You are on page 1of 8

Trends in Food Science & Technology 18 (2007) 252e259

Review

Biosurfactants in
food industry Biosurfactants (bioemulsifiers) are a structurally diverse
group of surface-active molecules synthesized by microor-
ganisms. Rhamnolipids from Pseudomonas aeruginosa,
M. Nitschkea,* surfactin from Bacillus subtilis, emulsan from Acineto-
bacter calcoaceticus and sophorolipids from Candida
and S.G.V.A.O. Costab bombicola are some examples of microbial-derived surfac-
a
tants. Originally, biosurfactants attracted attention as hydro-
Department of Microbiology, carbons dissolution agents, but the interest in these
EMBRAPA Food Technology, Av. das Américas, molecules have been increasing considerably in the past
29501, Rio de Janeiro, RJ CEP 23020-470, Brazil five decades as alternative to chemical surfactants (carboxy-
(Tel.: D55 21 24109590; fax: D55 21 24101090; lates, sulphonates and sulphate acid esters) specially in
e-mail: nitschke@ctaa.embrapa.br) food, pharmaceutical and oil industry (Banat, Makkar, &
b
Department of Biochemistry and Microbiology, Cameotra, 2000; Desai & Banat, 1997). The main reasons
Institute of Biological Sciences, for the spreading interest in biosurfactants are their environ-
UNESP/Rio Claro, Av. 24-A, 1515, mental friendly nature, since they are easily biodegradable
CEP 13506-900, C.Postal 199, (Mohan, Nakhla, & Yanful, 2006) and have low toxicity
Rio Claro- SP, Brazil (Flasz, Rocha, Mosquera, & Sajo, 1998), and their unique
structures which provide new properties that classical
surfactants may lack.
The increasing environmental concern about chemical surfac-
Most work on biosurfactants applications has been
tants triggers attention to microbial-derived surface-active
focusing on bioremediation of pollutants (Mulligan, 2005);
compounds essentially due to their low toxicity and biode-
however, these microbial compounds exhibit a variety of
gradable nature. At present, biosurfactants are predominantly
useful properties for the food industry specially as emulsi-
used in remediation of pollutants; however, they show poten-
fiers, foaming, wetting, solubilizers (Banat et al., 2000),
tial applications in many sectors of food industry. Associated
antiadhesive and antimicrobial agents (Singh & Cameotra,
with emulsion forming and stabilization, antiadhesive and an-
2004). Moreover, an increasing consciousness among con-
timicrobial activities are some properties of biosurfactants,
sumers demands for reducing the use of artificial or chem-
which could be explored in food processing and formulation.
ically synthesized compounds by replacing it for more
Potential applications of microbial surfactants in food area and
natural food ingredients and additives (Shepherd, Rockey,
the use of agroindustrial wastes as alternative substrates for
Sutherland, & Roller, 1995). Despite the advantages
their production are discussed.
demonstrated by biosurfactants, few reports are available
regarding their use on food products and food processing.
This paper discusses the properties and applications of
microbial surfactants which can be of interest for food
Introduction and food-related industries, providing also an overview of
Surfactants are amphiphilic compounds containing both the emerging fields for their employment and considering
hydrophobic (nonpolar) and hydrophilic (polar) moieties the exploitation of agroindustrial wastes as alternative
that confer ability to accumulate between fluid phases substrates for biosurfactant production.
such as oil/water or air/water, reducing the surface and in-
terfacial tensions and forming emulsions (Desai & Banat,
Classification of biosurfactants
1997). The surface activity properties make surfactants
Microbial surfactants are categorized by their chemical
one of the most important and versatile class of chemical
composition and microbial origin. Rosenberg and Ron
products, used on a variety of applications in household,
(1999) suggested that biosurfactants can be divided into
industry and agriculture (Deleu & Paquot, 2004).
low-molecular-mass molecules, which efficiently lower
surface and interfacial tension, and high-molecular-mass
* Corresponding author. polymers, which are more effective as emulsion stabilizing
0924-2244/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tifs.2007.01.002
M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259 253

agents. The major classes of low-mass surfactants include Surfactin from B. subtilis can reduce ST of water to
glycolipids, lipopeptides and phospholipids, whereas high 25 mN/m and IT water/hexadecane to <1 mN/m (Cooper,
mass includes polymeric and particulate surfactants. MacDonald, Duff, & Kosaric, 1981). The rhamnolipids
Most biosurfactants are either anionic or neutral and the from P. aeruginosa decreased ST of water to 26 mN/m
hydrophobic moiety is based on long-chain fatty acids or and IT water/hexadecane to value <1 mN/m (Syldatk,
fatty acids derivatives whereas the hydrophilic portion Lang, & Wagner, 1985) however, some rhamnolipid homo-
can be a carbohydrate, aminoacid, phosphate or cyclic pep- logues have demonstrated lower values (Nitschke, Costa, &
tide. Table 1 shows the major biosurfactant classes and the Contiero, 2005). The sophorolipids from C. bombicola
microorganisms involved; note that bacteria are the pre- were reported to reduce ST to 33 mN/m and IT to 5 mN/m
dominant group of surfactant-producing organisms how- (Cooper & Paddock, 1984). In general, biosurfactants are
ever some yeast species are also involved. Bodour et al. more effective and efficient and their CMC (critical micelle
(2004) have recently described a new class of biosur- concentration) is about 10e40 times lower than chemical
factants named flavolipids produced by a soil isolated surfactants, i.e., less surfactant is necessary to get a maximal
Flavobacterium sp. The new surfactant showed strong decrease on ST (Desai & Banat, 1997).
surface activity and emulsifying ability, and exhibits a polar
moiety that features citric acid. Temperature, pH and ionic strength tolerance
Many biosurfactants and their surface activity are not af-
Properties of biosurfactants fected by environmental conditions such as temperature and
The main distinctive features of microbial surfactants pH. McInerney, Javaheri, and Nagle (1990) reported that li-
that can be of interest for food processing are related to chenysin from Bacillus licheniformis JF-2 was not affected
their surface activity; tolerance to pH, temperature and by temperature (up to 50  C), pH (4.5e9.0) and by NaCl
ionic strength; biodegradability; low toxicity; emulsifying and Ca concentrations up to 50 and 25 g/L, respectively.
