Antimicrobial and anti-adhesive potential of a biosurfactant Rufisan produced by Candida lipolytica UCP 0988.

In the last years, researches developed with biosurfactants for application in the medical area have been revealing the promising biological activities of these biomolecules. In this work the antimicrobial and anti-adhesive properties of a biosurfactant Rufisan isolated from the yeast Candida lipolytica UCP 0988, growth in a medium supplemented with ground nut refinery residue was determined against several microorganisms. The biosurfactant was able to reduce the water surface tension from 70 to 25.3 mN/m and showed a critical micelle concentration (CMC) of 0.03%. The biosurfactant was isolated after 72 h of fermentation and was tested in concentrations varying from 0.75 to 12 mg/l. The highest antimicrobial activities were observed against Streptococcus agalactiae, Streptococcus mutans, Streptococcus mutans NS, Streptococcus mutans HG, Streptococcus sanguis 12, Streptococcus oralis J22 at a concentration superior to the biosurfactant critical micelle concentration. Moreover, the biosurfactant showed anti-adhesive activity against most of the microorganisms tested. As far as we know, this is the first compilation of data on antimicrobial and anti-adhesive activities of a biosurfactant obtained from a Candida strain against such a broad group of microorganisms. The results obtained in this work showed that the biosurfactant from C. lipolytica is a potential antimicrobial and/or anti-adhesive agent for several biomedical applications.

Optimizing the medium components in bioemulsifiers production by Candida lipolytica with response surface method.

A response surface methodology was used to study bioemulsifier production by Candida lipolytica. A 2(4) full experimental design was previously carried out to investigate the effects and interactions of the concentrations of corn oil, urea, ammonium sulfate, and potassium dihydrogen orthophosphate on the emulsification activity (EA) of the bioemulsifier produced by C. lipolytica. The best EA value (3.727 units of emulsification activity (UEA)) was obtained with a medium composed of 0.4 g of urea, 1.1 g of ammonium sulfate, 2.04 g of potassium dihydrogen orthophosphate, 5 mL of corn oil, 50 mL of distilled water, and 50 mL of seawater. A curvature check was performed and revealed a lack of fit of the linear approximation. The proximity of the optimum point was evident, as was the need for quadratic model and second-order designs that incorporate the effect of the curvature. Medium constituents were then optimized for the EA using a three-factor central composite design and response surface methodology. The second-order model showed statistical significance and predictive ability. It was found that the maximum EA produced was 4.415 UEA, and the optimum levels of urea, ammonium sulfate, and potassium dihydrogen orthophosphate were, respectively, 0.544% (m/v), 2.131% (m/v), and 2.628% (m/v).

Screening of chitin deacetylase from Mucoralean strains (Zygomycetes) and its relationship to cell growth rate.

Chitin deacetylase (CDA) is an enzyme that catalyzes the hydrolysis of acetamine groups of N-acetyl-D: -glucosamine in chitin, converting it to chitosan in fungal cell walls. In the present study, the activity in batch culture of CDA from six Mucoralean strains, two of them wild type, isolated from dung of herbivores of Northeast Brazil, was screened. Among the strains tested, Cunninghamella bertholletiae IFM 46114 showed a high intracellular enzyme activity of 0.075 U/mg protein after 5 days of culture, and a wild-type strain of Mucor circinelloides showed a high intracellular enzyme activity of 0.060 U/mg protein, with only 2 days of culture, using N-acetylchitopentaose as substrate. This enzyme showed optimal activity at pH 4.5 in 25 mM glutamate-sodium buffer at 50 degrees C, and was stable over 1 h preincubation at the same temperature. The kinetic parameters of CDA did not follow Michaelis-Menten kinetics, but rather Hill affinity distribution, showing probable allosteric behavior. The apparent K(HILL) and Vmax of CDA were 288+/-34 nmol/l and 0.08+/-0.01 U mg protein(-1) min(-1), respectively, using N-acetylchitopentaose as substrate at pH 4.5 at 50 degrees C.

Decolorization of reactive azo dyes by Cunninghamella elegans UCP 542 under co-metabolic conditions.

The inappropriate disposal of dyes in wastewater constitutes an environmental problem and can cause damage to the ecosystem. Alternative treatments have been reported that fungi are particularly effective in the decolorization of textile effluents. The decolorization of dyes with different molecular structures by Cunninghamella elegans was evaluated under several media conditions. The decolorization procedures consisted of adding 72 h of mycelium into the culture medium containing either orange or reactive black or reactive red or a mixture of these dyes in the presence or absence of sucrose and/or peptone. The decolorization profile was highly dependent upon the incubation time, the molecular structure of the dye and presence or absence of co-substrates. The presence of sucrose or both sucrose and peptone significantly increased the decolorization of the solutions, however, the presence of only the nitrogen source suppressed it. The ultraviolet spectra of the solutions before and after decolorization suggested the occurrence of biodegradation in addition to the biosorption of the dyes. All tested dyes, except for the reactive black, caused inhibition of respiration of Escherichia coli, which suggested that toxic metabolites were produced.

