SELEÇÃO DE FUNGOS DO SEMI-ÁRIDO BAIANO SECRETORES DE HIDROLASES DE INTERESSE EM ALIMENTOS

Enzimas são o principal alvo da pesquisa biotecnológicadevido ao seu papel nos mecanismos celulares e a sua aplicação emprocessos industriais. A indústria de alimentos está entre as principaisconsumidoras de enzimas, pois essas são indispensáveis a processos tradicionais(cerveja) e inovadores (prebióticos). A microbiota do semiáridobaiano apresenta potencial para produção de enzimas de aplicaçãoindustrial. Este trabalho teve por objetivo selecionar fungos filamentosos,isolados do semi-árido baiano, secretores de hidrolases de interesse emalimentos, pela metodologia “cup plate”. Dos 20 fungos estudados, trêsapresentaram atividade sobre a celulose, dois apresentaram atividadesobre a pectina e 12 apresentaram atividade sobre proteínas

Analiza teobromina i srodnih spojeva pomoću visokodjelotvorne tekućinske kromatografije obrnutih faza s UV-detekcijom (Pregled literature od 1992. do 2011. godine)

Theobromine and its related compounds, such as caffeine and theophylline, are secondary metabolites that belong to the alkaloids and have economic and cultural importance. These alkaloids have demonstrated stimulatory effects on the central nervous, gastrointestinal, cardiovascular, renal and respiratory systems, resulting in \u27energy arousal\u27, increased motivation to work, increased alertness and increased cognitive function. Several analytical methods have been used to analyse these compounds, but reversed phase high-performance liquid chromatography (RP-HPLC) is the most commonly applied because of its efficiency, sensitivity, specificity and speed. This review describes the analyses of theobromine-related compounds by RP-HPLC with ultraviolet detection (UV) in four sources: food, beverages, biological fluids and plants. Many RP-HPLC methods have been developed and optimized for the detection and quantification of these natural compounds. Elution under isocratic conditions is the most frequent method, with a water, methanol and acetonitrile mixture modified with acetic, phosphoric or formic acid as the mobile phase. For xanthine analysis, the use of reversed phase high-performance liquid chromatography with an ultraviolet/diode array detector (UV/DAD) is particularly suitable as derivation is not required; it allows the analysis of absorbance at all wavelengths, it is simple and rapid.Teobromin i srodni spojevi, kao što su kafein i teofilin, su sekundarni metaboliti koji pripadaju u skupinu alkaloida te imaju ekonomsko i kulturno značenje. Ti alkaloidi djeluju stimulativno na središnji živčani sustav, ali i na probavni, krvožilni, urinarni te dišni sustav, odnosno podižu energiju, motivaciju za rad i poboljšavaju kognitivne funkcije. Za analizu tih spojeva primijenjeno je nekoliko metoda, među kojima se visokodjelotvorna tekućinska kromatografija obrnutih faza (RP-HPLC) pokazala najosjetljivijom, najučinkovitijom i najbržom. U ovom je radu opisana analiza teobromina i srodnih spojeva u hrani, piću, biološkim tekućinama i biljkama pomoću visokodjelotvorne tekućinske kromatografije obrnutih faza s UV-detekcijom. Razvijene su i optimirane različite metode tekućinske kromatografije za analizu prirodnih spojeva. Najčešća je metoda ekstrakcije u izokratskim uvjetima otopinom vode, metanola i acetonitrila modificiranom pomoću mobilne faze (octena, fosforna ili mravlja kiselina). Primjena visokodjelotvorne tekućinske kromatografije s UV/DAD detektorom za analizu ksantina osobito je prikladna jer je jednostavna, brza i omogućuje mjerenje apsorpcije na svim valnim duljinama

Análise fitoquímica e atividades antimicrobiana e antioxidante das folhas de Henriettea succosa (Aubl.) DC