and demulsifying ability and antimicrobial activity. These A lipopeptide from B. subtilis LB5a was stable after auto-
topics are discussed below. clave (121  C/20 min) and after 6 months at 18  C; the
surface activity did not change from pH 5 to pH 11 and
Surface and interface activity NaCl concentrations up to 20% (Nitschke & Pastore,
A good surfactant can lower surface tension (ST) of 2006). The Antarctic psychrophilic strain Arthrobacter pro-
water from 72 to 35 mN/m and the interfacial tension (IT) tophormiae produced a biosurfactant that was thermostable
water/hexadecane from 40 to 1 mN/m (Mulligan, 2005). (30e100  C) and pH (2e12) stable (Pruthi & Cameotra,
1997). Industrial processes frequently involve exposure to
extremes of temperature, pressure, pH and ionic strength,
Table 1. Major types of microbial surfactants hence there is a continuous need to isolate new microbe-
Surfactant class Microorganism derived products able to function under these conditions
Glycolipids (Cameotra & Makkar, 1998). The production of surfactants
Rhamnolipids Pseudomonas aeruginosa from extremophiles microorganisms, has gained attention
Trehalose lipids Rhodococcus erithropolis, in last years once the unique properties of these compounds
Arthobacter sp. are of considered commercial interest.
Sophorolipids Candida bombicola, Candida
apicola
Mannosylerythritol lipids Candida antartica Biodegradability
Lipopeptides Unlike synthetic surfactants, microbial-produced com-
Surfactin/iturin/fengycin Bacillus subtilis pounds are easily degraded (Mohan et al., 2006) and partic-
Viscosin Pseudomonas fluorescens ularly suited for environmental applications such as
Lichenysin Bacillus licheniformis
bioremediation (Deleu & Paquot, 2004; Mulligan, 2005).
Serrawettin Serratia marcescens
Phospholipids Acinetobacter sp., The increasing environmental concern among consumers
Corynebacterium lepus and the regulatory rules imposed by governments forcing
Fatty acids/neutral lipids industry to search for alternative products such as biosur-
Corynomicolic acids Corynebacterium insidibasseosum factants (Cameotra & Makkar, 1998).
Polymeric surfactants
Emulsan Acinetobacter calcoaceticus
Alasan Acinetobacter radioresistens Low toxicity
Liposan Candida lipolytica Although few data are available in literature regarding
Lipomanan Candida tropicalis the toxicity of microbial surfactants, they are generally con-
Particulate biosurfactants sidered low or non-toxic products and therefore, appropri-
Vesicles Acinetobacter calcoaceticus
ate for pharmaceutical, cosmetic and food uses. Poremba,
Whole microbial cells Cyanobacteria
Gunkel, Lang, and Wagner (1991a) reported that a synthetic
Adapted from Deleu and Paquot, 2004; Desai and Banat, 1997; anionic surfactant (Corexit) displayed a LC50 (concentra-
and Rosenberg and Ron, 1999.
tion lethal to 50% of test species) against Photobacterium
254 M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259

phosphoreum 10 times lower than rhamnolipids, demon- but not to be good emulsifiers (Cooper & Paddock,
strating the higher toxicity of the chemical-derived surfac- 1984). By contrast, liposan has been shown not to reduce
tant. When comparing the toxicity of six biosurfactants, surface tension but used successfully to emulsify edible
four synthetic surfactants and two commercial dispersants, oils (Cirigliano & Carman, 1985). Polymeric surfactants
Poremba, Gunkel, Lang, and Wagner (1991b) found that offer additional advantages because they coat the droplets
most biosurfactants were degraded faster, except for a syn- of oil, thereby forming very stable emulsions that never
thetic sucrose-stearate that showed structure homology to coalesce. This property is especially useful for making oil/
glycolipids and was degraded more rapidly than the bio- water emulsions for cosmetics and food. In dairy products
genic glycolipids (rhamnolipids, trehalose lipids, sophorose (soft cheese and ice creams) the addition of emulsifiers im-
lipids). These authors also reported that biosurfactants proves the texture and creaminess. This quality is of special
showed higher EC50 (effective concentration to decrease value for low-fat products (Rosenberg & Ron, 1999).
50% of test population) values than synthetic dispersants. Evaluation of emulsifying ability of biosurfactants is in
A biosurfactant from P. aeruginosa was compared with general related to hydrocarbons such as kerosene because
a synthetic surfactant (Marlon A-350) widely used in indus- of their potential in environmental applications. Few at-
try in terms of toxicity and mutagenic properties. Both as- tempts have been made to evaluate emulsion forming
says indicated the higher toxicity and mutagenic effect of by biosurfactants with oils and fats used in food industry.
the chemical-derived surfactant whereas biosurfactant was A lipopeptide obtained from B. subtilis was able to form
considered slightly to non-toxic and non-mutagenic (Flasz stable emulsions with soybean oil and coconut fat, suggest-
et al., 1998). The comparison of acute and chronic toxicity ing its potential as emulsifying agent in foods (Nitschke &
of three synthetic surfactants (Corexit, 9500, Triton X-100, Pastore, 2006).
