Structural properties and evaluation of the antiproliferative activity of limonene-1,2-diol obtained by the fungal biotransformation of R-(+)- and S-(-)-limonene.

Limonene-1,2-diol is a limonene oxygenated metabolite that possesses eight different stereoisomers, which could result in different biological properties. Nonetheless, the relation between its spatial configuration and biological function is still little explored. The present study aimed to perform the stereoisomers identification using nuclear magnetic resonance (NMR) investigation of the limonene-1,2-diol produced via R-(+)- and S-(-)-limonene biotransformation by Colletotrichum nymphaeae and S-(-)-limonene biotransformation by Fusarium oxysporum 152B. Besides, in vitro antiproliferative activity was evaluated against human tumor and nontumor cell lines. The NMR analysis showed that R-(+)-limonene biotransformation afforded exclusively (+)-(1S,2S,4R-limonene-1,2-diol), whereas S-(-)-limonene biotransformation afforded exclusively (-)-(1R,2R,4S-limonene-1,2-diol) independent on the fungi used. Despite no significant cytostatic effects, a possible influence of stereogenic center on the antiproliferative activity of these limonene biotransformation products was evidenced. Moreover, the lack of in vitro antiproliferative effect of limonene-1,2-diol against nontumor cells suggested a safe dose range for further in vivo evaluations, including food applications.

Relation of shear stress and KLa on bikaverin production by Fusarium oxysporum CCT7620 in a bioreactor.

This study involved evaluating the effects of rotational impeller speed agitation (N) and specific air flow rate (Фair) on bikaverin production and on the growth of Fusarium oxysporum employing 11 bench-scale bioreactor assays. The results showed that the maximum bikaverin production (close to 300 mg L-1) was achieved after 48 h of fermentation in rice medium (20 g L-1 milled rice in water) at 28 °C with a volumetric oxygen transfer coefficient (KLa) and shear stress values of approximately 20 h-1 and 17 N m-2, respectively. We reached this combination of parameters using an N of 340 rpm and Фair of 0.935 vvm. These KLa and shear stress values can be used as references when upscaling this process. Thus, this study was important to demonstrate how the main parameters in bioreactors affect bikaverin production and it presented important indications for upscaling this bioprocess.

Pigments from Antarctic bacteria and their biotechnological applications.

Pigments from microorganisms have triggered great interest in the market, mostly by their "natural" appeal, their favorable production conditions, in addition to the potential new chemical structures or naturally overproducing strains. They have been used in: food, feed, dairy, textile, pharmaceutical, and cosmetic industries. The high rate of pigment production in microorganisms recovered from Antarctica in response to selective pressures such as: high UV radiation, low temperatures, and freezing and thawing cycles makes this a unique biome which means that much of its biological heritage cannot be found elsewhere on the planet. This vast arsenal of pigmented molecules has different functions in bacteria and may exhibit different biotechnological activities, such as: extracellular sunscreens, photoprotective function, antimicrobial activity, biodegradability, etc. However, many challenges for the commercial use of these compounds have yet to be overcome, such as: the low stability of natural pigments in cosmetic formulations, the change in color when subjected to pH variations, the low yield and the high costs in their production. This review surveys the different types of natural pigments found in Antarctic bacteria, classifying them according to their chemical structure. Finally, we give an overview of the main pigments that are used commercially today.

Lipase production by microorganisms isolated from the Serra de Ouro Branco State Park.

Lipases are hydrolases used in various sectors such as the food, pharmaceutical and chemical synthesis industries. In this study, epiphytic microorganisms were isolated from the Serra of Ouro Branco State Park (Minas Gerais, Brazil) and were subsequently evaluated for their ability to produce extracellular lipases. Among the 46 isolated strains, 25 presented positive results for lipase production in the agar plate screening assay. Two of these strains that expressed the highest diffusion halos, were genetically identified as Serratia marcescens and Pseudomonas fluorescens and catalogued in the Tropical Cultures Collection from the André Tosello Foundation/Brazil as CCT 7796 and CCT 7797, respectively. The fermentation growth kinetics indicated that the maximum extracellular lipase activities were achieved between 96 and 120h of cultivation. The highest lipolytic activity for both strains was observed at an optimum temperature and pH of 37°C and 7.0, respectively. At these conditions, the lipase activity detected in the crude enzymatic extract of both strains was close to 15.0 U/mL. We consider that these species are promising lipase producers for industrial applications.

