Bioaccessibility Evaluation of Soymilk Isoflavones with Biotransformation Processing.

Soy isoflavones are considered important sources of bioactive compounds, but they are poorly absorbable, due to their large hydrophilic structures. Some biotransformation strategies have been used to convert the glycosidic form into aglycones, making them available for absorption. This study evaluated the potential of enzymatic and/or microbial fermentation combined bioprocesses in a soymilk extract before and after gastrointestinal in vitro digestion. Commercial ß-glucosidase (ET) and a mix of commercial probiotics (F) containing Lactobacillus acidophilus, Lactobacillus casei, Lactococcus lactis, Bifidobacterium bifidum, and Bifidobacterium lactis were used to biotransform the soymilk phenolic extract. An isoflavone profile was identified using HPLC-DAD, total phenolic content was identified using the Folin-Ciocalteu test, and antioxidant capacity was identified using ORAC and FRAP. Soymilk enzymatically treated (ET) followed by microbial fermentation (ET + T) resulted in better conversion of glycosylated isoflavones (6-fold lower than control for daidzin and 2-fold for genistin) to aglycones (18-fold greater than control for dadzein and genistein). The total phenolic content was increased (3.48 mg/mL for control and 4.48 mg/mL for ET + T) and the antioxidant capacity was improved with treatments of ET + T (120 mg/mL for control and 151 mg/mL with ORAC) and with FRAP (285 µL/mL for control and 317 µL/mL). After the in vitro digestion, ET + T samples resulted in a higher content of genistein (two-fold higher than control); also, increases in the total phenolic content (2.81 mg/mL for control and 4.03 mg/mL for ET + T) and antioxidant capacity measured with ORAC were greater compared to undigested samples. In addition, the isolated microbial fermentation process also resulted in positive effects, but the combination of both treatments presented a synergistic effect on soy-based products.

Conditions of enzyme-assisted extraction to increase the recovery of flavanone aglycones from pectin waste.

The citrus pectin by-product (CPB), generated from pectin industry, is a rich-source of flavanones, but not explored until now. As most of these compounds are inside vacuoles or bound to cell wall matrix, enzymatic hydrolysis was applied on their recovery, followed by hydroalcoholic and ultrasound extraction. Different parameters were studied: enzymes (ß-glucosidase, tannase, and cellulase), their concentration (5, 10, and 20 U g-1 CPB), and reaction time (6, 12, and 24 h). Extracts were characterized in total phenolic content (TPC), antioxidant capacity (ORAC and DPPH assays), and polyphenolic profile (HPLC-DAD). All enzymatic treatments significantly improved CPB antioxidant capacity and TPC, compared with hydroalcoholic and ultrasound extraction. ß-glucosidase (5 U) for 24 h was the most effective in polyphenol extraction and bioconversion, followed by ß-glucosidase (5 U) for 12 h and tannase (5 U) for 24 h. Thus, the concentration of these enzymes was increased (10 and 20 U) to improve flavanones extraction. ß-glucosidase at 20 U offered the highest amount of naringenin (77.63 mg 100 g-1 of CPB) and hesperetin (766.44 mg 100 g-1) obtained so far by biological processes. According to Person's correlation analysis, TPC and antioxidant activity were highly correlated with CPB contents of hesperetin and naringenin. The aglycone flavanones are rarely found in natural sources and have higher biological potential than their glycosylated forms. Our results indicated enzyme-assisted extraction as a good choice for recovering aglycone flavanones from CPB, and increased knowledge on the biological activity of this agroindustrial waste, amplifying their application in food and pharmaceutical field.

Evaluation of Nanostructured Lipid Carriers Produced with Interesterified Buriti Oil.

