharide produced by Strepto-
More recently, Franzetti et al. (2009b) reported efficient coccus sp. BT-003 for the same application. The use of
removal of crude oil from soil using extracellular bio- biosurfactants in MEOR has been extensively reviewed
emulsifier produced by Gordonia sp. BS29. Interesting (Banat et al. 2000; Singh et al. 2007; Sen 2008). Recently,
papers reporting enhanced metal removal or mobilisation several laboratory experiments were carried out to evaluate
have been published in the past 2 years. Biosurfactants are the effectiveness of biosurfactant-assisted oil extraction and
efficient in removing bulk arsenic from mine tailings or recovery. Biosurfactants produced by R. erythropolis and
contaminated soils under alkaline conditions (Wang and Rhodococcus ruber were used to extract hydrocarbons from
Mulligan 2009). Das et al. (2009) showed that cadmium oil shale in flask experiments; the maximum recovery was
removal from aqueous solution also occurred at concen- 25% and 26% for the two strains, respectively, with even
trations less than the CMC, while at a concentration of five lower recovery when a high percentage of asphaltenes and
times, the CMC resulted in almost complete removal of resin compounds were present in the oil (Haddadin et al.
100 ppm of metal ions. Wen et al. (2009) studied 2009).
rhamnolipid degradation in soils contaminated by Cd and When the main purpose of the laboratory evaluation is a
Zn, they suggested that rhamnolipid in the soil remain long screening to test candidate molecules for full-scale applica-
enough to enhance metal phytoextraction, yet not long tion, most experiments are currently performed in sand-
enough to raise concerns regarding metal transport in the packed column systems to simulate oil reservoir conditions.
long term. Pornsunthorntawee et al. (2008) demonstrated that both B.
subtilis PT2 and P. aeruginosa SP4 biosurfactants were
Industrial applications more effective than three synthetic surfactants in oil
recovery from a sand-packed column, the B. subtilis
The main industrial application for biosurfactants is in the product being the most effective with an oil removal of
field of oil recovery and processing. Since traditional oil 61% against 57% for the P. aeruginosa surfactants and
recovery technologies can only recover approximately 40– about 4% when distilled water was used. Some other
45% of the oil present in the reservoir, some technologies, microbial surface-active compounds were also tested in
collectively defined as enhanced oil recovery (EOR), have column systems. For example, a bioemulsifier from
been developed (Banat 1995; Dastgheib et al. 2008). Bacillus licheniformis K125, which reduced the surface
Among these, microbial EOR (MEOR), which takes tension to 34 mN/m, gave about 43% additional oil
advantage of microbial production of surface-active com- recovery after water extraction (Suthar et al. 2008), while
pounds, is considered to have the most cost-effective some biosurfactants from different strains of P. aeruginosa
potential (Sen 2008). When poor oil recovery from oil gave 49–62% oil recovery (Bordoloi and Konwar 2008).
wells is due to either low permeability of the rocks forming Although offsite biosurfactant production is the most
the reservoir or to the high viscosity of the crude oil, the common practice in MEOR, its potential has not been fully
ability of biosurfactants to reduce the oil/water interfacial realised yet due to its high cost. To reduce costs, Wang et
tension and to form stable emulsions can improve the al. (2007) suggested selecting Pseudomonas strains, which
process efficiency. Three different strategies have been can efficiently grow on renewable low-cost substrates and
identified for MEOR: biosurfactant production in offsite genetically engineer them to produce high yields of
reactors and subsequent addition to the oil reservoir; rhamnolipid. The prospect for such a strategy is probably
biosurfactant production by injected allochthonous micro- quite poor since the production of rhamnolipids in
organisms; injection of nutrients into the reservoir to Pseudomonas regulated through the quorum sensing
system and genetic intervention is difficult. However, an are also used to retard staling, solubilise flavour oils and
alternative would be an in situ production of biosurfac- improve organoleptic properties in bakery and ice cream
tants, either by injected bacteria or by stimulated autoch- formulations and as fat stabilisers during cooking of fats.
thonous microorganisms. Therefore some effort has Although the addition of rhamnolipids has been suggested
recently been put into isolating new surfactant-producing to improve dough characteristics of bakery products, the
microbial strains using extreme conditions to reproduce use as food ingredients of compounds derived from an
those encountered in oil reservoirs (Agarwal and Sharma opportunistic pathogen such as P. aeruginosa is not
2009), while other research has focused on selective practically feasible. Instead, it has been suggested to use
activation of indigenous microorganisms able to enhance biosurfactants obtained from yeasts or Lactobacilli, which
oil recovery (Bao et al. 2009). The potential utilisation of are generally recognised as safe and are already involved
selected exogenous microoganisms can be assessed either in several food-processing technologies (Nitschke and
in the laboratory or directly in the field. The performance Costa 2007).