Comparison of Two Methods for Counting Molds in Fermentations Using the Production of Bikaverin by Fusarium oxysporum CCT7620 as a Model.

The dry cell weight (DCW) measurement is one of the preferred methods to determine the growth of filamentous fungi. However, this technique is not applicable to insoluble culture media, besides being possibly influenced by the presence of extracellular biomass. The standard plate counting (SPC) is a reference method for detecting viable cells; however, it is referred as imprecise. In this study, we did a comprehensive analysis of the errors associated to each procedure and also determined the growth kinetics of Fusarium oxysporum in soluble (DCW and SPC) and insoluble (SPC) culture media. Finally, we used the production of bikaverin in airlift bioreactor containing insoluble medium as a case study to estimate red pigment production and to monitor biomass growth via SPC. We concluded that SPC can be used to give reliable fungal growth kinetics in media with insoluble matter, yielding errors equivalent to DCW depending on the number of replicates done for serial dilutions and plate counting.

Monoterpene biotransformation by Colletotrichum species.

OBJECTIVE: To investigate the biocatalytic potential of Colletotrichum acutatum and Colletotrichum nymphaeae for monoterpene biotransformation. RESULTS: C. acutatum and C. nymphaeae used limonene, α-pinene, ß-pinene, farnesene, citronellol, linalool, geraniol, perillyl alcohol, and carveol as sole carbon and energy sources. Both species biotransformed limonene and linalool, accumulating limonene-1,2-diol and linalool oxides, respectively. α-Pinene was only biotransformed by C. nymphaeae producing campholenic aldehyde, pinanone and verbenone. The biotransformation of limonene by C. nymphaeae yielded 3.34-4.01 g limonene-1,2-diol l-1, depending on the substrate (R-(+)-limonene, S-(-)-limonene or citrus terpene (an agro-industrial by-product). This is among the highest concentrations already reported for this product. CONCLUSIONS: This is the first report on the biotransformation of these terpenes by Colletotrichum spp. and the biotransformation of limonene to limonene-1,2-diol possibly involves enzymes similar to those found in Grosmannia clavigera.

Lipase-mediated alcoholysis for in situ production of ester bioaromas in licuri oil for cosmetic applications.

Bioaromas can be produced by lipases either through their hydrolytic or (trans)esterifying activities. Therefore, this work reports the development of a lipase-catalyzed biotransformed licuri oil, forming volatile ethyl esters with odor notes resembling tropical fruits. Ethyl octanoate formation was promoted when 7.0 % (m/v) Lipozyme 435® was used to convert a grain alcohol:licuri oil mixture (51:49, v/v) at 58ºC and 70 rpm for 6 hours. The biotransformed oil has shown antimicrobial activity against Staphylococcus hominis, S. epidermidis, and Corynebacterium xerosis, bacteria associated with bad skin odor. Finally, this biotransformed oil was used without further treatments (e.g., recovery or purification procedures) to prepare two cosmetic formulations (in a dosage of 1.5 %), aiming for both fragrant and deodorant activity.

Antifungal activity of cercosporamide produced by Phaeosphaeriaceae GV-1 against pathogenic fungi.

Fungal infections affect millions of people worldwide, and the several cases are related to invasive infections, which is a problem mainly for immunocompromised people, such as transplant and cancer patients with high mortality and morbidity rates. In addition, the number of emerging and multidrug-resistant fungal species has increased in the last decade. The search for new antifungal compounds is necessary, due to the increase in cases of resistance and the toxicity of drugs used in fungal infection treatment. This work aimed to study the antifungal activity of cercosporamide produced by Phaeosphaeriaceae GV-1. Cercosporamide was tested against pathogenic fungi by determining the minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations, using the broth microdilution method. Cercosporamide showed antifungal activity in vitro against 13 of 16 strains of medical importance tested, with the most susceptible species being Candida tropicalis, with MIC and MFC of 15.6 µg/mL. Thus, cercosporamide might be considered a promising therapeutic antifungal agent.

Supercritical CO2 as a Valuable Tool for Aroma Technology.

This review addresses the possibilities of using supercritical carbon dioxide (SC-CO2) in the flavor industry in extraction and fractionation processes and its use as a reaction medium to generate aroma esters. The advantages and disadvantages are presented, comparing SC-CO2 processing with traditional methods. The most distinguishable features of SC-CO2 include mild reaction conditions, time savings, fewer toxicity concerns, higher sustainability, and the possibility of modulating solvent selectivity according to the process conditions (such as pressure and temperature). Thus, this review indicates the potential of using SC-CO2 to obtain a high selectivity of compounds that can be applied in aroma technology and related fields.