RESEARCH BACKGROUND: Extracted from the pulp of an Amazonian fruit, buriti oil is rich in micronutrients with antioxidant properties and high biological value. The few studies available indicate that this oil could be used in a wide range of applications; however, there are no studies that work on the improvement in the characteristics of this oil for commercial application. The enzymatic interesterification is one of the tools available to improve the properties of oils and fats and our recent studies have demonstrated that the lipase could specifically act on buriti oil to produce structured lipids rich in oleic acid, while preserving most of the minor compounds present in this oil. Still looking for ways to expand the applicability of this raw oil, in this work, we are interested in studying the behaviour of this structured oil in nanostructured lipid carriers (NLCs). EXPERIMENTAL APPROACH: The NLCs were produced with interesterified buriti oil and the stability, droplet size, electrical charge, microstructure, polymorphism and antioxidant activity of the samples were evaluated by ORAC and FRAP methods. RESULTS AND CONCLUSIONS: The results showed that the interesterification formed more unsaturated triacylglycerols (TAGs), and NLCs prepared with interesterified buriti oil had smaller droplets than NLCs with crude buriti oil. Particles remained stable throughout the storage period and NLCs exhibited complex polymorphism with the presence of three crystalline forms. The oxygen radical absorbance capacity (ORAC) value was approx. 23% higher in nanolipid carries with structured lipids than in the nanolipid carriers with crude buriti oil, and the ferric reducing antioxidant power (FRAP) value 16% higher, demonstrating the influence of interesterification on the antioxidant activity of nanocarriers. Thus, NLCs prepared with interesterified buriti oil had small droplets, high stability and antioxidant capacity, and have a potential for nutritional and biological applications. NOVELTY AND SCIENTIFIC CONTRIBUTION: This research showed that interesterification positively influenced the physicochemical properties of NLCs, producing the oil rich in oleic acid, high stability and antioxidant capacity. Therefore, it may be interesting to use these nanocarriers to obtain efficient carrier systems for future applications.

Combined isoflavones biotransformation increases the bioactive and antioxidant capacity of soymilk.

Isoflavones are phenolic secondary metabolites mainly occurring in soy and soybean products. Compared to glycoside forms, isoflavone aglycones present higher biological activities. This study evaluated the potential of microbial and enzymatic treatments in biotransformed isoflavones in their biologically active forms in soymilk. Seven different cultures of lactic acid bacteria and bifidobacteria associated with the action of immobilized tannase enzyme were screened for isoflavone glycoside biotransformation ability. The biotransformed soymilk samples were characterized regarding isoflavone profile, total phenolic content, and in vitro antioxidant activities. All bacterial strains showed a good growth capacity in soymilk matrix and produced ß-glucosidase enzyme, which hydrolyzed isoflavone glycosides into aglycones in soymilk after 24 h of fermentation. The microbial fermentation followed by tannase reaction (FT processes) resulted in the highest increase of bioactive aglycones (10.3- to 13.1-fold for daidzein, 10.4- to 12.3-fold for genistein, and 3.8- to 4.7-fold for glycitein), compared to control soymilk. Further, FT processes enhanced the total phenolic content (53-70%) and antioxidant activity by ORAC (69-102%) and FRAP (49-71%) assays of the soymilk matrix. Therefore, the combination of microbial fermentation and tannase treatment is a promising strategy to obtain a fermented soy product rich in bioactive isoflavones with greater health-promoting potential. KEY POINTS: • Bacterial cultures and tannase enzyme displayed isoflavone deglycosylation activity. • The addition of tannase following the fermentation maximized the isoflavone conversion. • Increased isoflavone aglycones contributed to the improved antioxidant activity of soymilk.

Peanut skin polyphenols inhibit toxicity induced by advanced glycation end-products in RAW264.7 macrophages.

This is the first work to use a polyphenolic fraction derived from peanut skin to attenuate the toxicity induced by advanced glycation-end products (AGEs) in RAW264.7 macrophages. The RAW264.7 cells were stimulated by AGEs using the bovine serum albumin-fructose (BSA-FRU), bovine serum albumin-methylglyoxal (BSA-MGO) and arginine-methylglyoxal (ARG-MGO) models. The AGEs increased considerably the levels of reactive oxygen species and the gene expression of proinflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and nitric oxide. Twenty-eight polyphenols, including catechin, phenolic acids, and resveratrol were annotated in peanut skin extract (PSE) with the use of ultra-performance liquid chromatography coupled to quadrupole time of flight mass spectrometry (UPLC-QTOF/MSE) and to the UNIFI Scientific Information System. The administration of PSE at 100 and 150 µg/mL significantly inhibited oxidative stress, by suppressing the production of reactive oxygen species up to 70% and reducing the production of nitric oxide, IL-6 and TNF-α up to 1.7-, 10- and 107-fold, respectively.

Evaluation of cytotoxicity of nanolipid carriers with structured Buriti oil in the Caco-2 and HepG2 cell lines.