Henriettea succosa is a tree species consumed in abundance by birds, however, there is no report on its phytochemical profile and biological activity. This study performed the phytochemical screening and the antimicrobial and antioxidant potential of H. succosa leaves. The hexane (Hex), ethyl acetate (AcOEt) and methanol (MeOH) extracts of the leaves were evaluated for chemical composition by Thin Layer Chromatography and spectrophotometric analysis; the antimicrobial activity was determined by the Minimum Inhibitory Concentration (MIC) and Minimum Microbicide Concentration (MMC); antioxidant activity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, determination of the reducing power and the phosphomolybdenum complex reduction assay. The photoprotor action of the extracts was also evaluated. The results showed a higher content of phenolic compounds (444.08 ± 0.020 mg EAG/g) and tannins (414.37 ± 0.16 mg EAG/g) in the MeOH extract, which was effective against Staphylococcus aureus and Serratia marcescens, with MIC of 1 mg/ ml and CMM of 2 mg/ml. The MIC and MMC of AcOEt for Micrococcus luteus was 1 mg/mL, this was also considered the minimum concentration necessary for the Hex extract to act on the S. aureus strain. The MeOH extract showed greater antioxidant activity by the DPPH (79.09%) and reducing power (327.2 ± 0.00 mg EAA/g) methods, while the AcOEt extract showed greater activity by the phosphomolybdenum method (40.5%). However, none of the extracts showed a photoprotective effect against UV radiation. In summary, this study revealed that the leaves of H. succosa have secondary metabolites with bactericidal potential, in addition to antioxidant action.Henriettea succosa é uma espécie arbórea consumida em abundância por pássaros, no entanto, não há relatos sobre seu perfil fitoquímico e atividade biológica. Este estudo realizou a triagem fitoquímica e analisou o potencial antimicrobiano e antioxidante das folhas de H. succosa. Os extratos hexano (Hex), acetato de etila (AcOEt) e metanólico (MeOH) foram avaliados quanto a composição química por Cromatografia em Camada Delgada e análise espectrofotométrica; a atividade antimicrobiana foi determinada pela Concentração Inibitória Mínima (CIM) e Concentração Mínima Microbicida (CMM); a atividade antioxidante foi determinada usando-se os ensaios de sequestro do radical 2,2-difenil-1-picrilhidrazil (DPPH), de determinação do poder redutor e de redução do complexo fosfomolibdênio. Foi avaliada também a ação fotoprotetora dos extratos. Os resultados mostraram maior teor de compostos fenólicos (444,08 ± 0,020 mg EAG/g) e taninos (414,37 ± 0,16 mg EAG/g) no extrato MeOH, sendo este eficaz contra Staphylococcus aureus e Serratia marcescens, com CIM de 1 mg/mL e CMM de 2 mg/mL. O CIM e CMM do AcOEt para Micrococcus luteus foi 1 mg/mL, essa também foi considerada a concentração mínima necessária para o extrato Hex agir sobre a cepa de S. aureus. O extrato MeOH apresentou maior atividade antioxidante pelos métodos de DPPH (79,09%) e poder redutor (327,2 ± 0,00 mg EAA/g), enquanto o extrato AcOEt apresentou maior atividade por meio do método fosfomolibdênio (40,5%). No entanto, nenhum dos extratos apresentaram efeito fotoprotetor contra radiação UV.  Em suma, este estudo revelou que as folhas de H. succosa possuem metabólitos secundários com potencial bactericida, além de ação antioxidante

Aplicação de tanase obtida de Aspergillus tamarii URM 7115 e seu efeito em suco de caju integral