Lignocellulosic substrates as starting materials for the production of bioactive biopigments.

The search for sustainable processes is constantly increasing in the last years, so reusing, recycling and adding value to residues and by-products from agroindustry is a consolidated area of research. Particularly in the field of fermentation technology, the lignocellulosic substrates have been used to produce a diversity of chemicals, fuels and food additives. These residues or by-products are rich sources of carbon, which may be used to yield fermentescible sugars upon hydrolysis, but are usually inaccessible to enzyme and microbial attack. Therefore, pre-treatments (e.g. hydrolysis, steam explosion, biological pretreatment or others) are required prior to microbial action. Biopigments are added-value compounds that can be produced biotechnologically, including fermentation processes employing lignocellulosic substrates. These molecules are important not only for their coloring properties, but also for their biological activities. Therefore, this paper discusses the most recent and relevant processes for biopigment production using lignocellulosic substrates (solid-state fermentation) or their hydrolysates.

Formulation and physicochemical stability of oil-in-water nanoemulsion loaded with α-terpineol as flavor oil using Quillaja saponins as natural emulsifier.

Alpha-terpineol (α-TOH) is a promising monoterpenoid detaining several biological activities. However, as a volatile molecule, the incorporation of α-TOH within formulated products poses several challenges related to its stability. In this sense, nanoencapsulation works as a key technology to protect the bioactivity of low molecular weight oils, like α-TOH, against environmental stresses (heat, light, and moisture), mitigating their susceptibility to degradation (oxidation and volatilization). Physical properties of encapsulated flavor/essential oil have been extensively reported, whereas there is a lack in the literature regarding their chemical stability, which is usually the main purpose of encapsulation. Thus, in this study, the physicochemical stability of the formulated oil-in-water nanoemulsion loaded with α-TOH stabilized with Quillaja saponins (QSs) as a natural emulsifier (α-TOH-QSs-NE) were tracked in a long-term (up to 280th day). Along with time, mean droplet diameter (MDD) and turbidity were used as a reference for physical parameters; while the chemical stability was monitored using gas chromatography analysis to quantify the mark content of α-TOH into the NE. Results indicated that α-TOH-QSs-NE was successfully formulated with a high-load amount of α-TOH (90 mg mL-1). α-TOH-QSs-NE showed great physicochemical stability regardless the storage-temperature (5 °C or 25 °C) up to 280th day, with no significant alterations in the MDD or turbidity, where c.a. 79% of the initial amount of the nanoemulsified α-TOH remained detectable in α-TOH-QSs-NEs, with no finding of degradation products. Thus, the data here disclosure may be useful for innovative application of α-TOH in foodstuffs.

Effect of supercritical CO2 fractionation of Tahiti lemon (Citrus latifolia Tanaka) essential oil on its antifungal activity against predominant molds from pan bread.

This study aimed to verify the in vitro antifungal activity of Tahiti lemon essential oil (LEO) and its fractions, obtained by supercritical CO2 fractionation, against Penicillium sumatrense and Aspergillus niger isolated from pan bread. For this, LEO was solubilized (20 MPa and 40 °C) and fractionated (10 MPa and 40 °C) in supercritical CO2, resulting in soluble (SF) and precipitated (PF) fractions. LEO, SF and PF volatile compounds were identified by gas chromatography coupled to mass spectrometry (GC-MS) and semiquantified by gas chromatography with a flame ionization detector (GC-FID). To evaluate the in vitro antifungal activity of the essential oils (LEO, SF and PF), the minimal inhibitory and fungicidal concentrations (MIC and MFC, respectively) were determined using the 96-well plate methodology. For this, pan breads ware prepared with no preservatives and stored for seven days at 25 °C, and their pH, water activity and moisture contents were determined. Then, two predominant species (Penicillium sumatrense and Aspergillus niger) were isolated from pan breads, characterized according to their morphological and molecular characteristics, and were used in the antifungal activity studies. LEO and its fractions presented monoterpenes, sesquiterpenes and their oxygenated derivatives in their composition. Specifically, limonene was the major component identified in the essential oils. SF showed greater antifungal potential than PF and LEO, showing that supercritical CO2 fractionation could improve the antifungal efficiency of LEO. The results suggest that LEO and its fractions may contribute to the inhibition of Aspergillus niger and Penicillium sumatrense growth in pan breads.

Natural blue pigments and bikaverin.