Buriti oil is rich in monounsaturated fatty acids, carotenoids and tocopherols and it is used for the treatment of various diseases. One strategy to restructure the triglycerides is enzymatic interesterification and nanocarriers have been employed to improve the solubility, bioavailability and stability of active compounds. This work aims to investigate the in vitro cytotoxicity of this structured oil in nanoemulsions and nanostructured lipid carriers to expand the applicability of the crude oil. None of the samples had a cytotoxic effect on Caco-2 and HepG2 cell lines at the concentrations tested. Structured lipids acted protecting against oxidative stress and lipid peroxidation. Additionally, no consumption of glutathione has been observed in both cells, and the compounds present in buriti oil are possibly acting as antioxidants. Thus, nanoparticles prepared with interesterified buriti oil had low cytotoxicity and high oxidative stability, with great potential for future applications.

Partial purification and biochemical characterization of an alkaline esterase from Sorghumbicolor

Esterases are enzymes that present good potential for industrial applications since they catalyze the formation or cleavage of ester bonds in water-soluble substrates, and sorghumseeds can represent an alternative source of this enzyme. The extraction of esterase from sorghumseeds is an economical alternative to obtain an enzyme of great interest. Esterases may improve the quality or accelerate the maturation of cheeses, cured bacon and fermented sausages and may also resolve racemic mixtures. Recently, seed esterases have been the focus of much attention as biocatalysts. In some cases, these enzymes present advantages over animal and microbial lipases due to some quite interesting features such as specificity and low cost, being a great alternative for their commercial exploitation as industrial enzymes The esterase studied here was extracted from sorghumseeds and some of its biochemical properties determined using synthetic substrates (p-nitrophenyl butyrate, caprylate, laurate and palmitate). The enzyme presented optimum activity at pH 8.0 and was stable in all the pH ranges studied. The optimum temperature for its activity was 40ºC but it showed low stability at this temperature (40% relative activity). The values derived for Km and Vmax were 0.67mM and 125 U.mg-1, respectively, obtained using p-nitrophenyl butyrate as the substrate. The enzyme showed an increase in activity when K2HPO4was added to the reaction medium, but the ions Mn2+, CO+, Hg+and Fe2+strongly inhibited the enzyme activity. This enzyme showed a preference for the hydrolysis of short chain fatty acids. The characteristics of sorghumesterase are very similar to those of the microbial esterases used in detergent processing.

Exploring in vitro effects of biotransformed isoflavones extracts: Antioxidant, antiinflammatory, and antilipogenic.

The present study aimed to investigate, in in vitro assays, the antilipogenic and antiinflammatory potential as well as the antioxidant capacity of biotransformed soymilk by tannase and ß-glycosidase enzymes. The results showed a significant enhancement of the antioxidant capacity, especially by biotransformed soymilk with free tannase (SFT), corresponding to an increase of 2.3 and 1.25 times by oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays, respectively. The lipid accumulation reduction by 3T3-L1 adipocytes assay was not significant. However, the antiinflammatory responses were expressive. In lipopolysaccharide-stimulated RAW 264.7 macrophages, SFT reduced around 37 times TNF-α expression at the highest tested concentration of the sample. Other inflammatory parameters, as IL-6 and nitric oxide, were no longer detected when the cells were treated with SFT and soymilk with immobilized enzymes, respectively. The biotransformed soy extracts with tannase have great potential to act as a nutraceutical, protecting the cells against oxidative damage and helping maintain health under inflammatory stress. PRACTICAL APPLICATIONS: Soy isoflavones have been associated with several beneficial effects on human health, including inhibition capacity of lipid accumulation in adipocytes, antiinflammatory properties, and antioxidant potential. However, the isoflavones bioavailability differs among their chemical forms, and studies have shown that the higher health benefits are conferred by aglycones and their metabolites, such as equol, compared to the other forms. For this reason, the enrichment of isoflavone aglycones and metabolites in soy-based products has attracted growing attention. The present study was focused on developing a bioprocess able to produce a rich extract with soy isoflavones metabolites, with increased bioactive potential for application as a functional ingredient or a nutraceutical.

Comparison of different Brazilian citrus by-products as source of natural antioxidants.