PSE-61) and three microbiological derived surfactants A manoprotein from Kluyveromyces marxianus was able
(rhamnolipid, emulsan, biological cleanser PES-51) com- to form emulsions with corn oil that were stable for 3
monly used in oil spill remediation revealed that PES-61 months; the yeast was cultivated on whey-based medium
(synthetic surfactant) and Emulsan (biosurfactant) were suggesting potential application as food bioemulsifier
the least toxic whereas Triton X-100 (synthetic) was the (Lukondeh, Ashbolh, & Rogers, 2003). The extracellular
most toxic (Edwards, Lepo, & Lewis, 2003). carbohydrate-rich compound from Candida utilis was
Rhamnolipid surfactants are presently produced at successfully used as emulsifying agent in salad dressing
commercial scale by Jeneil Biosurfactant Corp. (www. formulations (Shepherd et al., 1995). The use of yeast for
biosurfactant.com) which offers diverse formulations for production of biosurfactant is interesting because these or-
different purposes. Recently, they developed a biofungicide ganisms are generally recognized as safe (GRAS) and they
formulation to prevent plant pathogenic fungi that was are already present in many food manufacturing processes,
considered of low acute mammalian toxicity and non- on the contrary, products derived from bacteria such as the
mutagenic and was approved by FDA for use in fruit, opportunistic P. aeruginosa, still face some resistance con-
vegetables and legume crops. Additionally, the greater con- cerning their use as food ingredients.
sumer awareness of adverse allergic effects caused by arti- In some case, the emulsion, which is generated in one
ficial products stimulates the development of alternative part of the process, may have to be destabilized in a sub-
ingredients, thus opening an excellent opportunity to sequent operation to develop a certain functional property
expand the use of natural surfactants of microbial origin to the final product. De-emulsification can be of interest
(Cameotra & Makkar, 1998). in food processing specially when related to fat and oil
products as well as in waste treatment (Kachholz &
Emulsion forming and emulsion breaking Schlingmann, 1987).
An emulsion is a heterogeneous system, consisting of at
least one immiscible liquid intimately dispersed in another Antimicrobial activity
in the form of droplets, whose diameter in general exceeds Several biosurfactants have shown antimicrobial action
0.1 mm. Emulsions have an internal or dispersed and an ex- against bacteria, fungi, algae and viruses. The lipopeptide
ternal or continuous phase, so there are generally two types: iturin from B. subtilis showed potent antifungal activity
oil-in-water (o/w) or water-in-oil (w/o) emulsions. Such (Besson, Peypoux, Michel, & Delcambe, 1976). A signifi-
systems possess a minimal stability, which may be accentu- cative reduction on the mycoflora present in stored grains
ated by additives such as surface-active agents (surfac- of corn, peanuts and cottonseeds was observed at iturin
tants). Thus, stable emulsions can be produced with a life concentration of 50e100 ppm (Klich, Arthur, Lax, &
span of months and years (Velikonja & Kosaric, 1993). Bland, 1994). Inactivation of enveloped virus such as
Biosurfactants may stabilize (emulsifiers) or destabilize herpes and retrovirus was observed with 80 mM of sur-
(de-emulsifiers) the emulsion. High-molecular-mass biosur- factin (Vollenbroich, Özel, Vater, Kamp, & Pauli, 1997).
factants are in general better emulsifiers than low-molecular- Rhamnolipids inhibited the growth of harmful bloom
mass biosurfactants. Sophorolipids from Torulopsis bombicola algae species Heterosigma akashivo and Protocentrum
have been shown to reduce surface and interfacial tension dentatum at concentration ranging from 0.4 to 10.0 mg/L
M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259 255

(Wang et al., 2005). A rhamnolipid mixture obtained from L-Rhamnose has a considerable potential as precursor
P. aeruginosa AT10 showed inhibitory activity against the for flavorings. It is already used industrially as precursor
bacteria Escherichia coli, Micrococcus luteus, Alcaligenes of high-quality flavor components like Furaneol (trademark
faecalis (32 mg/mL), Serratia marcescens, Mycobacterium of Firmenich SA, Geneva). There is great interest in obtain-
phlei (16 mg/mL) and Staphylococcus epidermidis (8 mg/ ing rhamnose lipids to provide a source of L-rhamnose,
mL); excellent antifungal properties against Aspergillus which already has an industrial application. L-Rhamnose
niger (16 mg/mL), Chaetonium globosum, Penicillium is obtained by hydrolyzing rhamnolipid surfactants pro-
crysogenum, Aureobasidium pullulans (32 mg/mL) and the duced by P. aeruginosa (Linhardt, Bakhit, Daniels, Mayerl,
phytopathogenic Botrytis cinerea and Rhizoctonia solani & Pickenhagen, 1989).
(18 mg/mL) (Abalos et al., 2001). Sophorolipids and rham-
nolipids were found to be effective antifungal agents Antiadhesive agents
against plant and seed pathogenic fungi. Mycelial growth A biofilm is described as a group of bacteria that have
of Phytophthora sp. and Pythium sp. was 80% inhibited colonized a surface. The biofilm not only includes the bac-
by 200 mg/L of rhamnolipids and 500 mg/L of sophoro- teria, but it also describes all of the extracellular material
lipids (Yoo, Lee, & Kim, 2005). The mannosylerythritol produced at the surface and any material trapped within
lipid (MEL), a glycolipid surfactant from Candida antar- the resulting matrix (Hood & Zottola, 1995). The first
tica has demonstrated antimicrobial activity particularly step on biofilm establishment is bacterial adherence which
against Gram-positive bacteria (Kitamoto et al., 1993). Be- is affected by factors including microorganism species,
sides their antimicrobial activity new biological applica- hydrophobicity of surface and electrical charges involved,
tions of biosurfactants have been found and some reviews environmental conditions and ability of microorganisms
concerning the potential uses of microbial surfactants in to produce extracellular polymers that help cells to anchor
biomedical sciences were recently published (Rodrigues, to surfaces (Zottola, 1994). Bacterial biofilms present in
Banat, Teixeira, & Oliveira, 2006; Singh & Cameotra, food industry surfaces are potential sources of contamina-
2004). tion, which may lead to food spoilage and disease transmis-
sion (Hood & Zottola, 1995). Due to the fact that food
Potential food applications processors have a zero tolerance levels for pathogens like
Although biosurfactants can be explored for several food Salmonella and also (in most countries) for Listeria mono-
processing applications, in this section we emphasize their cytogenes, a single adherent cell may be as significant as
potential as food formulation ingredients and antiadhesive a well developed biofilm; thus controlling the adherence
agents. of microorganisms to food contact surfaces is an essential
step in providing safe and quality products to consumers
Food formulation ingredients (Hood & Zottola, 1995).
