Researching New Drug Combinations with Senolytic Activity Using Senescent Human Lung Fibroblasts MRC-5 Cell Line.

Therapeutically targeting senescent cells seems to be an interesting perspective in treating chronic lung diseases, which are often associated with human aging. The combination of the drug dasatinib and the polyphenol quercetin is used in clinical trials as a senolytic, and the first results point to the relief of physical dysfunction in patients with idiopathic pulmonary fibrosis. In this work, we tested new combinations of drugs and polyphenols, looking for senolytic activity using human lung fibroblasts (MRC-5 cell line) with induced senescence. We researched drugs, such as azithromycin, rapamycin, metformin, FK-506, aspirin, and dasatinib combined with nine natural polyphenols, namely caffeic acid, chlorogenic acid, ellagic acid, ferulic acid, gallic acid, epicatechin, hesperidin, quercetin, and resveratrol. We found new effective senolytic combinations with dasatinib and ellagic acid and dasatinib and resveratrol. Both drug combinations increased apoptosis, reduced BCL-2 expression, and increased caspase activity in senescent MRC-5 cells. Ellagic acid senolytic activity was more potent than quercetin, and resveratrol counteracted inflammatory cytokine release during senolysis in vitro. In conclusion, dasatinib and ellagic acid and dasatinib and resveratrol present in vitro senolytic potential like that observed for the combination in clinical trials of dasatinib and quercetin, and maybe they could be future alternatives in the senotherapeutic field.

Effect of Enzymatic Biotransformation on the Hypotensive Potential of Red Grape Pomace Extract.

Hypertension is a widespread health risk, affecting over a billion people and causing 9 million deaths per year. The Renin-Angiotensin-Aldosterone System (RAAS) is a primary target for hypertension treatment, and it is primarily treated through drugs that inhibit the Angiotensin I-Converting Enzyme (ACE). In addition to pharmacological treatment, various plants are recommended in traditional medicine for blood pressure regulation. This study aimed to produce high-phenolic-content extracts with and without enzymatic assistance from red grape pomace and evaluate their antioxidant capacity and ACE inhibitory potential. The total phenolic content (TPC) was measured, and phenolic identification was performed using HPLC analysis. In addition, the antioxidant capacity and anti-hypertensive potential were determined via in vitro assays. There was no statistical difference in the TPC antioxidant capacity between the extraction methods. Otherwise, when considering the extraction yield, the enzymatic process recovered around 70% more phenolic compounds from the pomace, and the phenolic profile was changed. Enzymatic assistance also significantly increased the ACE inhibitory potential in the grape pomace extract. This study demonstrates the viability of upcycling grape pomace to obtain bioactive compounds and to reduce their environmental impact, and highlights the influence of the enzymatic extraction on the hypotensive potential of the extract.

Production and characterization of nanoemulsion with low-calorie structured lipids and its potential to modulate biomarkers associated with obesity and comorbidities.

Structured lipids (SL) containing behenic acid have been produced in order to obtain low-calorie lipids for foods; however, the development of a high nutritional value and a stable nanoemulsion carrier system for these SL is an interesting breakthrough for this field of research, improving technologic and biological potential for food application. In this sense, the aim of this study was to evaluate the stability of a nanoemulsion containing SL NeSL (produced with olive oil, soybean oil and fully hydrogenated crambe oil), the behavior during in vitro digestion and the effects on biomarkers involved in the obesity in cell models. The samples showed good stability throughout storage (30 days) under refrigeration and room temperature and after the gastric digestion phase compared to the controls (nanoemulsion of olive and soybean oil). After the intestinal phase, there was an increase in oil droplet size and zeta potential, a characteristic of coalescence. In the lipid accumulation model in adipocytes, the highest concentration (50 µL/mL) of NeSL resulted in 42% less lipid accumulation, compared to the control. Furthermore, the sample was able to reduce inflammatory cytokines produced by macrophages provoked by LPS (lipopolysaccharide). The combination of the oils in NeSL resulted in a fatty acid profile with beneficial health properties, which may have contributed to less lipid accumulation and improved inflammatory parameters. This SL in the form of a nanoemulsion, may be used as a partial fat substitute in low-calorie food products.