Tannin Acyl Hydrolase is an enzyme that catalyzes the hydrolysis of hydrolysable tannins and can be used in several industrial areas, such as food and pharmaceutical. The present work had as objective to produce tannase obtained from Aspergillus tamarii URM 7115 under submerged fermentation and to verify its effect on the integral cashew juice. The total tannins, total phenolics, antioxidant activity and some physical-chemical parameters (pH, °Brix, vitamin C and total acidity) were analyzed before and after application of the enzyme. The tests showed a reduction of at least 50% of the total tannin content, 47-54% increase in phenolics and an increase in total antioxidant activity of 20 to 23%. In addition, the samples of cashew juice before and after enzymatic application presented acceptable results for pH, °Brix, vitamin C and total acidity. The enzymatic treatment in the cashew juice to reduce the astringency obtained several advantages, such as functional quality higher due to the increase in total antioxidant activity, resulting in less turbidity and there was no loss in the nutritional and physical-chemical quality of the juice. Keywords: Anarcadium occidentale L.; Antioxidant activity; Tannin Acyl Hydrolase.Tanino Acil Hidrolase é uma enzima que catalisa a hidrólise de taninos hidrolisáveis e pode ser utilizada em diversas áreas industriais, como alimentícia e farmacêutica. O presente trabalho teve como objetivo produzir tanase obtida de Aspergillustamarii URM 7115 sob fermentação submersa e verificar seu efeito no suco de caju integral. Foram analisados os teores de taninos totais, fenólicos totais, atividade antioxidante e alguns parâmetros físico-químicos (pH, °Brix, vitamina C e acidez total) antes e pós aplicação da enzima. Os ensaios demonstraram uma redução de no mínimo 50 % do teor total de taninos, aumento entre 47 a 54 % no teor de fenólicos totais além de ser verificado um aumento na atividade antioxidante total de 20 a 23 %. Ademais, as amostras do suco de caju integral antes e após aplicação enzimática apresentaram resultados aceitáveis para pH, °Brix, vitamina C e acidez total. O tratamento com a tanase em suco de caju integral, além da redução da adstringência foi observada outras vantagens como aumento da atividade antioxidante, menor turvação e manutenção da qualidade nutricional e físico-química

Production, Characterization and Application of a Thermostable Tannase from Pestalotiopsis guepinii URM 7114

Tanaza (EC 3.1.1.20) je enzim koji hidrolizira esterske veze i razgrađuje taničnu kiselinu na galnu kiselinu i glukozu. Koristi se u proizvodnji hrane i pića za uklanjanje nepoželjenog učinka tanina. Svrha je ovoga rada bila ispitati mogućnost uporabe endofitskih gljivica izoliranih iz lista crne šljive (Syzygium cumini (l.) Skeels), koje pripadaju vrsti Pestalotiopsis guepinii, u proizvodnji tanaze. Tanaza je dobivena submerznim uzgojem, uzgojem u polučvrstoj, te na čvrstoj podlozi. Najviše je tanaze (98,6 U/mL) proizvedeno submerznim uzgojem. Metodom je odzivnih površina ispitan učinak pH-vrijednosti i temperature, a najbolji su uvjeti za aktivnost tanaze bili: pH-vrijednost od 6,9 i temperatura od 30 °C. Vrijednosti su kinetičkih parametara bile sljedeće: Km=7,18·10-4 mol/L i vmax=250,00 U/mL. Aktivnost je tanaze bila najveća u prisutnosti iona Ca2+ koncentracije od 5·10-3 mol/L. Osim toga, ispitan je učinak kelatora i deterdženata, te je utvrđeno da nisu inhibirali aktivnost enzima. Stabilnost je enzima ispitana simulacijom gastrointestinalne probave monogastričnih životinja. Sirovi je enzim bio vrlo stabilan u simuliranim uvjetima, te je nakon šest sati zadržao 87,3 % početne aktivnosti. Ovo istraživanje pridonosi identifikaciji mikrobioloških organizama koji proizvode tanazu, a što bi se moglo primijeniti u biotehnologiji.Tannase (EC 3.1.1.20) is an enzyme that hydrolyzes the ester and depside bonds of tannic acid to gallic acid and glucose. In the production of foods and beverages, it contributes to the removal of the undesirable effects of tannins. The aim of this study is to investigate the potential of endophytic fungi isolated from jamun (Syzygium cumini (L.) Skeels) leaves, and identified as Pestalotiopsis guepinii, in the production of tannase. Tannase was produced extracellularly by P. guepinii under submerged, slurry-state and solid-state fermentations. The submerged fermentation was found to be the most promising (98.6 U/mL). Response surface methodology was employed to evaluate the effect of variables (pH and temperature), and the results showed that the best conditions for tannase activity were pH=6.9 and 30 °C. Km was found to be 7.18·10–4 mol/L and vmax=250.00 U/mL. The tannase activity was the highest in the presence of Ca2+ at a concentration of 5·10–3 mol/L. Moreover, the enzyme was not inhibited by the tested chelators and detergents. The stability of the enzyme was also studied, and crude enzyme was evaluated in simulation of gastrointestinal digestion of monogastric animals. The crude enzyme was highly stable under simulated conditions; it retained 87.3 % of its original activity after 6 h. The study contributes to the identification of microbial species that produce tannase, with potential application in biotechnology