of two bacteria, B. subtilis and Leuconostoc mesenteroides, Wetting, dispersing and surface-tension reduction prop-
biosurfactant- and exopolymer-producing strains, respec- erties, as well as low toxicity and high biodegradability,
tively, were evaluated by using oil-saturated glass micro- suggested the application of biosurfactants, especially
models of a fractured porous medium to determine oil glycolipids, as components of detergent formulations.
recovery. B. subtilis gave better oil recovery due to the Low-foaming sophorolipids from C. bombicola appear
reduction of oil viscosity and the interfacial tension suitable due to their high detergency ability, low cytotox-
(Soudmand-Asli et al. 2007). icity and high biodegradability and general environmentally
While a number of field trials of in situ applications of acceptable properties (Hirata et al. 2009). Also, cyclic
MEOR are reported in the literature (see Sen 2008 for a lipopeptide biosurfactants from B. subtilis improved wash
review), it has not been completely elucidated whether performance by acting additively with other detergent
introduced microorganisms can actually be effective in oil components. Since they have shown better results at low
recovery or if they are out-competed by indigenous temperature, such formulations are promising from an
bacteria. The inability to compare test wells with control energy-saving point of view, allowing laundering at lower
wells subjected to similar treatment procedures without temperatures (Mukherjee 2007). Several surfactant–enzyme
introducing live microorganisms or products makes valid mixtures were tested for rubisco removal from both
conclusions difficult to draw. To provide better insight into hydrophobic and hydrophilic surfaces; the most effective
the dynamics of the microbial community, Wang et al. formulation was a surfactin–subtilisin A detergent, thus
(2008) monitored changes in the community using demonstrating that it would be possible to generate fully
molecular markers by denaturing gradient gel electropho- renewable cleaning formulations with good performance
resis in an oil reservoir during a process of MEOR. They (Onaizi et al. 2009).
observed that both exogenous and stimulated indigenous
bacteria appeared to contribute to the increased oil Biomedical applications
recovery.
Beside applications in MEOR, microbial surface-active Biosurfactants, when present in heterogeneous systems, tend
compounds can also be exploited for other applications in to aggregate at the phase boundaries or interfaces. Organic
the oil industry. For example, the de-emulsifying proper- molecules present in aqueous phase are known to be inclined
ties shown by some biosurfactant-producing microorgan- to immobilise at the solid interface is such interfacial
isms may be used to break emulsions which form at systems. They eventually form a conditioning film, which
various steps in oil extraction and processing, thus will affect the properties (surface energy and wettability) of
allowing a better recovery of the product. The surface- the original surface (Neu 1996). In a similar manner to
tension decrease produced by microbial surfactants can organic-conditioning films, biosurfactants partition at the
also be used to separate oil from tank bottom sludge interfaces and can affect the adhesion properties of micro-
(Singh et al. 2007; Joseph and Joseph 2009; Perfumo et al. organisms. Another function valuable for medical application
2010b). is their ability to disrupt membranes leading to cell lysis
Due to their physico-chemical properties, the use of through increased membrane permeability leading to metab-
microbial surface-active compounds has also been pro- olite leakage. This occurs due to changes in physical
posed for various industrial applications, as additives in membrane structure or through disrupting protein conforma-
foods, cosmetics and detergent formulations (Banat et al. tions which alters important membrane functions such as
2000). In the food industry, the most useful property is the transport and energy generation (Van Hamme et al. 2006;
ability to form stable emulsions, which improves the Ortiz et al. 2008, 2009; Sotirova et al. 2008; Sánchez et al.
texture and creaminess of dairy products. Biosurfactants 2009, 2010; Zaragoza et al. 2009).
Antimicrobial activity of biosurfactants against methicillin and vancomycin) in tested strains. In

addition, flocculosin was able to kill C. albicans cells in a
The high demand for new antimicrobial agents following very short period of time. Huang et al. (2007) observed
increased resistance shown by pathogenic microorganisms that a lipopeptide antimicrobial substance produced by the
against existing antimicrobial drugs has drawn attention to strain B. subtilis fmbj, which is mainly composed of
biosurfactants as antibacterial agents (Bĕhal 2006). Some surfactin and fengycin, was able to inactivate endospores
biosurfactants have been reported to be suitable alternatives of B. cereus. Observation by TEM indicated that the
to synthetic medicines and antimicrobial agents and may lipopeptide could damage the surface structure of the
therefore be used as effective and safe therapeutic agents spores.
(Cameotra and Makkar 2004; Singh and Cameotra 2004; Antiviral activity of biosurfactants, mainly surfactin and
Banat et al. 2000). its analogues, has also been described (Naruse et al.