In last years, the main studied microbial sources of natural blue pigments have been the eukaryotic algae, Rhodophytes and Cryptophytes, and the cyanobacterium Arthrospira (Spirulina) platensis, responsible for the production of phycocyanin, one of the most important blue compounds approved for food and cosmetic use. Recent research also includes the indigoidine pigment from the bacteria Erwinia, Streptomyces and Photorhabdus. Despite these advances, there are still few options of microbial blue pigments reported so far, but the interest in these products is high due to the lack of stable natural blue pigments in nature. Filamentous fungi are particularly attractive for their ability to produce pigments with a wide range of colors. Bikaverin is a red metabolite present mainly in species of the genus Fusarium. Although originally red, the biomass containing bikaverin changes its color to blue after heat treatment, through a mechanism still unknown. In addition to the special behavior of color change by thermal treatment, bikaverin has beneficial biological properties, such as antimicrobial and antiproliferative activities, which can expand its use for the pharmaceutical and medical sectors. The present review addresses the production natural blue pigments and focuses on the properties of bikaverin, which can be an important source of blue pigment with potential applications in the food industry and in other industrial sectors.

Delaying crystallization in single fractionated palm olein with limonene addition.

Single fractionated palm olein (OL) becomes cloudy when submitted to low temperatures. To overcome this technological issue, the use of appropriate additives delays or prevents its clouding. Limonene is considered a green additive, and studies revealed that it modulates fat crystallization. This study evaluated the influence of adding R-(+)-limonene, in different concentrations (1-10%), into OL, regarding its crystallization behavior. The findings show that addition of limonene reduced solid fat content (SFC), crystallization temperature by differential scanning calorimetry (DSC), and cloud point of OL, and the results were more pronounced at higher concentrations of limonene. The blend OL + 10% limonene presented the best resistance in cold stability. From the obtained results, the blends fitted as intermediate products between an OL and a super palm olein (SOL), with substantial improvement in reducing crystals' formation in OL. Limonene can be considered a green anti-crystallizer with potential application in different areas, such as cosmetics and biodiesel.

Comprehensive study of α-terpineol-loaded oil-in-water (O/W) nanoemulsion: interfacial property, formulation, physical and chemical stability.

In this study, the interfacial ability of α-terpineol (α-TOH) was reported, followed by its trapping into oil-in-water (O/W) nanoemulsion as active-ingredient and the long-term observation of this nanosystem influenced by the storage-time (410-days) and temperature (5, 25, 50 °C). The results indicated that the α-TOH can reduce the interfacial tension on the liquid-liquid interface (ΔG°m = -1.81 KJ mol-1; surface density = 8.19 × 10-6 mol m-2; polar head group area = 20.29 Å2), in the absence or presence of surfactant. The O/W nanoemulsion loaded with a high amount of α-TOH (90 mg mL-1; 9α-TOH-NE) into the oil phase was successfully formulated. Among the physical parameters, the mean droplet diameter (MDD) showed a great thermal dependence influenced by the storage-temperature, where the Ostwald ripening (OR) was identified as the main destabilizing phenomena that was taking place on 9α-TOH-NE at 5 and 25 °C along with time. Despite of the physical instability, the integrity of both nanoemulsion at 5 °C and 25 °C was fully preserved up to 410th day, displaying a homogeneous and comparable appearance by visual observation. On contrary, a non-thermal dependence was found for chemical stability, where over 88% of the initial amount of the α-TOH nanoemulsified remained in both 9α-TOH-NE at 5 and 25 °C, up to 410th day. Beyond the key data reported for α-TOH, the importance of this research relies on the long-term tracking of a nanostructured system which can be useful for scientific community as a model for a robust evaluation of nanoemulsion loaded with flavor oils.

Recovery and purification of bikaverin produced by Fusarium oxysporum CCT7620.

Microbial pigments have a distinguished potential for applications in food and pharmaceutical industries, stimulating the research in this field. The present study evaluated the ideal conditions for extracting bikaverin (red pigment) from the biomass of Fusarium oxysporum CCT7620. Among the solvents tested, ethyl acetate extraction resulted in the highest bikaverin concentration and the kinetic study revealed a saturation in bikaverin concentration from 256 min on. Based on a preliminary economic study, three sequential extractions with ethyl acetate was considered the ideal protocol to recover bikaverin. After extraction, chromatographic methods were tested to purify bikaverin. The use of silica gel or Sephadex (open column) could not successfully purify bikaverin, but the semi-preparative HPLC resulted in a bikaverin-enriched fraction with a purity degree equivalent to the commercial analytical standard. This work provides relevant information regarding the extraction and purification of bikaverin, which may be useful for other downstraming processes.