Apart from their obvious role as agents that decrease The involvement of biosurfactants in microbial adhesion
surface and interfacial tension, thus promoting the forma- and detachment from surfaces has been investigated. A sur-
tion and stabilization of emulsions, surfactants can have factant released by Streptococcus thermophilus has been
several other functions in food. For example to control used for fouling control of heat-exchanger plates in pasteur-
the agglomeration of fat globules, stabilize aerated systems, izers as it retards the colonization of other thermophilic
improve texture and shelf-life of starch-containing prod- strains of Streptococcus responsible for fouling (Busscher,
ucts, modify rheological properties of wheat dough and van der Kuij-Booij, & van der Mei, 1996).
improve consistency and texture of fat-based products The bioconditioning of surfaces through the use of micro-
(Kachholz & Schlingmann, 1987). bial surfactants have been suggested as a new strategy to
In bakery and ice cream formulations biosurfactants act reduce adhesion. Pre-treatment of silicone rubber with
controlling consistency, retarding staling and solubilizing S. thermophilus surfactant inhibited by 85% the adhesion of
flavor oils; they are also utilized as fat stabilizer and anti- Candida albicans (Busscher, van Hoogmoed, Geertsema-
spattering agent during cooking of oil and fats (Kosaric, Doornbusch, van der Kuij-Booij, & van der Mei, 1997)
2001). An improvement of dough stability, texture, volume whereas surfactants from Lactobacillus fermentum and Lac-
and conservation of bakery products was obtained by the tobacillus acidophilus adsorbed on glass, reduced by 77%
addition of rhamnolipid surfactants (Van Haesendonck & the number of adhering uropathogenic cells of Enterococcus
Vanzeveren, 2004). The authors also suggested the use of faecalis (Velraeds, van der Mei, Reid, & Busscher, 1996).
rhamnolipids to improve properties of butter cream, crois- Lately, the biosurfactant from L. fermentum was reported
sants and frozen confectionery products. Recently, a bio- to inhibit Staphylococcus aureus infection and adherence
emulsifier isolated from a marine strain of Enterobacter to surgical implants (Gan, Kim, Reid, Cadieux, & Howard,
cloacae was described as a potential viscosity enhancement 2002). The use of biosurfactants released by Lactobacilli
agent of interest in food industry especially due to the good strains is very promising once these microorganisms are nat-
viscosity observed at acidic pH allowing its use in products urally present in human flora and have also a probiotic effect
containing citric or ascorbic acid (Iyer, Mody, & Jha, 2006). (Singh & Cameotra, 2004). Much more research is needed
256 M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259

however, to understand the contribution of lactobacilli sur- Considering the interesting properties demonstrated by
factants in preventing pathogen colonization, the biochemi- biosurfactants we can think on their future utilization as
cal aspects of biosynthesis and their structural characterization. multipurpose ingredients, which exhibit emulsifier, anti-
Surfactin decreased the amount of biofilm formation by adhesive, and antimicrobial activities simultaneously and
Salmonella typhimurium, Salmonella enterica, E. coli and thus, suitable for many food applications. Food processors
Proteus mirabilis in PVC plates and vinyl urethral catheters however, does not yet use biosurfactants on a large scale as
(Mireles, Toguchi, & Harshey, 2001). Irie, o’Toole, and many regulations regarding the approval of new food ingre-
Yuk (2005) demonstrated the disruption of Bordetella bron- dients are required by governmental agencies, and this pro-
chiseptica biofilms by rhamnolipids and recently, it was cess could be quite long. Nevertheless, an increasing
found that silicone rubber conditioned with rhamnolipids number of patents have been issued on biosurfactant (bio-
reduced the adhesion rates of Streptococcus salivarius and emulsifier) claiming their use as additives for food, cos-
Candida tropicalis by 66%. The number of adhered cells metics and pharmaceutical products (Shete, Wadhawa,
of S. aureus, S. epidermidis, S. salivarius and C. tropicalis Banat, & Chopade, 2006), demonstrating the crescent inter-
was reduced by 48% and the perfusion of biosurfactant to est in using these microbial-derived products.
adhered cells produced a high detachment (96%) of micro-
organisms (Rodrigues, Banat, van der Mei, Teixeira, & Biosurfactant production from food and
Oliveira, 2006). agroindustrial wastes
The use of biosurfactants, which disrupts biofilms and Another interesting approach for food industries, is to
reduce adhesion, in combination with antibiotics could rep- take advantage of their by-products or residues as substrates
resent a novel antimicrobial strategy, once antibiotics are in for biosurfactant production. Currently, the main factor that
general less effective against biofilms than planktonic cells; works against the widespread use of biosurfactants is the
the disruption of biofilm by biosurfactant can facilitate the economics of their production (Makkar & Cameotra, 2002).
antibiotic access to the cells (Irie et al., 2005). An important point that should be considered for develop-
The promising results of these works, with medical focus, ment of cheaper processes is the selection of inexpensive
suggest a potential application of biosurfactants for surface medium components which account for 10e30% of overall
conditioning in food industry, since both the surface costs (Cameotra & Makkar, 1998). To this end, good com-
materials and microorganisms involved are of common ponents seem to be agroindustrial by-products or wastes,
interest. once these residues generally contain high levels of carbo-
An interesting work regarding the use of biosurfactants hydrates or lipids to support growth and surfactant synthe-
to inhibit the adhesion of the pathogen L. monocytogenes sis; moreover, the treatment and disposal costs for these
in two types of surfaces classically used in food industry residues are significant to industries which are invariably
has been conducted by the group of Meylheuc, van Oss, searching for alternatives to reduce, reuse, recycle and
and Bellon-Fontaine (2001). The preconditioning of stain- valorize their wastes.