Lipopeptides form the most widely reported class of 1990). The more effective inactivation of enveloped
biosurfactants with antimicrobial activity. Surfactin, pro- viruses, such as retroviruses and herpes viruses, compared
duced by B. subtilis, is the best-known lipopeptide (Arima to non-enveloped viruses, suggests that this inhibitory
et al. 1968). Other antimicrobial lipopeptides include action may be mainly due to physico-chemical interactions
fengycin, iturin, bacillomycins and mycosubtilins produced between the virus envelope and the surfactant (Vollenbroich
by B. subtilis (Vater et al. 2002). Lichenysin, pumilacidin et al. 1997). Antimicrobial lipopeptides produced by B.
and polymyxin B (Naruse et al. 1990; Yakimov et al. 1995; subtilis fmbj inactivated cell-free virus of porcine parvovi-
Grangemard et al. 2001; Landman et al. 2008) are other rus, pseudorabies virus, newcastle disease virus and bursal
antimicrobial lipopeptides produced by B. licheniformis, disease virus, while it effectively inhibited replication and
Bacillus pumilus and Bacillus polymyxa, respectively. The infectivity of the newcastle disease virus and bursal disease
production of antimicrobial lipopeptides by Bacillus pro- virus but had no effect on pseudorabies virus and porcine
biotic products is one of the main mechanisms by which parvovirus (Huang et al. 2006). Sophorolipids are also
they inhibit the growth of pathogenic microorganisms in the claimed to have activity against human immunodeficiency
gastrointestinal tract (Hong et al. 2005). Other reported virus (Shah et al. 2005). Similarly, a rhamonolipid and its
biosurfactants having antimicrobial activity are daptomycin, complex with alginate, both produced by a Pseudomonas
a cyclic lipopeptide from Streptomyces roseosporus (Baltz sp. strain, showed significant antiviral activity against
et al. 2005), viscosin, a cyclic lipopeptide from Pseudomo- herpes simplex virus types 1 and 2 (Remichkova et al.
nas (Neu et al. 1990; Saini et al. 2008), rhamnolipids 2008). The suppressive effect of the compounds on
produced by P. aeruginosa (Abalos et al. 2001; Benincasa herpes simplex virus replication was dose-dependent and
et al. 2004) and sophorolipids produced by C. bombicola occurred at concentrations lower than the critical micelle
(Kim et al. 2002; Van Bogaert et al. 2007). Mannosylery- concentration.
thritol lipids (MEL-A and MEL-B) produced by Candida The antifungal activities of biosurfactants have long
antarctica strains have also been reported to exhibit antimi- been known, although their action against human patho-
crobial action against Gram-positive bacteria (Kitamoto et al. genic fungi has been rarely described (Tanaka et al. 1997;
1993). Chung et al. 2000; Abalos et al. 2001). Recently, a
Recently, a lipopeptide biosurfactant produced by a glycolipid isolated from the yeast-like fungus P. flocculosa,
marine organism, B. circulans, was found to be active named flocculosin was shown to display in vitro antifungal
against Proteus vulgaris, Alcaligens faecalis, methicillin- activity against several pathogenic yeasts, associated with
resistant Staphylococcus aureus (MRSA) and other human mycoses (Mimee et al. 2005). This product
multidrug-resistant pathogenic strains (Das et al. 2008a) positively inhibited all pathogenic strains tested under
while not having any haemolytic activity. A rhamnolipid acidic conditions and showed synergistic activity with
surfactant produced from soybean oil waste had antimi- amphotericin B, increasing its efficacy while decreasing
crobial activity against several bacteria and fungi, namely any toxicity and other side effects.
Bacillus cereus, S. aureus, Micrococcus luteus, Mucor The antifungal activity against phytopathogenic fungi
miehei and Neurospora crassa (Nitschke et al. 2009b). has been demonstrated for glycolipids, such as cellobiose
Flocculosin, a cellobiose lipid produced by the yeast-like lipids (Teichmann et al. 2007; Kulakovskaya et al. 2009,
fungus Pseudozyma flocculosa, was tested against clinical 2010) and rhamnolipids (Debode et al. 2007; Varnier et al.
bacterial isolates and the pathogenic yeast Candida 2009), and cyclic lipopeptides (Tran et al. 2007, 2008),
albicans (Mimee et al. 2009). The glycolipid was partic- including surfactin, iturin and fengycin (Velmurugan et al.
ularly effective against Staphylococcus species, including 2009; Arguelles-Arias et al. 2009; Chen et al. 2009; Snook
MRSA, and its antibacterial activity was not influenced by et al. 2009; Mohammadipour et al. 2009; Grover et al.
the presence of common resistance mechanisms (e.g. 2010).