Chitosan from Syncephalastrum racemosum used as a film support for lipase immobilization.

Chitosan from a native Mucoralean strain, Syncephalastrum racemosum, isolated from herbivorous dung (Northeast-Brazil), was used as a film support for lipase immobilization. S. racemosum showed highest chitosan yield (152 mg g dry mycelia weight(-1); 15.2% of dry mycelia weight) among the nine strains screened, which presented 89% D-glucosamine. A chitosan film was used for lipase (EC 3.1.1.3) immobilization using glutaraldehyde as a bifunctional agent. The immobilized lipase retained 47% (12.6 micromol s(-1) m(-2)) of its initial catalytic activity after four cycles of reaction. This result is comparable (same order of magnitude) to that of the enzyme immobilized on film made from commercially available crustacean chitosan.

Typing of human and bovine Staphylococcus aureus by RAPD-PCR and ribotyping-PCR.

AIMS: To investigate genetic diversity among Staphylococcus aureus and to delineate the geographical distribution of the strains found. METHODS AND RESULTS: RAPD-PCR and ribotyping-PCR were employed for the characterization of Staph. aureus isolates from bovine and nosocomial origin. Among the strains, five to nine groups were distinguished by RAPD-PCR, depending on which primer was used, while ribotyping-PCR distinguished seven ribotypes. CONCLUSIONS, AND SIGNIFICANCE AND IMPACT OF THE STUDY: These results demonstrate the genetic heterogeneity of the strains studied, and the large dissemination of some clones throughout different regions and hosts, findings that may allow the monitoring of Staph. aureus infections.

Physical and rheological characterisation of polyethylene glycol-cashew-nut tree gum aqueous two-phase systems.

The characterisation of the polyethylene glycol-cashew-nut tree gum aqueous two-phase system is described. Factors which affect the phase diagram including polymer molecular mass, pH and temperature were analysed. The physico-chemical properties of the system such as density, viscosity, volume ratio and phase separation times were also described. The characteristics of the system studied indicate it to be very attractive as a separation technique.

Bioemulsifier production in batch culture using glucose as carbon source by Candida lipolytica.

The yeast Candida lipolytica IA 1055 produced an inducible extracellular emulsification activity while utilizing glucose at different concentrations as carbon source during batch fermentation at 27 degrees C. In all glucose concentrations studied, maximum production of emulsification activity was detected in the stationary phase of growth, after pH reached minimal values. The bioemulsifier isolated was a complex biopolymer constituting proteins, carbohydrates, and lipids. The results obtained in this work show that the biosynthesis of a bioemulsifier is not simply a prerequisite for the degradation of extracellular hydrocarbon.

Biochemical markers in taxonomy of the genus Cunninghamella.

The chemical composition of fatty acids and ubiquinones was studied in 18 strains of Cunninghamella, to establish quantitative and qualitative differences within the genus. Fatty acids analysis has shown the presence of four groups. Ubiquinone analysis, through high performance liquid chromatography (HPLC), demonstrated the existence of three different groups based on the ubiquinone type. The average percentage of fatty acids of the species Cunninghamella elegans and Cunninghamella bertholletiae, show variations in linolenic and stearic acids, suggesting the possibility of differentiation between the two species.

A factorial design analysis of chitin production by Cunninghamella elegans.

Chitin production by Cunninghamella elegans (IFM 46109) was studied with a two-level full factorial design, varying time of cultivation and the concentration of D-glucose, L-asparagine, and thiamine in the culture medium. The material extracted was characterized by infrared and NMR spectroscopy. The highest chitin yield, 28.8%, was comparable with the highest in the literature and was obtained with a medium containing 60 g.L-1 of glucose, 3 g.L-1 of asparagine, and 0.008 mg.L-1 of thiamine. Increasing the time of cultivation from 24 h to 72 h did not affect chitin production. The three factors showed significant positive effects on chitin production, without interactions between them.

New aqueous two-phase system based on cashew-nut tree gum and poly(ethylene glycol).

The characterisation of a new system based on cashew-nut tree gum, a branched acidic heteropolysaccharide found in Brazil, and poly(ethylene glycol) (PEG) was studied. Phase diagrams are provided for the PEG-cashew-nut tree gum system. The influence of PEG molecular mass, tie-line length and pH on bovine serum albumin (BSA) partition was investigated. Protein partition coefficient was little influenced by changing PEG molecular mass. Increasing the tie-line length decreased the partition. Increasing the pH also raised the BSA partition coefficient. It is shown that systems formed by PEG and cashew-nut tree gum may be considered as an interesting alternative for use in protein purification.