Significant amounts of citrus by-products remain after juice processing, which is then used to obtain pectin. The pectin industry then generates a new waste. No study has characterized this residue or suggested applications for it. The main goal of this study was to compare citrus industrial by-products that remain after juice (CJB) and pectin (CPB) extraction, aiming to obtain bioactive compounds. The residues were evaluated for their chemical composition, antioxidant capacity, and polyphenols content. CJB had 2-fold higher total phenols than CPB. Moreover, CJB exhibited higher antioxidant capacity than CPB. Nine polyphenols were detected; hesperidin was the main compound on both residues. CPB had higher content of polyphenols than CJB, which can be attributed to the industry procedure of pectin extraction. Thus, this study provides support for the reuse of CPB to obtain nutraceutical compounds, converting waste into added-value products.

Influence of rye flour enzymatic biotransformation on the antioxidant capacity and transepithelial transport of phenolic acids.

Phenolic acids have been reported to play a role on the antioxidant activity and other important biological activities. However, as most polyphenolics in food products are either bound to cellular matrices or present as free polymeric forms, the way they are absorbed has not been totally clear until now. Hydrolytic enzymes may act to increase functionalities in polyphenolic-rich foods, enhancing the bioaccessibility of phenolic compounds and minerals from whole grains. The aim of this study was to evaluate the action of tannin acyl hydrolase (tannase) on the total phenols, phenolic acid profile, antioxidant capacity and in vitro bioaccessibility of phenolic acids found in whole rye flour (RF). Besides increasing total phenols and the antioxidant capacity, tannase treatment increased the amounts of ferulic, sinapic and vanillic acids identified in RF, evidencing a new type of feruloyl esterase catalytic action of tannase. Vanillic and sinapic acids in tannase-treated whole rye flour (RFT) were higher than RF after in vitro gastrointestinal digestion, and higher amounts of transported vanillic acid through the Caco-2 monolayer were detected in RFT. However, the bioaccessibility and the transport efficiency of RF phenolic acids were higher than RFT. Underutilized crops like rye and rye-derived products may be an important source of phenolic acids. The tannase biotransformation, even influencing the total phenolics and antioxidant capacity of RF, did not increase the bioaccessibility of phenolic acids under the experimental conditions of this study.

Biotransformed citrus extract as a source of anti-inflammatory polyphenols: Effects in macrophages and adipocytes.

Chronic non-communicable diseases such as obesity are preceded by increased macrophage infiltration in adipose tissue and greater secretion of pro-inflammatory cytokines. We evaluated the anti-inflammatory potential of Biotransformed extract, and two control extracts: In Natura and Autoclaved. The assays were performed using a cellular model with RAW264.7, 3T3-L1 cells, and RAW264.7 and 3T3-L1 co-culture. The innovation of the study was the use of Biotransformed extract, a unique phenolic extract of a bioprocessed citrus residue. LPS stimulated RAW264.7 cells treated with the Biotransformed extract exhibited lower secretion of TNF-α and NO and lower protein expression of NFκB. In RAW264.7 and 3T3-L1 co-culture, treatment with 1.0mg/mL of the Biotransformed extract reduced secretion of TNF-α (30.7%) and IL-6 (43.4%). Still, the Biotransformed extract caused higher increase in adiponectin in relation to control extracts. When the co-culture received a LPS stimulus, the Autoclaved extract at 1.0mg/mL reduced IL-6 and TNF-α concentrations, and raised adiponectin. However, it was noteworthy that the Biotransformed extract was also able to significantly reduce IL-6 concentration while the Natural extract was not. The Biotransformed citrus extract evaluated in this study showed anti-inflammatory activity in macrophages and in co-culture, indicating that bioprocess of citrus residue can contribute to new product development with anti-inflammatory potential.

Immobilized tannase treatment alters polyphenolic composition in teas and their potential anti-obesity and hypoglycemic activities in vitro.

The aim of this work was to assess the effect of immobilized-tannase treatment on black, green, white and mate tea components and on their bioactivities relevant to obesity. Tannase treatment caused predictable changes in polyphenol composition with substantial reduction in galloylated catechins in green, white and black tea. Mate tea, which is rich in chlorogenic acids, was much less affected by tannase treatment although some degradation of caffeoyl quinic acid derivatives was noted. The original tea samples were effective in inhibiting digestive enzymes in vitro. They inhibited amylase activity, some with IC50 values ∼70 µg mL(-1), but were much less effective against α-glucosidase. They also inhibited lipase activity in vitro and caused dose-dependent reductions in lipid accumulation in cultured adipocytes. The bio-transformed tea samples generally matched the effectiveness of the original samples but in some cases they were markedly improved. In particular, tannase treatment reduced the IC50 value for amylase inhibition for green tea and white tea by 15- and 6-fold respectively. In addition, the bio-transformed samples were more effective than the original samples in preventing lipid accumulation in adipocytes. These in vitro studies indicate that bio-transformed tea polyphenols could assist in the management of obesity through improvement in energy uptake and lipid metabolism and also indicate that biotechnological modification of natural food molecules can improve the benefits of a common beverage such as tea.