Biosurfactants as anti-adhesives silver used and greater antimicrobial activity. The concen-
tration of silver in the silver–biosurfactant solution was
Biofilm formation and swarming motility are the key from 129- to 258-fold less than the concentration when
microbial activities in the colonisation of a surface and silver was used alone. Based on these results, an interna-
therefore can increase the chance of nosocomial infections tional patent PCT/IB2009/055334 entitled “Biosurfactant
on different medical devices (Khardori and Yassien 1995; composition produced by a new B. licheniformis strain,
Vinh and Embil 2005; McCann et al. 2008; Harriott and uses and products thereof” has been deposited in 2009,
Noverr 2009). Current biofilm preventive strategies are inventors Martinotti M.G., Rivardo F. Allegrone G., Ceri
essentially aimed at coating medical surfaces with antimi- H., Turner R. Unpublished preliminary results obtained by
crobial agents (von Eiff et al. 2005; Basak et al. 2009). the same research group showed anti-adhesion effects of
However, recent studies have suggested that non-antibiotic two lipopeptides produced by bacterial endophytes, isolated
molecules naturally produced within bacterial communities, from oleander and rice, on the biofilm of two different
including secreted signalling molecules or surface-active pathogenic strains of C. albicans (Fig. 2).
biosurfactants, could also interfere with biofilm formation, Valle et al. (2006) observed that distinct serotypes of
modulating microbial interaction with interfaces (Neu 1996; group II capsular polysaccharides, produced by the uropa-
Federle and Bassler 2003; Rasmussen and Givskov 2006; thogenic E. coli (UPEC strain CFT073) behaved like
Rodrigues et al. 2006a). In addition to their direct action surface-active polymers that displayed anti-adhesion prop-
against pathogens, biosurfactants can also alter the physical erties. The treatment of abiotic surfaces with group II
and chemical condition of the environment where biofilms capsular polysaccharides significantly inhibit mature bio-
are developing (Mireles et al. 2001; Merk et al. 2005). film development of a broad range of Gram-positive and
Dealing with these biofilms is difficult yet an important Gram-negative bacteria.
goal, since microbes embedded within them are associated Strategies for the prevention of microbial biofilm formation
with many infections and usually become difficult to treat on silicone rubber voice prostheses on vinyl urethral catheters
effectively with traditional antimicrobials (Morikawa and on other material have also been described (Velraeds et al.
2006). 2000; Mireles et al. 2001; Rodrigues et al. 2004, 2006b, c,
Recently, the capability of two lipopeptide biosurfac- 2007).
tants, produced by B. subtilis V9T14 and B. licheniformis Beside treatment of medical devices, biosurfactants have
V19T21, to inhibit biofilm adhesion of pathogenic bacteria been used in the pre-treatment of material surfaces found in
to polystyrene was demonstrated using the MBEC device food-processing environments. Listeria monocytogenes,
(Rivardo et al. 2009). The two biosurfactants V9T14 and Salmonella enteritidis and Enterobacter sakazakii are
V19T21 showed interesting specific anti-adhesion activity examples of pathogenic bacteria implicated in outbreaks
being able to selectively inhibit biofilm formation by two associated with the ingestion of contaminated food.
pathogenic strains. In particular, S. aureus ATCC 29213 Numerous studies have shown that these bacteria are able
and Escherichia coli CFT073 biofilm formation were to adhere and to form biofilms on food-contact surfaces that
decreased by 97% and 90%, respectively. V9T14 biosur- are more resistant to sanitation than free-living cells
factant active on the Gram-negative strain was ineffective (Kalmokoff et al. 2001; Stepanovic et al. 2004; Kim et al.
against the Gram-positive and the opposite for V19T21 2006). The pre-conditioning of surfaces using microbial
biosurfactant. This activity was observed either by coating surface-active compounds could be an interesting strategy
the polystyrene surface or by adding the biosurfactant to the to prevent adhesion of food-borne pathogens to solid
inoculum. surfaces.
Two fractions from each purified biosurfactant, obtained Meylheuc et al. (2006b) demonstrated that the pre-
by flash chromatography, fractions (I) (surfactin) and (II) conditioning of stainless steel and polytetrafluoroethylene
(fengycin), showed that fraction (II) was responsible for the surfaces with an anionic biosurfactant produced by Pseu-
anti-adhesion activity in both strains. Moreover the V9T14 domonas fluorescens reduced the number of L. monocyto-
biosurfactant has been shown to increase biofilm eradica- genes LO28-adhering cells and thus resembled the
tion efficacy of different antibiotics against a urinary tract- bactericidal activities of the disinfectants sodium hypochlo-
infective E. coli strain (Martinotti et al. 2009—deposited rite (NaOCl) and peracetic acid/hydrogen peroxide (PAH).
