Percentages of growth inhibition obtained with the biosurfactant Rufisan isolated from
1. Introduction
Several compounds with tensioactive properties are synthesized by living organisms, from plants (e.g.
Physical and chemical properties, surface tension reduction, and stability of the emulsion formed are important characteristics in biosurfactant that make possible its use in countless biological applications. Most work on biosurfactant applications has been focused on their use in environmental applications owing to their diversity, environmentally friendly nature, suitability for large-scale production and selectivity [3]. Biosurfactants have several advantages over chemical surfactants, such as lower toxicity, higher biodegradability and effectiveness at extreme temperatures or pH values [4]. Many of the potential applications that have been considered for biosurfactants depend on whether they can be produced economically; however, much effort in process optimization and at the engineering and biological levels have been carried out [5]. Despite their potential and biological origin only a few studies have been carried out on applications related to the biomedical field [6]. Some biosurfactants are suitable alternatives to synthetic medicines and antimicrobial agents and may be used as safe and effective therapeutic agents [6].
Furthermore, biosurfactants have been found to inhibit the adhesion of pathogenic organisms to solid surfaces or to infection sites hampering biofilm formation that is the cause of many diseases, as for example cystic fibrosis [7]. Therefore, prior adhesion of biosurfactants to solid surfaces might constitute a new and effective means of combating colonization by pathogenic microorganisms and subsequent biofilm formation [8,9].
Pre-coating vinyl urethral catheters by running a surfactin solution through them before inoculation with media resulted in a decrease in the amount of biofilm formed by
Microbial surfactants are not yet competitive with those produced by the chemical industry, but efforts should be made on the different production aspects to find suitable and economic substrates and to develop new strategies to increase the volumetric productivity. We have shown that the co-utilization of ground-nut oil refinery residue and corn steep liquor is an attractive choice for biosurfactant production. The biosurfactant adhesive mechanism is based in the inhibition of microorganisms to different surfaces can interact with interfaces of the molecule. In this sense, they are an alternative to synthetic surface-active agents because of their low toxicity and biodegradability [7,12].
Considering the lack of studies with yeasts biosurfactants for medical purposes, and is an attractive characteristics showed by produced by the
2. Adhesion and microbial biofilms
The interest of biosurfactants as alternative medicines and antimicrobial agents increased considering the safe use as effective therapeutic agent on human and animal cells [12]. In consequence, the microbial adhesion is mediated by specific interactions between cells surface structures and molecular mass on the substratum surface, or by non-specific interaction forces, including electrostatic forces, acid-base interactions and Vander Waals forces [13].
during exponential growth, presumably as a result of increased cell wall hydrophobicity during this growth phase
The conditioning film on the biomaterial surface (and on the bacterial cell surface) plays an important role, as it changes the physicochemical properties of the interacting surfaces. Albumin is a strong adhesion inhibitor, for unknown reasons, although changes in hydrophobiciy and sterical hindrance are proposed as mechanisms [14].
The adhesion of microorganisms to a surface is one of the first stages in the development of a biofilm and is believed to be influenced by a number of factors. As the substrate is essential in the development of a biofilm, an understanding of how substrate properties affected the adherence of bacterial cells my assist in designing or modifying substrates inhibitory to bacterial adhesion. Many of these molecules are proteinaceous constitution, such as serum albumin, fibrogen and collagen, and some have been shown to affect subsequent bacterial adhesion [13].
The formation of infectious biofilm on biomaterial appeared to involve several sequential steps. Immediately after exposure of a device to body fluids, such as blood, saliva, or urine, macromolecular components adsorb to form a conditioning film [15]. The most microbial surfactants are complex molecules, comprising different structures that include peptides, glycolipids, glycopeptides, fatty acids and phospolipids, as reviewed recently. Among the many classes of biosurfactants, lipopeptides are particularly interesting because their high surface activities and antibiotic potential. Lipopeptides are molecules act as antibiotics, antiviral and antitumoral agents, and enzyme inhibitors. Those molecules enhance or decrease the bacterial surface hydrophobicity following that the surface is less or more hydrophobic[16]. Morikawa et al. [17] identified and characterized a biosurfactant, arthrofactin, produced by
Glycolipids are the most common class of biosurfactans of which the most effective from the point of view of surface active properties arethe trehalose lipids of
3. Antimicrobial activity of biosurfactant
Several biosurfactants which exhibit antimicrobial activity against various microorganisms have been previously described. They include surfactin and iturin produced by
Among the genus
The evaluation of the antimicrobial activity of these compounds was carried out against 29 bacteria.