Non-nutrients and nutrients from Latin American fruits for the prevention of cardiovascular diseases.

Non-communicable diseases (NCDs) have been rapidly increasing; among them, cardiovascular diseases (CVDs) are responsible for around 1/3 of deaths in the world. Environmental factors play a central role in their development. Diet is a very important factor in this scenario, and the intake of fruits and vegetables has been considered as one of the critical strategies for reducing the risk of CVDs. Fruits are a source of micronutrients and bioactive compounds that could have cardioprotective effects through several distinct mechanisms, such as antioxidant, antithrombotic and antiplatelet activities, vasodilatation, improvement of plasma lipid profiles, and modulation of inflammatory signaling. Brazil has a very rich and unexplored biodiversity in its different biomes, with several types of fruit, which are a source of bioactive compounds and micronutrients with therapeutic properties. In this sense, this review shows the current knowledge regarding the cardioprotective properties of selected Latin American and Brazilian fruits, including their effects on the activation of platelets and on the inflammation processes involved in atherosclerosis and cardiovascular diseases.

Skin microbiota as a therapeutic target for psoriasis treatment: Trends and perspectives.

Psoriasis is a chronic, immune-mediated disease that has a major negative impact on a patient's quality of life. Although several literature reviews indicate that skin microbiota may play an important role in the development and regulation of the immune and inflammatory response of psoriasis, few clinical studies are demonstrating the benefits of using pre-, pro- and synbiotics as a therapeutic alternative at the management of the disease. In this review, we showed the use of probiotic microorganisms that may contribute to skin homeostasis and compiled the clinical trials that demonstrate the effect of therapy with probiotics on patients with psoriasis, an important area for scientific exploration in dermatology and being the first review article to compile this information.

The effect of α-terpineol enantiomers on biomarkers of rats fed a high-fat diet.

Alpha-terpineol is a monoterpenoid found in many essential oils, being widely used in food and household products. In vitro antioxidant and anti-inflammatory activities have already been associated with this alcohol; therefore, this study aimed to check if these properties were also present in vivo, counteracting the oxidant and inflammatory effects of a high-fat diet, as well as if there were differences in the biological activities among the two α-terpineol enantiomers. Thus, this work evaluated the effect of supplementation of α-terpineol enantiomers (at 25, 50 and 100 mg/kg of diet) on biological parameters of diet-induced obese Sprague-Dawley rats. In general, α-terpineol improved the nutritional parameters of rats fed a high-fat diet. The intake of α-terpineol at concentrations ≥50 mg/kg was able to reestablish the insulin sensibility and reduced (p < 0.05) serum levels of pro-inflammatory cytokines TNF-α and IL-1ß, when compared with the control group. The intake of R-(+)- and (-)-α-terpineol decreased the TNF-α level by approximately 1.5 and 3.4 times, respectively, when compared with the high-fat group, regardless of the concentration. Moreover, both enantiomers at 50 mg/kg decreased the levels of serum thiobarbituric acid reactive substances (TBARS) by 2.6-4.2 times, while hepatic TBARS were reduced in approximately 1.6 times, regardless of the compound and concentration tested. Further experiments are suggested to confirm the mechanisms and the security of α-terpineol in different experimental models and more extended exposure experiments.

Optimization of limonene biotransformation for the production of bulk amounts of α-terpineol.

The biotransformation of R-(+)-limonene into high concentrations of R-(+)-α-terpineol by Sphingobium sp. was investigated in order to optimize the main process variables (pH, biocatalyst concentration, substrate concentration, temperature and agitation). This strategy comprised the screening of variables by a Plackett-Burman design followed by a Central Composite Design. The statistical analysis showed that the optimal α-terpineol production were at 28 °C and pH 7.0, with a limonene concentration of 350 g/L of organic phase agitation of 200 rpm and a biocatalyst concentration of 2.8 g/L of aqueous phase (OD600 = 8). Further trials showed that the R-(+)-α-terpineol concentration was higher (240 g/L after 96 h) when using a ratio of 1:3 (v.v-1) of organic:aqueous phases. However, the total production and yield (in terms of biomass) of α-terpineol would be maximized for an aqueous:organic ratio of 1:1. The experimental design optimization adopted herein was an effective tool for this type of study.