Lipase catalyzed interesterification of Amazonian patauá oil and palm stearin for preparation of specific-structured oils.

This study showed that enzymatic interesterification of Amazonian oils could be an important tool in order to produce new oils with physicochemical properties that improve the applications of these raw materials. Structured oils of Amazonian patauá oil and palm stearin using two lipases were produced in three different enzymatic systems: first, a crude lipase from the fungus Rhizopus sp (a microorganism isolated in our laboratory); second, a commercial lipase; and third, to check any synergistic effect, a mixture of both lipases (Rhizopus sp and commercial). The lipase from Rhizopus sp was specific in the incorporation of oleic acid at the sn-1,3 positions of the triacylglycerol, resulting in an oil richer in saturated fatty acid in the sn-2 position. This enzyme, produced by solid-state fermentation, even though crude, was fatty acid and positional specific and able to operate at low concentration (2.5 %, w/w). In the second enzyme system, the commercial lipase from Thermomyces lanuginosus was not specific in the tested conditions; there was no change in the distribution of saturated and unsaturated fatty acids in the three positions of the triacylglycerol profile, there was only a replacement by the type of fatty acid at the same position. In the third enzyme system, the mixture of both lipases shows no synergic effect. The structured oils retained the concentration of bioactive α- and γ- tocopherol in the three enzyme systems. Triacylglycerol classes and Thermal behavior tests indicated the formation of more homogeneous triacylglycerols, especially the mono and di-unsaturated.

Simultaneous extraction and biotransformation process to obtain high bioactivity phenolic compounds from Brazilian citrus residues.

Recent studies have pointed to a reduction in the incidence of some cancers, diabetes, and neuro-degenerative diseases as a result of human health benefits from flavanones. Currently, flavanones are obtained by chemical synthesis or extraction from plants, and these processes are only produced in the glycosylated form. An interesting environmentally friendly alternative that deserves attention regarding phenolic compound production is the simultaneous extraction and biotransformation of these molecules. Orange juice consumption has become a worldwide dietary habit and Brazil is the largest producer of orange juice in the world. Approximately half of the citrus fruit is discarded after the juice is processed, thus generating large amounts of residues (peel and pectinolytic material). Hence, finding an environmentally clean technique to extract natural products and bioactive compounds from different plant materials has presented a challenging task over the last decades. The aim of this study was to obtain phenolics from Brazilian citrus residues with high bioactivity, using simultaneous extraction (cellulase and pectinase) and biotransformation (tannase) by enzymatic process. The highest hesperetin, naringenin and ellagic acid production in the experiment were 120, 80, and 11,250 µg g(-1), respectively, at 5.0 U mL(-1) of cellulase and 7.0 U mL(-1) of tannase at 40°C and 200 rpm. Also, the development of this process generated an increase of 77% in the total antioxidant capacity. These results suggest that the bioprocess obtained innovative results where the simultaneous enzymatic and biotransformatic extracted flavanones from agro-industrial residues was achieved without the use of organic solvents. The methodology can therefore be considered a green technology.

Fermentation and enzyme treatments for sorghum

Sorghum (Sorghum bicolor Moench) is the fifth most produced cereal worldwide. However, some varieties of this cereal contain antinutritional factors, such as tannins and phytate that may form stable complexes with proteins and minerals which decreases digestibility and nutritional value. The present study sought to diminish antinutritional tannins and phytate present in sorghum grains. Three different treatments were studied for that purpose, using enzymes tannase (945 U/Kg sorghum), phytase (2640 U/Kg sorghum) and Paecilomyces variotii (1.6 X 10(7) spores/mL); A) Tannase, phytase and Paecilomyces variotii, during 5 and 10 days; B) An innovative blend made of tanase and phytase for 5 days followed by a Pv increase for 5 more days; C) a third treatment where the reversed order of B was used starting with Pv for 5 days and then the blend of tannase and phytase for 5 more days. The results have shown that on average the three treatments were able to reduce total phenols and both hydrolysable and condensed tannins by 40.6, 38.92 and 58.00 %, respectively. Phytase increased the amount of available inorganic phosphorous, on the average by 78.3 %. The most promising results concerning tannins and phytate decreases were obtained by the enzymes combination of tannase and phytase. The three treatments have shown effective on diminishing tannin and phytate contents in sorghum flour which leads us to affirm that the proposed treatments can be used to increase the nutritive value of sorghum grains destined for either animal feeds or human nutrition.