patent). More recently, the activity of AgNO3 combined Similarly, the ability of adsorbed biosurfactants obtained
with the lipopeptide biosurfactant V9T14 has been studied from Lactobacillus helveticus and P. fluorescens to inhibit
against a preformed E. coli biofilm on the Calgary Biofilm the adhesion of four Listeria strains to stainless steel was
device (Rivardo et al. 2010). Results indicated that the investigated (Meylheuc et al. 2006a). Whichever strain of
activity of silver can be synergistically enhanced by the L. monocytogenes used in combination with biosurfactants,
presence of V9T14, allowing a reduction in the quantity of the anti-adhesive biological coating developed both reduced
Fig. 2 Anti-adhesion activity of

biosurfactants OC5 and RA1
produced by two bacterial
endophytes against biofilm pro-
duced by C. albicans strains.
Shaded columns, untreated con-
trols; white columns, treatments
with different biosurfactants
concentrations. Vertical bars
show the standard deviation of
the mean based on three inde-
pendent measurements. *P<
0.05 calculated by t test
the total adhering flora and the viable and culturable aureus biofilm integrity and formation. Moreover, surfac-
adherent bacteria on stainless steel surfaces. More recently, tant addition to preformed mature biofilms accelerated their
another group investigated the effect of rhamnolipids and dispersal and altered the characteristics of the biofilm
surfactin biosurfactants on the adhesion of the food morphology.
pathogens E. sakazakii, L. monocytogenes and S. enteritidis Another interesting application field for probiotics that is
to polypropylene and stainless steel surfaces (Nitschke et al. gaining more interest is their use in preventing oral
2009a). Preconditioning with surfactin, rather than rhamno- infections. The role of probiotics on oral health has been
lipid, caused a reduction in the number of adhering cells thoroughly investigated (Çaglar et al. 2005; Meurman
particularly of L. monocytogenes and to some extent E. 2005; Meurman and Stamatova 2007; Hatakka et al.
sakazakii on stainless steel. Surfactin showed a significant 2007; Kõll et al. 2008). Van Hoogmoed et al. (2004)
decrease in the adhesion on polypropylene of all strains. demonstrated that Streptococcus mitis biosurfactant
The adsorption of surfactin on polystyrene also reduced the inhibited adhesion of Streptococcus sobrinus HG 1025
adhesion of S. enteritidis- and L. monocytogenes-growing and Streptococcus mutans ATCC 25175 to bare enamel,
cells. In addition, surfactin was able to delay bacterial while S. mitis biosurfactant was able to inhibit the adhesion
adhesion within short contact periods using non-growing of S. sobrinus HG 1025 to salivary pellicles. The authors
cells or longer contact periods using growing cells. later reported that these reductions may be attributed to
Probiotics have long been known for their antimicrobial increased electrostatic repulsion between the bacteria and
activity and for the capacity to interfere with the adhesion the biosurfactant-coated pellicles (Van Hoogmoed et al.
and formation of biofilms of pathogens to epithelial cells of 2006).
urogenital and intestinal tracts (Reid et al. 1998, 2001),
catheter materials (Hawthorn and Reid 1990) and voice Other biomedical and therapeutic applications
prostheses (Rodrigues et al. 2004, 2006b), and the
mechanisms of this interference have been demonstrated Biosurfactants have been shown to have many other roles in
to include, among others, the release of biosurfactants biomedical application. Surfactin is one of the most powerful
(Velraeds et al. 1996; Rodrigues et al. 2006d; Gudiña et al. biosurfactants and is known to have anti-inflammatory,
2010). Probiotics are thus well known to have a positive antibiotic and anti-tumour functions (Seydlová and Svobodová
effect on the maintenance of human health (Reid and 2008). Cao et al. (2010) demonstrated that surfactin induces
Burton 2002; Merk et al. 2005; Gupta and Garg 2009). apoptosis in human breast cancer MCF-7 cells through a
Recent work by Walencka et al. (2008) demonstrated ROS/JNK-mediated mitochondrial/caspase pathway, whereas
that surfactants obtained from three Lactobacillus acid- Byeon et al. (2008) observed that surfactin was able to down-
ophilus strains inhibited Staphylococcus epidermidis and S. regulate LPS-induced NO production in RAW264.7 cells and
primary macrophages by inhibiting NF-κB activation. Park osomes. Their results suggested that MEL-A enhanced the
and Kim (2009) studied the role of surfactin in the inhibition association of lipoplexes with the cells, delivered them
of the immunostimulatory function of macrophages through widely into the cytoplasm and increased gene expression.