Effects of enzyme-assisted extraction on the profile and bioaccessibility of isoflavones from soybean flour.

The effects of enzymatic extraction strategies on extractability, bioconversion, and bioaccessibility of biologically active isoflavone aglycones, total phenolic content, and antioxidant activity of aqueous extracts from full-fat soy flour were evaluated. Protease, tannase, and cellulase enzymes were used individually or in combination. Except for the protease treatment, all enzymatic treatments increased the extraction of biologically active isoflavones (daidzein and genistein) compared with the control. The use of a mixture of protease, tannase, and cellulase resulted in increased extractability and/or bioconversion of aglycones from soy flour, indicating a synergistic effect amongst the enzymes. Daidzein and genistein concentrations increased from 29.0 to 158.2 µg/g and from 27.0 to 156.5 µg/g (compared to the control), respectively. Furthermore, enzymatic extraction followed by in vitro gastrointestinal digestion significantly increased the bioaccessibility of isoflavone aglycones, total phenolic content (by 22-45%), and antioxidant activity (by 15-22%) of the extracts. These results demonstrate that enzyme selection is an efficient strategy to maximize the extraction, bioconversion, and bioaccessibility of bioactive isoflavones from soy flour, which could contribute to health benefits associated with the consumption of soy-rich products.

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.

Biotransformation processes in soymilk isoflavones to enhance anti-inflammatory potential in intestinal cellular model.

The present study investigated, in in vitro cellular model, the modulation of intestinal inflammation by biotransformed soymilk with tannase and probiotic strains. The ability to reduce the generation of intracellular reactive oxygen species (ROS) and the antioxidant power of soy extracts were also evaluated. The results showed changes in isoflavones profile after biotransformation processes, with a significative enhancement in aglycones content. Reduction in intracellular ROS production and improvement in antioxidant capacity were observed. Anti-inflammatory responses in Caco-2 cells were also expressive. A significative decrease in interleukin 8 (IL-8) level was detected for all biotransformed samples, especially for extracts with tannase. The biotransformed soy extracts by tannase have a great potential to improve health conditions, defending the intestinal cells of oxidative damage, and acting as a possible adjuvant in inflammatory process. PRACTICAL APPLICATIONS: Soy isoflavones have been explored owing to health benefits. Only glycosylated forms are found in high concentrations in soybeans. So, microbial and enzymatic biotransformation processes aiming to increase aglycones and metabolites appear as an attractive option to enlarge the bioactivity of soy products. The present study showed a positive impact of biotransformed soymilk on antioxidant defenses systems and modulation of intestinal inflammation and could act as a nutraceutical agent.

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.

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.

A new biotechnological process to enhance the soymilk bioactivity.

Equol, a daidzein metabolite produced exclusively by intestinal bacteria in some, but not all, humans, exhibits a wide range of beneficial health effects owing to its superior nutraceutical effect compared with isoflavones of soy. The aim of this work was to develop bioprocesses capable of increasing the bioactive properties of soymilk and, most importantly, increase the equol content by a biotechnological process in vitro. Biotransformation processes based on soymilk fermentation by probiotic lactic bacteria and application of the enzyme tannase caused an increase in the bioactive isoflavones and antioxidant capacity of soymilk. Furthermore, these processes approximately resulted in a 10-fold increase in the equol content of the soymilk. This is the first study to produce a significant equol concentration in soymilk using enzymatic processing only. The results suggest a new and effective biotechnological process, with major commercial potential, capable of producing a bioactive soy extract that intends to be "functional for everyone."

Biotransformation effects on anti lipogenic activity of citrus extracts.