less steel and PTFE surfaces with a biosurfactant obtained The advantages of wastes materials are the huge sur-
from Pseudomonas fluorescens inhibits the adhesion of pluses and the possibility of production in regions with
L. monocytogenes L028 strain. A significant reduction temperate to tropical climates (Mercade & Manresa,
(>90%) was attained in microbial adhesion levels in stain- 1994). Thus, the research on alternative low-cost substrates
less steel whereas no significant effect was observed in is mainly centered on tropical agroindustrial crops and res-
PTFE. Further work demonstrated that the prior adsorp- idues. Some attempts at using wastes for biosurfactant pro-
tion of P. fluorescens surfactant in stainless steel also fa- duction and only a few types of surfactant produced from
vored the bactericidal effect of disinfectants (Meylheuc, wastes have been published. The main alternative sources
Renault, & Bellon-Fontaine, 2006). The ability of ad- for biosurfactant production comprise oily residues, milk
sorbed biosurfactants obtained from Gram-negative and distillery wastes, and carbohydrate-rich residues.
(P. fluorescens) and Gram-positive (Lactobacillus helveti-
cus) bacteria isolated from foodstuffs, in inhibiting the ad- Oils and fats
hesion of L. monocytogenes to stainless steel was recently Most oils and fats are used in the food industry, which
investigated. Adhesion tests showed that both biosurfac- generates great quantities of wastes and so, their disposal
tants were effective by decreasing strongly the level of is a growing problem. Candida antarctica and Candida api-
contamination of the surface. The antiadhesive biologic cola synthesized surfactants (glycolipids) in a cultivation
coating reduced either the total adhering flora and the vi- medium supplemented with oil refinery waste, either with
able/cultivable adherent L. monocytogenes on stainless soapstock (5e12% v/v) or post-refinery fatty acids (from
steel surfaces (Meylheuc et al., 2006). Preliminary studies 2 to 5% v/v). The efficiency of glycolipids synthesis was
regarding the corrosion effect of P. fluorescens surfactant from 7.3 to 13.4 g/L and from 6.6 to 10.5 g/L in the me-
in stainless steel suggested that it has also a good potential dium supplemented with soapstock and post-refinery fatty
as corrosion inhibitor (Dagbert, Meylheuc, & Bellon- acids, respectively (Bednarski, Adamczak, Tomasik, &
Fontaine, 2006). Plaszczyk, 2004).
M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259 257

The meat processing industry is seeking new applica- considered the most promising strain and whey as a poten-
tions for abundantly available, inexpensive animal’s fats. tial alternative substrate.
Sophorolipid production by C. bombicola was studied as
a model of fat utilization for biosurfactant production. In Carbohydrate-rich residues
a pH controlled fermenter, 120 g/L of sophorolipid was ob- Fox and Bala (2000) demonstrated that potato process-
tained and the cells at the end of fermentation contained ing effluent was suitable alternative carbon source to gener-
37% of protein and 14% lipids (Deshpande & Daniels, ate surfactant from B. subtilis ATCC 21332. B. subtilis
1995). expresses an a-amylase which permits the utilization of
Frying oils is produced in large quantities for use both a starch-rich potato waste as substrate for biosurfactant pro-
in the food industry and at the domestic scale. Haba, Es- duction (Thompson, Fox, & Bala, 2000). Cassava waste-
puny, Busquets, and Manresa (2000) studied a screening water is a carbohydrate-rich residue generated at large
process for the selection of microorganism’s strains able amounts during the processing of cassava flour. This resi-
to grow on frying oils (sunflower and olive) and accumu- due proved to be an appropriate substrate for biosurfactant
late surface-active compounds in the culture media. P. aer- biosynthesis, providing not only bacterial growth and prod-
uginosa 47T2 was selected, showing a final production of uct accumulation but also a surfactant that has interesting
rhamnolipid of 2.7 g/L and a production yield of 0.34 g and useful properties with potential for many industrial
rhamnolipid/g substrate. Sunflower oil soapstock was as- applications (Nitschke & Pastore, 2003, 2004).
sayed as the carbon source for rhamnolipid production Molasses is a by-product of the sugar industry that is low
by P. aeruginosa LBI strain, giving a final surfactant in price compared to other conventional sugar sources like
concentration of 12 g/L in shaker and 16 g/L in bioreactor sucrose or glucose and is rich in other nutrients such as
experiments (Benincasa, Contiero, Manresa, & Moraes, minerals and vitamins (Makkar & Cameotra, 2002). Two
2002). Equally Nitschke et al. (2005) evaluated edible oil B. subtilis strains were able to produce lipopeptide surfac-
soapstocks as alternative low-cost substrates for the pro- tants using minimal medium supplemented with molasses
duction of rhamnolipids by P. aeruginosa LBI strain. as carbon source (Makkar & Cameotra, 1997). Molasses
Wastes obtained from soybean, cottonseed, babassu, palm and corn steep liquor were used as the primary carbon
and corn oil refinery were tested. The soybean soapstock and nitrogen sources for production of rhamnolipid biosur-
waste was the best substrate, generating 11.7 g/L of rham- factants by P. aeruginosa GS3; the interfacial tension of
nolipids and a production yield of 75%. Vegetable oils and culture medium against crude oil was reduced from 21 to
residues from vegetable oil refinery are among the most 0.47 mN/m (Patel & Desai, 1997).
used low-cost substrates for rhamnolipids production
(Nitschke et al., 2005). Future trends
Biosurfactants show several properties which could be
useful in many fields of food industry; recently, their anti-
Whey milk and distillery wastes adhesive activity has attracted attention as a new tool to
The disposal of cheese whey is a continuing and growing inhibit and disrupt the biofilms formed in food contact
problem to the dairy industry. A two-step batch cultivation surfaces. The combination of particular characteristics
process was developed to produce sophorolipids from whey such as emulsifying, antiadhesive and antimicrobial activi-
by C. bombicola and Cryptococcus curvatus. In the first ties presented by biosurfactants suggests potential applica-
step, C. curvatus was grown on deproteinized whey concen- tion as multipurpose ingredients or additives.