The antimicrobial activity of the crude biosurfactant isolated from
The biosurfactant was effective against the microorganisms tested, albeit to different degrees. The highest anti-adhesive percentages were obtained for a biosurfactant concentration of 12 mg/l or 4xCMC. Non-pathogenic species associated with the oral cavity of
The growth of the other microorganisms tested was poorly inhibited. Percentages of 5%, 5%, 15%, 16% and 18% were observed for
The antimicrobial activity of the biosurfactant isolated from
The tested biosurfactant presented antimicrobial activity against all microorganisms used, although, depending on the microorganism, the biosurfactant presents different effective concentrations. The highest concentration of biosurfactant tested (10 mg ml-1) showed high percentages of inhibition for
The crude biosurfactant showed antimicrobial activity against a broad range of micro-organisms, including Gram-positive and Gram-negative bacteria and yeasts.
Biosurfactants antimicrobial activity has been described, as for example surfactin, a
Some biosurfactants are able, even in low concentrations, to destabilize the microorganism’s membranes, killing them or disabling their growth [29, 30]. The interest in biosurfactants was first expressed due to its potential antimicrobial properties, being the first reported and actually the most studied biosurfactants, rhamnolipid and surfactin [31]. Gram-positive bacteria are more sensitive to biosurfactants than Gram-negative bacteria, which are weakly inhibited or not inhibited at all [32].
Several biosurfactants that exhibit antimicrobial activity have been previously described. However, there are few reports about the antimicrobial activity of biosurfactants isolated from
4. Anti-adhesive activity of biosurfactant
Involvement of biosurfactants in microbial adhesion and desorption has been widely described, and adsorption of biosurfactants to solid surfaces might constitute an effective strategy to reduce microbial adhesion and combating colonization by pathogenic microorganisms, not only in the biomedical field, but also in other areas, such as the food industry [36, 37].
Biosurfactants have been found to inhibit the adhesion of pathogenic organisms to solid surfacesor to infection sites, thus prior adhesion of biosurfactant to solid surfaces might constitute a new and effective means of combating colonization by pathogenic microorganisms [12]. Precoating vinyl urethral catheters by running a surfactin solution through them before inoculation with media resulted in a decrease of the amount of biofilm formed by
In addition to the antimicrobial properties, the anti-adhesive activity of the biosurfactant was evaluated against a variety of bacterial and fungal strains. The biosurfactant showed anti-adhesive activity against most of the microorganisms tested, but the anti-adhesive effect depends on the concentration and the micro-organism tested (Table 3).
The crude biosurfactant showed anti-adhesive activity against most of the microorganisms tested from the minimum concentration used (0.75 mg/l). The anti-adhesive property was proportional to the concentration of the biosurfactant. For the microorganisms of the
Gudina et al. [24] observed an anti-adhesive activity for the biosurfactant from
The use and potential commercial applications of biosurfactants in the medical field has increased considerably in the last years. Their antimicrobial and anti-adhesive properties make them relevant molecules for use in combating many diseases and infections and as therapeutic agents [18].
Adhesion to surfaces and subsequent biofilm formation consist in a surviving strategy used by microorganisms in several hostile environments, protecting them from dehydration, predators, biocides and extreme conditions [22]. The antiadhesive activity of this biosurfactant was evaluated against a variety of bacterial and fungal strains. The biosurfactant showed antiadhesive activity against most of the microrganisms tested, but the antiadhesive effect depends on the concentration and the microrganism tested (Table 4).
This biosurfactant was effective against all the microorganisms tested, albeit to different degree. With regard to the
Regarding the yeast, a total inhibition of adhesion was also observed for
The antiadhesive activity of the crude biosurfactant isolated from
Besides possessing
The adsorption of a biosurfactant on surface was found to change its hydrophobicity, which might caused interference in the adhesion and desorption processes [41]. Furthermore, Velraeds et al. [42] reported the inhibition of adhesion of pathogenic enteric bacteria by a biosurfactant produced by
5. Conclusion
In conclusion, in this work we have demonstrated the antimicrobial and anti-adhesive properties of the crude biosurfactant isolated from
Acknowledgement
This work was financially supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE), Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Braga, Portugal. We are grateful to Núcleo de Pesquisas em Ciências Ambientais (NPCIAMB) laboratories, Universidade Católica de Pernambuco, Brazil.
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