Citrus peel is a good source of flavonoids, with higher content in relation to pulp. This study proposed to investigate the anti-lipogenic potential of a newly developed citrus flavonoids extract, obtained from citrus industrial residue, bioprocessed in order to generate a commercial source of some flavonoids naturally found in low quantity. The results showed that the citrus peel extract obtained after biotransformation was a good source of hesperitin and naringenin, flavonoids that has no source for production on a large scale, as in supplements or medicines. Still, the results showed that all extracts could be used in obesity treatment. The original extract, "In Natura", would be useful to reduce new adipocytes synthesis and lipid accumulation, and the extract bioprocessed, "Biotransformed" extract could be used to induce lipolysis on fat tissue.

Seletividade de inseticidas a alguns dos inimigos naturais na cultura do algodão / Selectivity of insecticides to some natural enemies in the culture of cotton

O objetivo foi estudar a seletividade de inseticidas aos inimigos naturais na cultura do algodão (Gossypium hirsutum L.r. latifolium Hurtch), cultivar DeltaOpal, no município de Malhada (BA) e conhecer a fauna benéfica associada. O trabalho foi conduzido na safra 2010/2011, com delineamento experimental em blocos ao acaso, com seis tratamentos e quatro repetições, sendo os tratamentos: (1) Fipronil 200 SC (0,38 L.ha-1); (2) Alfacipermetrina 100 SC (0,30 L.ha-1); (3) Lufenuron 50 CE (0,30 L.ha-1); (4) Imidacloprid 200 SC (0,30 L.ha-1); (5) Paration Metil 600 CE (1,00 L.ha-1); e (6) Testemunha (água). A aplicação dos produtos foi feita 80 dias após a emergência, com avaliações um dia antes da aplicação e 1, 7 e 14 dias após a aplicação (DAA) por meio do pano de batida e armadilha Moericke. Em laboratório, os insetos foram triados, contados e identificados em famílias. A toxicidade dos produtos varia em função do grupo do inimigo natural. A Imidacloprida é seletiva às aranhas, sendo os demais inseticidas medianamente tóxicos (Alfacipermetrina e Paration Metil: 1º e 14º DAA; Lufenuron: 14º DAA) ou tóxicos (Fipronil e Alfacipermetrina: 7º DAA). Fipronil (1º DAA), Alfacipermetrina (7º DAA) e Paration Metil (14º DAA) são medianamente tóxicos aos adultos de joaninhas. Os inseticidas estudados são tóxicos às larvas de joaninhas, com mais impacto até os sete dias após a aplicação, com exceção do Paration Metil, classificado como inócuo até esse período. Registra-se a ocorrência de 13 famílias de aranhas e 18 famílias de hymenópteros parasitoides em agroecossistema de algodão na região de Malhada (BA).(AU)

The aim of this work was study the selectivity of insecticides in favor of natural enemies in cotton (Gossypium hirsutum latifolium Hurtch Lr), DeltaOpal cultivar, in the city of Malhada (BA), and to know the associated beneficial fauna. The study was conducted at the agricultural year of 2010/2011. The design was conducted in randomized blocks with six treatments and four replications. The treatments were: (1) Fipronil 200 SC (0.38 L.ha-1); (2) Alphacypermethrin 100 SC (0.30 L.ha-1), (3) Lufenuron 50 EC (0.30 L.ha-1), (4) Imidacloprid 200 SC (0.30 L.ha-1), (5) Methyl parathion 600 EC (1.00 L.ha-1), and (6) control (water). The product was applied 80 days after emergence, and the evaluations were performed one day before application and 1, 7 and 14 days after application (DAA). The samples were taken using the sampling method beating cloth and Moericke traps. Natural enemies were brought to the laboratory for sorting, counting and identification by family. The toxicity of the products ranged according to the group of natural enemies. Imidacloprid is selective to the spiders and insecticides are moderately toxic (Methyl Parathion and Alphacypermethrin: 1 and 14th DAA; Lufenuron: 14th DAA) or toxic (Fipronil and Alphacypermethrin: 7thDAA). Fipronil (1 DAA), Alphacypermethrin (7th DAA) and Methyl Parathion (14th DAA) are moderately toxic to adult ladybirds. The analyzed insecticides are toxic to the larvae of ladybirds, with more impact until seven days after the application, with the exception of Methyl Parathion classified, as innocuous until this period. The occurrence of 13 families of spiders and 18 families of parasitic Hymenoptera is registered in cotton agroecosystems in the region of Malhada, in the state of Bahia.(AU)