trates (DWC); the cultivation broth was disrupted with Scant information regarding toxicity, combined with
a glass bead mill and it served a medium for growth and high production costs seems to be the major cause for the
sophorolipid production by C. bombicola (Daniel, Otto, limited uses of biosurfactants in food area. However, the
Binder, Reuss, & Syldatk, 1999). use of agroindustrial wastes can reduce the biosurfactants
Dubey and Juwarkar (2001) reported biosurfactant pro- production costs as well as the waste treatment expends,
duction from synthetic medium and industrial wastes such and also renders a new alternative for food and food-related
as distillery and whey by a sludge isolate P. aeruginosa industries not only for valorizing their wastes but also to
BS2. The wastes were good substrates for growth and pro- becoming microbial surfactant producers. Biosurfactants
liferation of bacteria and biosurfactant production in distill- obtained from GRAS microorganisms like Lactobacilli
ery and whey wastes reached maximal amounts of 0.9 and and yeasts are of great promise for food and medicine
0.92 g/L, respectively, after 96 h of incubation. applications though, much more research is already required
Rodrigues, Moldes, Teixeira, and Oliveira (2006) per- on this field. The prospect of new types of surface-active
formed a screening for Lactobacillus strains able to produce compounds from microorganisms can contribute for the de-
surfactants. The acid lactic bacteria Lactobacillus casei, tection of different molecules in terms of structure and prop-
Lactobacillus rhamnosus, Lactobacillus pentosus and erties, but the toxicological aspects of new and current
Lactobacillus coryniformis torquens were selected as biosurfactants should be emphasized in order to certify the
surfactant-producing organisms with L. pentosus been safe of these compounds for food utilization.
258 M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259

With the emphasis on the construction of a sustainable produced by Pseudomonas fluorescens. Electrochimica Acta, 51,
society in harmony with the environment, the introduction 5221e5227.
Daniel, H. J., Otto, R. T., Binder, M., Reuss, M., & Syldatk, C. (1999).
of green technology in all fields of industry is one of the Production of sophorolipids of whey: development of a two-stage
most important challenges. Considering the social and tech- process with Cryptococcus curvatus ATCC 20509 and Candida
nological backgrounds, utilization of biosurfactants, which bombicola ATCC 22214 using deproteinized whey concentrates
are environmentally friendly and highly functional, have as substrates. Applied Microbiology and Biotechnology, 51,
become more and more important. 40e45.
Deleu, M., & Paquot, M. (2004). From renewable vegetables resources
to microorganisms: new trends in surfactants. Comptes Rendus
Chimie, 7, 641e646.
Acknowledgements Desai, J. D., & Banat, I. M. (1997). Microbial production of surfactants
We thank Fundaç~ao de Amparo à Pesquisa do Estado and their commercial potential. Microbiological Molecular
de S~ao Paulo (FAPESP) for scholarship and Embrapa for Reviews, 61, 47e64.
providing us the facilities for this work. Deshpande, M., & Daniels, L. (1995). Evaluation of sophorolipid
biosurfactant production by Candida bombicola using animal fat.
Bioresource Technology, 54, 143e150.
References Dubey, K., & Juwarkar, A. (2001). Distillery and curd whey wastes as
viable alternative sources for biosurfactant production. World
Abalos, A., Pinazo, A., Infante, M. R., Casals, M., Garcı́a, F., & Journal of Microbiology & Biotechnology, 17, 61e69.
Manresa, A. (2001). Physicochemical and antimicrobial properties Edwards, K. R., Lepo, J. E., & Lewis, M. A. (2003). Toxicity comparison
of new rhamnolipids produced by Pseudomonas aeruginosa of biosurfactants and synthetic surfactants used in oil spill
AT10 from soybean oil refinery wastes. Langmuir, 17(5), 1367e remediation to two estuarine species. Marine Pollution Bulletin,
1371. 46(10), 1309e1316.
Banat, I. M., Makkar, R. S., & Cameotra, S. S. (2000). Potential Flasz, A., Rocha, C. A., Mosquera, B., & Sajo, C. (1998). A
commercial applications of microbial surfactants. Applied comparative study of the toxicity of a synthetic surfactant and one
Microbiology and Biotechnology, 53, 495e508. produced by Pseudomonas aeruginosa ATCC 55925. Medical
Bednarski, W., Adamczak, M., Tomasik, J., & Plaszczyk, M. (2004). Science Research, 26(3), 181e185.
Application of oil refinery waste in the biosynthesis of glycolipids Fox, S. L., & Bala, G. A. (2000). Production of surfactant from Bacillus
by yeast. Bioresource Technology, 95, 15e18. subtilis ATCC 21332 using potato substrates. Bioresource
Benincasa, M., Contiero, J., Manresa, M. A., & Moraes, I. O. (2002). Technology, 75, 235e240.
Rhamnolipid production by Pseudomonas aeruginosa LBI growing Gan, B. S., Kim, J., Reid, G., Cadieux, P., & Howard, J. C. (2002).
on soapstock as the sole carbon source. Journal of Food Lactobacillus fermentum RC-14 inhibits Staphylococcus aureus
Engineering, 54, 283e288. infection of surgical implants in rats. Journal of Infectious Diseases,
Besson, F., Peypoux, F., Michel, G., & Delcambe, L. (1976). 185(9), 1369e1372.
Characterization of iturin A in antibiotics from various strains of Haba, E., Espuny, M. J., Busquets, M., & Manresa, A. (2000).