blocking the NK-κB, MAPK and Akt pathway. This Ueno et al. (2007a) observed that MEL-A-containing
provided a new insight into the immunopharmacological role liposomes exhibited high activity in DNA capsulation and
of surfactin in autoimmune disease and transplantation. Their membrane fusion with anionic liposomes, which are
work indicated that surfactin has potent immunosuppressive important properties for gene transfection. On the other
capabilities which suggested important therapeutic implica- hand, MEL-B- and MEL-C-containing liposomes only
tions for transplantation and autoimmune diseases, including increased either the capsulation or the membrane fusion.
allergy, arthritis and diabetes. In another work, Ueno et al. (2007b) suggested that MEL-A
Selvam et al. (2009) studied the effect of B. subtilis PB6, was capable of increasing and rapidly promoting the
a natural probiotic, on plasma cytokine levels in inflamma- transfection efficiency of target cells by inducing membrane
tory bowel disease and colon mucosal inflammation. The fusion between liposomes and the plasma membrane of
strain was found to secrete surfactins which are known to these cells.
inhibit phospholipase A2, involved in the pathophysiology In another work, a liposome vector containing beta-
of inflammatory bowel disease. In animal experiments sitosterol beta-D-glucoside biosurfactant-complexed DNA
carried out in rat models for trinitrobenzene sulfonic acid- was successfully used for herpes simplex virus thymidine
induced colitis, oral administration of PB6 as a probiotic kinase gene therapy (Maitani et al. 2006). More recently,
suppressed colitis as measured by mortality rate and nano-vectors containing a biosurfactant have been success-
changes in colon morphology and weight gain. Plasma fully used to increase the efficacy for gene transfection in
levels of pro-inflammatory cytokines were also significant- vitro and in vivo (Nakanishi et al. 2009). On the other hand,
ly lowered and the anti-inflammatory cytokine significantly Morita et al. (2009), using a three-dimensional cultured
increased after the oral administration of PB6, supporting human skin model, observed that the viability of the SDS-
the concept that PB6 inhibits PLA2 by secreting surfactins. damaged cells was markedly improved by the addition of
Han et al. 2008 observed that high surfactin micelle MEL-A in a dose-dependent manner. This demonstrated
concentration affected the aggregation of amyloid β-peptide that MEL-A had a ceramide-like moisturising activity
(Aβ (1-40)) into fibrils, a key pathological process toward the skin cells.
associated with Alzheimer’s disease. Another interesting Another interesting application for natural surfactant is
property of surfactin and its synthetic analogues is the the possibility to synthesise metal-bound nanoparticles
ability to alter the nanoscale organisation of supported using an environmentally friendly technology benign
bilayers and to induce nanoripple structures with intriguing (Palanisamy and Raichur 2009). The use of gold nano-
perspectives for biomedical and biotechnological applica- particles, in particular, is currently undergoing a dramatic
tions (Bouffioux et al. 2007; Brasseur et al. 2007; Francius expansion in the field of drug and gene delivery, targeted
et al. 2008). Fengycin, another lipopeptide biosurfactant is therapy and imaging (Pissuwan et al. 2009; Boisselier and
also able to cause membrane perturbations (Deleu et al. Astruc 2009). Recently, Reddy et al. (2009) synthesised, for
2008). Recent results by Eeman et al. (2009) emphasised the first time, surfactin-mediated gold nanoparticles, open-
the ability of fengycin to interact with the lipid constituents ing the way to a new and fascinating application of
of the stratum corneum extracellular matrix and with biosurfactants in the biomedical field. Most recently Smyth
cholesterol. et al. (2010c) reported on the production of selectively
The biological activities and the numerous potential deuterated rhamnolipids and sophorolipids using deuterated
applications of mannosylerythritol lipids (MELs), one of substrates. The production of such deuterated biosurfactants
the most promising glycolipid biosurfactants produced by in particular or other bioactive microbial products in
yeast strains of the genus Pseudozyma, have been thor- general in which distinct pattern of labelling could be
oughly discussed by Kitamoto et al. (2009). Imura et al. achieved would have great future implications with regards
(2007) and (2008), Ito et al. (2007) and Konishi et al. to efficacy and/or persistence or the development of
(2007a) developed and studied the kinetics of interactions resistance for some bioactives particularly in biomedical
in carbohydrate ligand systems composed of self-assembled related applications.
monolayers of mannosylerythritol lipid-A (MEL-A) from P.
antarctica serving as a high-affinity, easy to handle and
low-cost ligand system for immunoglobulin G and M and Production and optimisation
lectins. Igarashi et al. (2006) reported that MEL-A
significantly increased the cellular association and the Despite their environmentally favourable characteristics of
efficiency of gene transfection mediated by cationic lip- higher biodegradability, lower toxicity, better foaming
properties compared to their synthetic chemical counter- costly or demanding downstream processing. Novel
parts while also showing better stability at extreme pH, strains able to produce biosurfactants on renewable and
salinity and temperature, the commercialisation of microbi- low-cost substrates have also been reported during the past
al surfactants has not been fully achieved largely due to few years. Ruggeri et al. (2009) isolated Rhodococcus sp.