A new process for simultaneous production of tannase and phytase by Paecilomyces variotii in solid-state fermentation of orange pomace.

The production of enzymes such as tannases and phytases by solid-state fermentation and their use in animal feed have become a subject of great interest. In the present work, Paecilomyces variotii was used to produce tannase and phytase simultaneously. Solid-state fermentation, a process initially designed for tannase production, was implemented here using orange pomace as substrate. Orange pomace is the waste product of the large orange juice industry in Brazil, and it has also been used as an ingredient in animal feed. In addition to enzymatic production, biotransformation of the phenolic content and antioxidant capacity of the orange pomace were analyzed after fermentation. Fermentation conditions, namely moisture level and tannic acid concentration rate, were studied using CCD methodology. The response surface obtained indicated that the highest tannase activity was 5,000 U/gds after 96 h at 59% (v/w) and 3% (w/w) and that of phytase was 350 U/gds after 72 h at 66% (v/w) and 5.8% (w/w) of moisture level and tannic acid concentration, respectively. The amount of tannase production was similar to the levels achieved in previous studies, but this was accomplished with a 7% (w/w) reduction in the amount of supplemental tannic acid required. These results are the first to show that P. variotii is capable of producing phytase at significant levels. Moreover, the antioxidant capacity of orange pomace when tested against the free radical ABTS was increased by approximately tenfold as a result of the fermentation process.

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.

Detoxification of castor bean residues and the simultaneous production of tannase and phytase by solid-state fermentation using Paecilomyces variotii.

In this work, we introduce a biological detoxification method that converts toxic waste from castor beans into animal feed material. This method simultaneously induces the production of tannase and phytase by Paecilomyces variotii; both enzymes have high levels of activity and have the potential to be used in feedstuffs because they decrease overall anti-nutritional factors. The maximum tannase and phytase activities obtained were 2600 and 260 U/g after 48 and 72 h, respectively. SDS-PAGE electrophoresis of the fermented castor cake extracts revealed a reduction in ricin bands during fermentation, and the bands were no longer visible after 48 h. The cytotoxicity of the extracts was evaluated by MTT testing on RAW cells, and a progressive increase in cellular viability was obtained, reaching almost 100% after 72 h of fermentation.

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.

Optimization of medium composition for transglutaminase production by a Brazilian soil Streptomyces sp

Finding a new microbial source of transglutaminase (MTGase) and the study of the medium composition for MTGase production were the goals of this work. A total of 200 actinomycete-like strains were isolated from Brazilian soil samples and two of them named T10b and P20 were selected based on their ability to produce 0.15 U.mL-1 and 0.25 U.mL-1 of MTGase, respectively. Strain P20 was chosen to continue the study and was identified as Streptomyces sp. In order to optimize the MTGase activity, modifications of the usual media composition described for enzyme production were tested. The strategy adopted was: (1) screening experiment for the best carbon and nitrogen sources; (2) fractional factorial design (FFD) to elucidate the key ingredients in the media (the results indicated that the soybean flour, peptone, KH2PO4 and MgSO4.7H2O had a significant effect on MTGase) production and (3) central composite design (CCD) to optimize the concentration of the key components. The experimental results were fitted to a second-order polynomial model at the 95 percent level of significance (P < 0.05). Under the proposed optimized conditions, the model predicted a MTGase activity of 1.37 U.mL-1, very closely matching the experimental activity of 1.4 U.mL-1.