Bacillus subtilis. Journal of Antibiotics, 29(10), 1043e1049. Screening and production of rhamnolipids by Pseudomonas
Bodour, A. A., Guerrero-Barajas, C., Jiorle, B. V., Malcomson, M. E., aeruginosa 47T2 NCBI 40044 from waste frying oils. Journal of
Paull, A. K., Somogyi, A., et al. (2004). Structure and character- Applied Microbiology, 88, 379e387.
ization of flavolipids, a novel class of biosurfactants produced by Hood, S. K., & Zottola, E. A. (1995). Biofilms in food processing. Food
Flavobacterium sp. strain MTN11. Applied and Environmental Control, 6(1), 9e18.
Microbiology, 70(1), 114e120. Irie, Y., O’Toole, G. A., & Yuk, M. H. (2005). Pseudomonas aeruginosa
Busscher, H. J., van Hoogmoed, C. G., Geertsema-Doornbusch, G. I., rhamnolipids disperse Bordetella bronchiseptica biofilms. FEMS
van der Kuij-Booij, M., & van der Mei, H. C. (1997). Streptococcus Microbiology Letters, 250, 237e243.
thermophilus and its biosurfactants inhibit adhesion by Candida Iyer, A., Mody, K., & Jha, B. (2006). Emulsifying properties of a marine
spp. on silicone rubber. Applied and Environmental Microbiology, bacterial exopolysaccharide. Enzyme and Microbial Technology,
63, 3810e3817. 38, 220e222.
Busscher, H. J., van der Kuij-Booij, M., & van der Mei, H. C. (1996). Kachholz, T., & Schlingmann, M. (1987). Possible food and
Biosurfactants from thermophilic dairy streptococci and their agricultural applications of microbial surfactants: an assessment.
potential role in the fouling control of heat exchanger plates. In N. Kosaric, W. L. Carns, & N. C. C. Gray (Eds.), Biosurfactants
Journal of Industrial Microbiology & Biotechnology, 16(1), and biotechnology (pp. 183e210). New York: Marcel
15e21. Dekker.
Cameotra, S. S., & Makkar, R. S. (1998). Synthesis of biosurfactants in Kitamoto, D., Yanagishita, H., Shinbo, T., Nakane, T., Kamisawa, C., &
extreme conditions. Applied Microbiology and Biotechnology, 50, Nakahara, T. (1993). Surface active properties and antimicrobial
520e529. activities of mannosylerythritol lipids as biosurfactants
Cirigliano, M. C., & Carman, G. M. (1985). Purification and charac- produced by Candida antarctica. Journal of Biotechnology,
terization of liposan, a bioemulsifier from Candida lipolytica. 29, 91e96.
Applied and Environmental Microbiology, 50, 846e850. Klich, M. A., Arthur, K. S., Lax, A. R., & Bland, J. M. (1994). Iturin A:
Cooper, D. G., MacDonald, C. R., Duff, S. J. B., & Kosaric, N. (1981). a potential new fungicide for stored grains. Mycopathologia,
Enhanced production of surfactin from B. subtilis by continuous 127(2), 123e127.
product removal and metal cation additions. Applied and Kosaric, N. (2001). Biosurfactants and their application for soil bio-
Environmental Microbiology, 42, 408e412. remediation. Food Technology and Biotechnology, 39(4), 295e304.
Cooper, D. G., & Paddock, D. A. (1984). Production of a biosurfactant Linhardt, R. J., Bakhit, R., Daniels, L., Mayerl, F., & Pickenhagen, W.
from Torulopsis bombicola. Applied and Environmental (1989). Microbially produced rhamnolipid as a source of
Microbiology, 47, 173e176. rhamnose. Biotechnology and Bioengineering, 33, 365e368.
Dagbert, C., Meylheuc, T., & Bellon-Fontaine, M. N. (2006). Corrosion Lukondeh, T., Ashbolh, N. J., & Rogers, P. L. (2003). Evaluation of
behavior of AISI 304 stainless steel in presence of a biosurfactant Kluyveromyces marxianus FII 510700 grown on a lactose-based
M. Nitschke, S.G.V.A.O. Costa / Trends in Food Science & Technology 18 (2007) 252e259 259

medium as a source of natural bioemulsifier. Journal of Industrial Poremba, K., Gunkel, W., Lang, S., & Wagner, F. (1991b). Toxicity
Microbiology & Biotechnology, 30, 715e720. testing of synthetic and biogenic surfactants on marine microor-
Makkar, R. S., & Cameotra, S. S. (1997). Utilization of molasses for ganisms. Environmental Toxicology and Water Quality, 6(2),
biosurfactant production by two Bacillus strains at thermophilic 157e163.
conditions. Journal of the American Oil Chemists’ Society, 74, Pruthi, V., & Cameotra, S. S. (1997). Production, properties of a
887e889. biosurfactant synthesized by Arthrobacter protophormiae e an
Makkar, R. S., & Cameotra, S. S. (2002). An update to the use of Antartic strain. World Journal of Microbiology & Biotechnology,
unconventional substrates for biosurfactant production and their 13, 137e139.
new applications. Applied Microbiology and Biotechnology, 58, Rodrigues, L., Banat, I. M., Teixeira, J., & Oliveira, R. (2006).
428e434. Biosurfactants: potential applications in medicine. Journal of
McInerney, M. J., Javaheri, M., & Nagle, D. P. (1990). Properties of the Antimicrobial and Chemotherapy, 57(4), 609e618.
biosurfactant produced by Bacillus liqueniformis strain JF-2. Rodrigues, L., Moldes, A., Teixeira, J., & Oliveira, R. (2006). Kinetic
Journal of Industrial Microbiology & Biotechnology, 5, study of fermentative biosurfactant production by Lactobacillus
95e102. strains. Biochemical Engineering Journal, 28, 109e116.