production costs. At present, the production costs for most BS32 able to grow on rapeseed oil for the production of
biosurfactants do not compete with those of chemical extracellular biosurfactants. Glycerol, however, has
surfactants. Different strategies have been proposed to emerged as one important potential feedstock available
make the process more cost effective including: (1) in large quantities as a by-product of the biodiesel process
development of more efficient bioprocesses, including (Zheng et al. 2008).
optimisation of fermentative conditions and downstream Experimental design techniques have been extensively
recovery processes, (2) use of cheap and waste substrates used to optimise biosurfactant production. The use of
(Thavasi et al. 2007, 2008; Raza et al. 2009), (3) surface response methodology effectively enhanced the
development of overproducing strains (Fig. 3). The increas- production of biosurfactant by Rhodococcus spp. MTCC
ing number of reports of potential antimicrobial and anti- 2574 growing on n-hexadecane with yields of biosurfactant
adhesive properties of biosurfactants against pathogenic increasing from 3.2 to 10.9 g/L (Mutalik et al. 2008).
microorganisms (Rodrigues et al. 2006a) has added to the Working with Gordonia sp. BS29, Franzetti et al. (2009a)
impetus towards sustainability and reduced carbon foot increased the production of cell-bound glycolipids by 5-
prints (the greening process) which are helping drive the fold using surface response methodology, while the use of
market towards efficient large-scale production technolo- an artificial neutral network coupled with a genetic
gies. However, most biosurfactant research related to large- algorithm gave a 3.5-fold enhancement in biosurfactant
scale economic production trials has been mainly confined yield (Pal et al. 2009). The same methodologies were
to microorganisms, such as Pseudomonas, Bacillus and applied by Sivapathasekaran et al. (2010) aimed at
Candida (Mukherjee et al. 2006). optimising biosurfactant production by B. circulans MTCC
Several developments in optimisation of culture condi- 8281. Kronemberger et al. (2008) reported a significant
tions and downstream processing have been published increase in yield by optimising cultural conditions using
recently. The use of agroindustrial by-products has been statistical tools. They also reported that the yields may be
reported both for yeasts and bacteria (Makkar and Cameotra further enhanced by the development of new controlling
2002). Sobrinho et al. (2008) used ground nut oil refinery devices such as oxygen control.
residues and corn steep liquor as substrates for anionic Downstream processing accounts for most of the total
glycolipid production by Candida spherical, while the cost of a biotechnology product (Mukherjee et al. 2006).
biosynthesis of glycolipids by P. aeruginosa was obtained The most common isolation techniques for biosurfactants
using cashew apple juice as substrate (Rocha et al. 2007) use precipitation, solvent extraction and chromatographic
and vegetable oil refinery wastes (Raza et al. 2007). Very purification. Extraction of low-molecular-weight biosur-
high potential for large-scale industrial application was factant normally involves an optional precipitation step
achieved using the already commercialised Pharmamedia and the use of different organic solvents according to
medium for surfactin production by B. subtilis MZ-7 (Al- hydrophobicity and Hydrophilic-Lipophilic Balance
Ajiani et al. 2007). The use of substrates such as (HLB) value of the compounds. Rhamnolipids are usually
soapstick, frying oil and motor oil have all been explored precipitated by acidification and extracted using ethyl
and have had limited success due to the need for more acetate; extraction of sophorolipids is normally carried out
Fig. 3 Different cost-reduction

strategies for production of
microbial surface-active
compounds
with n-hexane, while for trehalolipids, the preferred non-pathogenic P. putida (Cabrera-Valladares et al. 2006;
solvent is a mixture of chloroform and methanol. Cha et al. 2008). Wang et al. (2007) also reported the use of
Methanol is also used as a solvent for extraction of genetic engineering to obtain an E. coli and P. aeruginosa
lipopeptides after a precipitation by acidification (Smyth that were able to produce rhamnolipids after transposon-
et al. 2010a). High-molecular-weight biosurfactants are mediated chromosome integration of the rhamnosyltrasferase
usually extracted from the culture broth by ammonium 1 complex. Further yield increase could probably be obtained
sulphate precipitation and purified by dialysis. Other once the regulation mechanism of biosurfactant production is
techniques for high-molecular-weight biosurfactant isola- fully elucidated (Hsueh et al. 2007).