Medium composition influence on Biotin and Riboflavin production by newly isolated Candida sp

Complex B vitamins as Biotin and Riboflavin are required by living organisms, not only for growth but also for metabolite production, and the feed market classifies them as growth promoters. Since Brazil will soon be one of the world's biggest animal protein producers, feed production is a large consumer of vitamins and micronutrients. The industry requires 10 mg riboflavin/0.2 mg biotin per kilogram of feed; a ratio of 40 ~ 50:1. Although few studies have been conducted specifically on riboflavin production using factorial design and surface response method as an optimization strategy, it is a common practice in biotechnology with many research reports available. However, there are no reports on the use of statistical design for biotin production. This study set out to evaluate medium composition influence on biotin and riboflavin production using a statistical design. There are no studies relating biotin and riboflavin production by Candida sp LEB 130. In this preliminary study to improve the simultaneous production of biotin and riboflavin, the maximum riboflavin/biotin ratio of 8.3 µg/mL was achieved with medium component concentrations of: sucrose 30 g/L, KH2PO4 2 g/L, MgSO4 1 g/L and ZnSO4 0.5mL/L.

Inoculum padronization for the production of cutinase by Fusarium oxysporum

Cutinase is a versatile enzyme showing several interesting properties for application in industrial processes. The widespread use of this enzyme depends on the development of an efficient and low-cost production system. One of the most important steps in a fermentation process is the standardization of the inoculum characteristics. In this study, the production of cutinase by Fusarium oxysporum showed a statistically significant relationship with both the inoculum size and the inoculum PDA pH. The greatest activities were 19.1 U/mL at PDA pH 7.0 and 22.72 U/mL using an aliquot of 12.72 x 10(7) spores/mL. The macroscopic characteristics of the colonies of Fusarium oxysporum changed according to the variation of the medium pH, with the best results recorded in those colonies presenting a cotton white aspect.

Cutinase é uma enzima versátil, que apresenta propriedades interessantes para aplicação em processos industriais. O uso desta enzima em larga escala depende do desenvolvimento de um sistema de produção eficiente e de baixo custo. Uma das etapas mais importantes em um processo de fermentação é a padronização do inóculo. Neste estudo, houve uma associação estatisticamente significativa entre a produção de cutinase por Fusarium oxysporum e tamanho do inóculo e pH do meio PDA. As maiores atividades de cutinase foram 19,1 U/mL em PDA com pH 7,0 e 22,72 U/mL empregando um inóculo de 12,72 x 10(7) esporos/mL. As características macroscópicas das colônias de Fusarium oxysporum mostraram alterações em função do pH do meio, com as maiores atividades sendo registradas em presença de colônias brancas com aspecto cotonoso.

A rapid screening method for cutinase producing microorganisms

A cutinase pertence, como as lípases, ao grupo das esterases (EC 3.1.1.X), que são enzimas capazes de catalisar reações de hidrólise de ligações do tipo éster. A cutinase catalisa a hidrólise da cutina, um biopoliéster insolúvel que constitui o componente estrutural da cutícula das plantas. Esta enzima tem se mostrado um eficiente catalisador tanto em solução aquosa quanto em meios orgânicos, sendo potencialmente apropriada para usos em indústria de detergentes, alimentos e cosméticos. O objetivo deste trabalho foi isolar fungos de diversas fontes e realizar uma pré-seleção destes, quanto à habilidade em produzir esterase. As linhagens fúngicas pré-selecionadas como produtoras de esterase foram inoculadas em meio de cultivo contendo cutina extraída de maçã. As atividades cutinolítica e lipolítica foram medidas no sobrenadante das culturas a fim de diferenciar os microrganismos produtores de lipase dos produtores de cutinase. Uma linhagem isolada mostrou a maior atividade cutinolítica após 12 dias de fermentação em um meio contendo 1 por cento de cutina e foi identificada como sendo o Fusarium oxysporium.