Mercade, M. E., & Manresa, M. A. (1994). The use of agroindustrial Rodrigues, L. R., Banat, I. M., van der Mei, H. C., Teixeira, J. A., &
by-products for biosurfactant production. Journal of the American Oliveira, R. (2006). Interference in adhesion of bacteria and yeasts
Oil Chemists’ Society, 71, 61e64. isolated from explanted voice prostheses to silicone rubber by
Meylheuc, T., Methivier, C., Renault, M., Herry, J. M., Pradier, C. M., & rhamnolipid biosurfactants. Journal of Applied Microbiology,
Bellon-Fontaine, M. N. (2006). Adsorption on stainless steel 100(3), 470e480.
surfaces of biosurfactants produced by gram-negative and Rosenberg, E., & Ron, E. Z. (1999). High- and low-molecular-mass
gram-positive bacteria: consequence on the bioadhesive behaviour microbial surfactants. Applied Microbiology and Biotechnology,
of Listeria monocytogenes. Colloids and Surfaces B: Biointerfaces, 52, 154e162.
52, 128e137. Shepherd, R., Rockey, J., Sutherland, I. W., & Roller, S. (1995). Novel
Meylheuc, T., Renault, M., & Bellon-Fontaine, M. N. (2006). bioemulsifiers from microorganisms for use in foods. Journal of
Adsorption of a biosurfactant on surfaces to enhance the Biotechnology, 40, 207e217.
disinfection of surfaces contaminated with Listeria monocytogenes. Shete, A. M., Wadhawa, G., Banat, I. M., & Chopade, B. A. (2006).
International Journal of Food Microbiology, 109, 71e78. Mapping of patents on bioemulsifier and biosurfactant: a
Meylheuc, T., van Oss, C. J., & Bellon-Fontaine, M. N. (2001). review. Journal of Scientific and Industrial Research, 65(2),
Adsorption of biosurfactant on solid surfaces and consequences 91e115.
regarding the bioadhesion of Listeria monocytogenes LO 28. Singh, P., & Cameotra, S. S. (2004). Potential applications of microbial
Journal of Applied Microbiology, 91, 822e832. surfactants in biomedical sciences. Trends in Biotechnology, 22(3),
Mireles II, J. P., Toguchi, A., & Harshey, R. M. (2001). Salmonella 142e146.
enterica serovar typhimurium swarming mutants with altered Syldatk, C., Lang, S., & Wagner, F. (1985). Chemical and physical
biofilm-forming abilities: surfactin inhibits biofilm formation. characterization of four interfacial-active rhamnolipids from
Journal of Bacteriology, 183(20), 5848e5854. Pseudomonas sp. DSM 2874 grown on alkanes. Zeitschrift fur
Mohan, P. K., Nakhla, G., & Yanful, E. K. (2006). Biokinetics of Naturforschung C, 40, 51e60.
biodegradability of surfactants under aerobic, anoxic and Thompson, D. N., Fox, S. L., & Bala, G. A. (2000). Biosurfactants from
anaerobic conditions. Water Research, 40, 533e540. potato process effluents. Applied Biochemistry and Biotechnology,
Mulligan, C. N. (2005). Environmental applications for biosurfactants. 84e86, 917e929.
Environmental Pollution, 133, 183e198. Van Haesendonck, I. P. H., & Vanzeveren, E. C. A. (2004). Rhamno-
Nitschke, M., Costa, S. G. V. A. O., & Contiero, J. (2005). Rhamnolipid lipids in bakery products. W.O. 2004/040984, International
surfactants: an update on the general aspects of these remarkable application patent (PCT).
biomolecules. Biotechnology Progress, 21(6), 1593e1600. Velikonja, J., & Kosaric, N. (1993). Biosurfactants in food applications.
Nitschke, M., Costa, S. G. V. A. O., Haddad, R., Gonçalves, L. A. G., In N. Kosaric (Ed.), Biosurfactants: Production, properties and
Eberlin, M. N., & Contiero, J. (2005). Oil wastes as unconven- applications (pp. 419e446). New York: Marcel Dekker.
tional substrates for rhamnolipid biosurfactant production by Velraeds, M. M. C., van der Mei, H. C., Reid, G., & Busscher, H. J.
Pseudomonas aeruginosa LBI. Biotechnology Progress, 21(5), (1996). Inhibition of initial adhesion of uropathogenic
1562e1566. Enterococcus faecalis by biosurfactants from Lactobacillus
Nitschke, M., & Pastore, G. M. (2003). Cassava flour wastewater as isolates. Applied and Environmental Microbiology, 62(6), 1958e
a substrate for biosurfactant production. Applied Biochemistry and 1963.
Biotechnology, 105e108, 295e301. Vollenbroich, D., Özel, M., Vater, J., Kamp, R. M., & Pauli, G.
Nitschke, M., & Pastore, G. M. (2004). Biosurfactant production by (1997). Mechanism of inactivation of enveloped viruses by the
Bacillus subtilis using cassava processing effluent. Applied biosurfactant surfactin from Bacillus subtilis. Biologicals, 25(3),
Biochemistry and Biotechnology, 112, 163e172. 289e297.
Nitschke, M., & Pastore, G. M. (2006). Production and properties of Wang, X., Gong, L., Liang, S., Han, X., Zhu, C., & Li, Y. (2005).
a surfactant obtained from Bacillus subtilis grown on cassava Algicidal activity of rhamnolipid biosurfactants produced by
wastewater. Bioresource Technology, 97, 336e341. Pseudomonas aeruginosa. Harmful Algae, 4, 433e443.
Patel, R. M., & Desai, A. J. (1997). Biosurfactant production by Yoo, D. S., Lee, B. S., & Kim, E. K. (2005). Characteristics of microbial
Pseudomonas aeruginosa GS3 from molasses. Letters in Applied biosurfactant as an antifungal agent against plant pathogenic
Microbiology, 25, 91e94. fungus. Journal of Microbiology & Biotechnology, 15(6), 1164e
Poremba, K., Gunkel, W., Lang, S., & Wagner, F. (1991a). Marine bio- 1169.
surfactants, III. Toxicity testing with marine microorganisms and Zottola, E. A. (1994). Microbial attachment and biofilm formation:
comparison with synthetic surfactants. Zeitschrift fur a new problem in the food industry? Food Technology, 48,
Naturforschung, 46, 210e216. 107e114.

You might also like