tion are TCA/acetone precipitation, acid ethanol and
chloroform/methanol (Smyth et al. 2010b). These techni-
ques are already well established for lab-scale applica- Conclusions and perspectives
tions, but their cost does not allow scaling-up for
industrial production of biosurfactants. For these reasons, In bioremediation, biosurfactant applications are limited even
the research effort is now directed towards the develop- though their high potential has been already demonstrated.
ment of low-cost extraction and purification procedures, This field will probably benefit more from recent and future
avoiding the use of hazardous and costly organic solvents. research on the mechanisms of interactions among hydro-
Many advances have been observed in very recent years carbons, surfactants and cells than from case-specific studies
for recovery and purification of lipopeptides (Smyth et al. about applicability of already-known biosurfactant com-
2010a, b). Sivapathasekaran et al. (2009) developed and pounds. In the food, biomedical and cosmetics area, in
optimised an efficient method for the separation and which high-value products are produced, the cost draw-
purification of fengycin isoforms using high-performance back could be less significant. The complex mixture of
liquid chromatography through manipulating the solvent different components produced by organisms hampers
gradient program and flow rates. Fengycin separation and applications, and further research is required to resolve
purification was also obtained directly from the cultivation specific issues. This field will have benefits from the very
step without the use of solvent and foam formation by recent attention paid to the isolation and characterisation of
pressing and harvesting the liquid surface layer (Glazyrina biosurfactants produced by extremophiles such as thermo-
et al. 2008). Dimitrov et al. (2008) applied liquid membrane philic and halophilic bacteria (Mnif et al. 2009, Joshi et al.
extraction processes for recovery of surfactin, achieving 2008; Kumar et al. 2008) aimed at the isolation of new
97% efficiency under optimised conditions. Chen and compounds with novel properties.
colleagues, in three different papers (Chen et al. 2008a, b; The proven antimicrobial, anti-adhesive, immune-
Chen and Juang 2008), optimised the recovery of surfactin modulating properties of biosurfactants and the recent
from fermentation broths of B. subtilis ATCC 21332 by successful applications in gene therapy, immunotherapy and
different methods and achieved improved purity by medical insertion safety suggest that it is worth persisting in
adsorption or ion exchange after the broth had been treated this field. Advances in the area of biomedical application are
by a two-stage ultrafiltration process. probably going to take the lead due to higher potential
With regards to the development of over producer economic returns. Moreover, due to their self-assembly
strains, genetic manipulation of selected strains remains properties, new and fascinating applications in nanotechnol-
limited. Although recombinant strains of Bacillus sp. and ogy are predicted for biosurfactants (Palanisamy 2008; Reddy
Acinetobacter sp. have been described, most genetic et al. 2009, Kitamoto et al. 2009). In-depth studies of their
manipulation efforts have been directed towards P. aerugi- natural roles in microbial competitive interactions, cell-to-cell
nosa due in part to its commercial potential and the more communication, pathogenesis, motility and biofilm formation
detailed knowledge of its genome. Random mutagenesis and maintenance could suggest future improved and interest-
using gamma-ray or N-methyl-N’-nitrosoguanidine in- ing applications.
creased rhamnolipid production two- to threefold compared The commercial success of microbial surfactants is
to wild strains (Mukherjee et al. 2006). currently limited by the high cost of production. Optimised
The ability to produce a hyper-producer strain of P. growth/production conditions using cheaper renewable
aeruginosa, however, is quite a difficult task due to the substrates and novel and efficient multi-step downstream
complexity of the transcriptional regulatory network of genes processing methods could make biosurfactant production
involved in rhamnolipid production. This is further compli- more profitable and economically feasible. Furthermore,
cated by the fact that rhamnolipids are produced as a mixture recombinant and mutant hyper-producer microbial strains,
of congeners. Attempts have been made to limit the products able to grow on a wide range of cheap substrates, could
to mono-rhamnolipids only through cloning the P. aerugi- produce biosurfactants in high yield and, potentially, bring
nosa rhlABRI operon into host organisms such as E. coli or the required breakthrough for their economic production.
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Banat 2010

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Appl Microbiol Biotechnol (2010) 87:427–444

Microbial biosurfactants production, applications

Abstract Microorganisms synthesise a wide range of Keywords Biosurfactants . Bioemulsifiers . Surfactants .

Acidic Sophorolipid Lactonic Sophorolipid

charides, combined with long-chain aliphatic acids or Biosurfactant-enhanced bioremediation

uncertainty than biosurfactant-enhanced bioremediation, stimulate biosurfactant production in situ by indigenous

Antimicrobial activity of biosurfactants against methicillin and vancomycin) in tested strains. In

Fig. 2 Anti-adhesion activity of

Fig. 3 Different cost-reduction

References surfactin-phospholipid monolayers: effect of phospholipid chain

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