Sandra Aparecida de Assis

Aqueous extraction of pectin from sisal waste

In this work, sisal waste was used as a source of pectin. Sisal is known worldwide as a source of hard fibres, and Brazil is the largest producer of sisal, producing more than 246,000 tonnes. However, the process of removing the fibres of the sisal leaf generates 95% waste. This study investigated the effect of the liquid/solid ratio (%), time (min), and temperature (°C) on the yield of the pectin obtained from sisal waste by attractive environmentally friendly process. A statistical Box-Behnken design was applied to determine the important effects and interactions of these independent variables on the yield of pectin, the dependent variable. Significant models were obtained. The yield of the extracted pectin ranged from 4.61 to 19.2%. The conditions that produced the highest yield (19.2%) were a temperature of 85 °C, extraction time of 60 min and a liquid/solid ratio of 2%.

Antioxidant activity, ascorbic acid and total phenol of exotic fruits occurring in Brazil.

The antioxidant activity, ascorbic acid and phenolic content were studied in 10 exotic fruits from Brazil: abiu, acerola, wax jambu, cashew, mamey sapote, carambola or star fruit, Surinam cherry, longan, sapodilla and jaboticaba. The ascorbic acid was determined by 2,6-dichloroindophenol titrimetic methods and total phenols were measured colorimetrically using the Folin–Ciocalteu reagent. The antioxidant activity was investigated with three different methods: hypochlorous acid scavenging activity, 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization assay, and 2,2-diphenyl-1-picrylhydrazyl radical scavenging method. The highest content of vitamin C (1,525.00 mg/100 g pulp) occurred in acerola. The total phenol content was higher in abiu, acerola, Surinam cherry and sapodilla. In relation to antioxidant activity, acerola has showed the great values in all three different methods tested. It was found that the fruits have a significant antioxidant effect when tested by each method, respectively, and these antioxidant capacities are promising. The sample concentration also influenced its antioxidant power.

Production, extraction and characterization of exopolysaccharides produced by the native Leuconostoc pseudomesenteroides R2 strain

The genus Leuconostoc belongs to a group of lactic acid bacteria usually isolated from fermented vegetables, which includes species involved in the production of exopolysaccharides (EPS). These biopolymers possess considerable commercial potential. Because of the wide variety of industrial applications of EPS, this study aimed to produce and characterize the native exopolysaccharide strain Leuconostoc pseudomesenteroides R2, which was isolated from cabbage collected in a semi-arid region of Bahia. We employed the following conditions for the production of EPS: 10.7% sucrose, pH 8.2, without agitation and incubation at 28ºC for 30 hours. The fermentation broth was treated with ethanol and generated two types of polysaccharide substances (EPS I and EPS II). The identification of EPS I and EPS II was conducted using FT-IR, (1)H, (13)C and DEPT-135 NMR spectra. The two substances were identified as linear dextran α polysaccharides (1 → 6) which indicated different characteristics with respect to thermal analysis and density of free packaging, viscosity and time of solubilization. Both dextrans are of low density, possess high thermal stability and exhibited the behavior characteristic of pseudoplastic polymers.

Synthesis of naringin 6"-ricinoleate using immobilized lipase

BackgroundNaringin is an important flavanone with several biological activities, including antioxidant action. However, this compound shows low solubility in lipophilic preparations, such as is used in the cosmetic and food industries. One way to solve this problem is to add fatty acids to the flavonoid sugar unit using immobilized lipase. However, there is limited research regarding hydroxylation of unsaturated fatty acids as an answer to the low solubility challenge. In this work, we describe the reaction of naringin with castor oil containing ricinoleic acid, castor oils major fatty acid component, using immobilized lipase from Candida antarctica. Analysis of the 1H and 13 C NMR (1D and 2D) spectra and literature comparison were used to characterise the obtained acyl derivative.ResultsAfter allowing the reaction to continue for 120 hours (in acetone media, 50°C), the major product obtained was naringin 6″-ricinoleate. In this reaction, either castor oil or pure ricinoleic acid was used as the acylating agent, providing a 33% or 24% yield, respectively. The chemical structure of naringin 6″-ricinoleate was determined using NMR analysis, including bidimensional (2D) experiments.ConclusionUsing immobilized lipase from C. antarctica, the best conversion reaction was observed using castor oil containing ricinoleic acid as the acylating agent rather than an isolated fatty acid.Graphical abstract

An alternative method for screening lactic acid bacteria for the production of exopolysaccharides with rapid confirmation

The accumulation of exopolysaccharides (EPS) produced by microorganisms occurs in the presence of excess substrate and limiting conditions of elements that are essential to growth, such as nitrogen, phosphorus, sulfur, and magnesium. The presence of EPS produced by bacterial cells contributes to slime colonies formation in solid medium and increased viscosity in liquid medium. This paper proposes an alternative method for screening EPS-producing lactic acid bacteria using solid medium-containing discs of filter paper that are saturated with active cultures. The screening was carried out under different culture conditions varying the type of sugar, pH, and temperature. EPS production was visualized by the presence of mucoid colonies on the discs, which was confirmed by the formation of a precipitate when part of this colony was mixed with absolute alcohol. The established conditions for obtaining a high number of isolates producing EPS were 10% sucrose, pH 7.5 and 28 oC. This method proved to be effective and economical because several strains could be tested on the same plate, with immediate confirmation.