The use of babassu oil as substrate to produce bioemulsifiers by Candida lipolytica.

Candida lipolytica IA 1055 produced an extracellular emulsifier when using babassu oil as its sole carbon source during batch and fed batch fermentations at 27 degrees C. Emulsification activity was detected after 60 h of growth in all conditions studied. The bioemulsifier was isolated after 144 h of fermentation from the best condition studied. The biopolymer seems to be a polysaccharide-protein-lipid complex.

Actin cytoskeleton structure and its localization in gongronella butleri, mucor javanicus and absidia cylindrospora (mucorales-zygomycetes)

Mediante estudios de microscopia òptica de fluorescencia se describe y caracteriza la estructura, distribución y localización de la actina, en tres especies de hongos de la clase zygomycetes: absidia cylindrospora, gongronella butleri y mucor javanicus. La estructura de la actina fué analizada utilizándose el marcador fitc-phalloidina, especifico para actina, en muestras fijadas con formaldehido. En los resultados se observan diferencias en la estructura, distribución y localización de esta proteina en las especies analizadas. Filamentos , aglomerados, placas o aglomerados periféricos y gránulos, fueron las formas más comunes encontradas en los hongos, con prevalencia de los aglomerados observados en todos los citoplasma de las hifas. La tinción de actina en losápices de las hifas, solo se observó en mucor javanicus, esta diferencia en la estructura de la actina podría estar asociada a zonas de expansión de la pared celular de los hongos

Effects of tween 80 on protease production by candida lipolytica

Se estudió la acción del tween 80, agente tensoactivo no iónico, sobre la proteasa producida por candida lipolytica. El microorganismo fue cultivado en medio de cultivo conteniendo diferentes concentraciones del agente, el cual fue introducido en el medio en diferentes intervalos de tiempo. La presencia del agente en el medio de cultivo incrementó la actividad de la proteasa extracelular e intracelular y el crecimiento del hongo. Los efectos del agente podrían estar relacionados con el tiempo de adición y las concentraciones empleadas en el medio de cultivo

Estudo comparativo da produçäo de bioemulsificantes por amostras de Candida lipolytica / Comparative study on bioemulsifiers produced by Candida lipolytica strains

This study was carried out to evaluate the production of bioemulsifiers by two strains of Candida lipolytica 1055 and 1120 respectively. The results obtained showed that strain 1055 was the best producer in media supplemented with 5 per cent of Babaçu oil and glucose 1 per cent, used as carbon source. By using these carbon sources it was possible to show that the bioemulsifiers were produced by Candida lipolytica in the end of exponential growth phase and begining of stationary growth phase as secondaries metabilites

Ubiquinone systems of the genus Cladosporium and morphologically similar taxa.

The ubiquinone (coenzyme Q) systems of 14 species of Cladosporium were determined. The genus was divided into two groups based on the distribution of the major ubiquinones, Q-10 and Q-10(H2). The group containing Q-10 consisted of six species, four of which were human pathogens, whereas the group containing Q-10(H2) consisted of eight plant pathogenic and/or saprophytic species. The results presented here agree with phylogenetic and physiological studies which have shown that the human-pathogenic species of Cladosporium represent a homogeneous, cohesive group.

Effect of sodium dodecyl sulfate on lipase of Candida lipolytica.

The effects of sodium dodecyl sulfate on extracellular lipase produced by Candida lipolytica have been studied. The microorganism was grown in culture medium containing different sodium dodecyl sulfate concentrations added to the culture at different intervals of growth. The extracellular lipase activity was not detected when the treated culture supernatants were directly tested in Yeast Mold Agar-Triolein-Rhodamine plates, regardless of surfactant addition time and concentrations. However, after ammonium sulfate precipitation and dialysis, the extracellular lipase activity could be recovered. Therefore, the surfactant, under the experimental conditions used here, does not seem to be able to inhibit lipase production, but it does inhibit the enzyme activity because of its presence in the mixture of the reaction.

Report of chronic subcutaneous abscesses caused by Exophiala spinifera.

A case of phaeohyphomycosis with subcutaneous abscesses involving skin, muscle and bone was caused by Exophiala spinifera. This is the first report of E. spinifera causing bone degeneration in Recife-PE, Brazil.

Bacitracin production by a new strain of Bacillus subtilis. Extraction, purification, and characterization.

A new strain of Bacillus subtilis C 126 was isolated from sugar cane fermentation and produced an antibiotic that inhibited the growth of Micrococcus flavus. The production of the antibiotic in culture medium followed to extraction with n-butanol, thin layer chromatography, and microbiological tests indicated that a polypeptide antibiotic was produced. The fraction obtained by Sephadex G-25 column and analyzed by HPLC indicated that bacitracin complex was produced.