Thermostable inulinases secreted by yeast and yeast-like strains from the Brazilian semi-arid region

The use of inulinases provides an alternative to the chemical process of inulin hydrolysis to obtain fructose syrup, and can reduce processing steps, time, and costs in the food industry. The objective of this work was to screen the thermostable inulinases produced by yeast and yeast-like strains isolated from the Brazilian semi-arid region. Thermostability was studied at different temperatures (60°C, 70°C, 80°C and 90°C) and for increasing periods of time (0–50 min). Thirty-three microorganisms were tested, and 27 showed inulinase activity with specific activities ranging from 0.98 to 73.79 µmol/mg protein/min. Three strains (CCMB 300, CCMB327 and CCMB328) showed the desired combination of high specific activity and a small reduction in residual activity when submitted to heat treatment (≥60°C). Our results indicate that the inulinases produced by these three yeast strains from the Brazilian semi-arid region have great potential to be used for inulin hydrolysis in the food industry.

Extraction optimization and antinociceptive activity of (1→3)-β-d-glucan from Rhodotorulamucilaginosa.

β-d-glucans are polymers of d-glucose monomers found in the cell walls of many bacteria, plants, fungi and yeasts. A variety of β-d-glucans differing in structures have been isolated from various sources and their biological activity to be regulated by various structural factors, such as the primary structure, molecular weight, solubility, and conformation. This study investigated the effect of extraction time and temperature on the yield of β-d-glucan produced by Rhodotorulamucilaginosa. A statistical Doehlert design was applied to determine the important effects and interactions of these independent variables on the yield of β-d-glucan, the dependent variable. Significant models were obtained. The best yield was of 25% obtained after 128min of extraction in a temperature of 72°C. The polysaccharides were characterized as (1⟶3)-β-d-glucan by methods spectroscopic (FT-IR, (1)HNMR and (13)CNMR). In addition, the antinociceptive effect was evaluated using different experimental tests (acetic acid-induced writhing test, formalin test and tail immersion test). The (1⟶3)-β-d-glucan showed a potent peripheral antinociceptive effect, possibly by the inhibition of inflammatory mediators.

Production, purification and characterization of a thermostable β-1,3-glucanase (laminarinase) produced by Moniliophthora perniciosa

The enzyme glucanase from Moniliophthora perniciosa was produced in liquid medium and purified from the culture supernatant. A multivariate statistical approach (Response Surface Methodology - RSM) was employed to evaluate the effect of variables, including inducer (yeast extract) and fermentation time, on secreted glucanase activities M. perniciosa detected in the culture medium. The crude enzyme present in the supernatant was purified in two steps: precipitation with ammonium sulfate (70%) and gel filtration chromatography on Sephacryl S-200. The best inducer and fermentation time for glucanase activities were 5.9 g L(-1) and 13 days, respectively. The results revealed three different isoforms (GLUI, GLUII and GLUIII) with purification factors of 4.33, 1.86 and 3.03, respectively. The partially purified enzymatic extract showed an optimum pH of 5.0 and an optimum temperature of 40°C. The enzymatic activity increased in the presence of KCl at all concentrations studied. The glucanase activity was highest in the presence of 0.2 M NaCl. The enzyme showed high thermal stability, losing only 10.20% of its specific activity after 40 minutes of incubation at 90°C. A purified enzyme with relatively good thermostability that is stable at low pH might be used in future industrial applications.

Polygalacturonase secreted by yeasts from Brazilian semi-arid environments

Microbial pectinolytic enzymes are known to play a commercially important role in a number of industrial processes. The objective of this study was to investigate the extracellular polygalacturonases of yeasts isolated from Brazilian semi-arid environments. Among the 250 colonies tested, only 33 produced extracellular polygalacturonases: Aureobasidium pullulans (18 isolates), Candida boidinii (one isolate), Trichosporonoides sp. (three isolates), Kluyveromyces marxianus (one isolate), Cryptococcus liquefaciens (one isolate), Pseudozyma sp. (four isolates), and yeast-like related to fungal endophyte (five isolates). The highest activity of polygalacturonase was observed in Pseudozyma sp. CCMB 300 (14.17±0.08 µmol acid galacturonic released/min/mg protein). This study shows the potential of yeasts and yeast-like organisms isolated from Brazilian semi-arid environments to produce pectinolytic enzymes.

Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis

This study aimed to verify the influence of pH and temperature on the lysis of yeast using experimental design. In this study, the enzymatic extract containing β-1,3-glucanase and chitinase, obtained from the micro-organism Moniliophthora perniciosa, was used. The experiment showed that the best conditions for lysis of Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) by lytic enzyme were pH 4.9 at 37 oC and pH 3.9 at 26.7 oC, respectively. The lytic enzyme may be used for obtaining various biotechnology products from yeast.