Phytochemical analysis and biological activity from clidemia capitellata leaves (melastomataceae)

This research determined the phytochemical profile and evaluated the photoprotective, antimicrobial and antioxidant activities in extracts of Clidemia capitellata leaves, obtained with hexane, ethyl acetate and methanol. The phytochemical profile was determined through thin layer chromatography; tannins, flavonoids and total phenolics were measured; the antimicrobial activity was determined by Minimum Inhibitory Concentration and Minimum Microbicide Concentration in bacteria Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis and Enterococcus faecalis; Escherichia coli, Serratia marcescens and Pseudomonas aeruginosa, Mycobacterium smegmatis, and the yeast Candida albicans. Antioxidant activity was evaluated through the methods of Reducing Power, DPPH sequestration and phosphomolybdenum. The C. capitellata leaves show flavonoids, tannins, triterpenes, steroids, anthraquinones and essential oils; tannins, flavonoids and total phenolics were extracted in greater amounts with methanol, however, the acetate extract stood out in relation to antimicrobial activity, and none of the extracts showed photoprotective activity within the values established by Brazilian legislation.   Word keys: secondary metabolites. antioxidant. antimicrobial.Esta investigación determinó el perfil fitoquímico y evaluó las actividades fotoprotectoras, antimicrobianas y antioxidantes en extractos de hojas de Clidemia capitellata, obtenidos con hexano, etilo y metanol. El perfil fitoquímico se determinó mediante cromatografía en capa fina; se midieron taninos, flavonoides y fenoles totales; la actividad antimicrobiana se determinó mediante Concentración Mínima Inhibidora y Concentración Mínima de Microbicida en bacterias Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis y Enterococcus faecalis; Escherichia coli, Serratia marcescens y Pseudomonas aeruginosa, Mycobacterium smegmatis y la levadura Candida albicans. La actividad antioxidante se evaluó mediante los métodos de Poder Reductor, secuestro de DPPH y fosfomolibdeno. Las hojas de C. capitellata presentan flavonoides, taninos, triterpenos, esteroides, antraquinonas y aceites esenciales; los taninos, flavonoides y fenoles totales se extrajeron en mayor cantidad con metanol, sin embargo, el extracto se destacó en relación a la actividad antimicrobiana, y ninguno de los extractos mostró actividad fotoprotectora dentro de los valores establecidos por la legislación brasileña. Palabras llaves: metabolitos secundarios. antioxidante. antimicrobiano.Esta pesquisa determinou o perfil fitoquímico e avaliou as atividades fotoprotetora, antimicrobiana e antioxidante de extratos de folhas de Clidemia capitellata, obtidos com hexano, acetato de etila e metanol. Cromatografia em camada delgada foi usada para identificação dos metabólitos; taninos, flavonóides e fenólicos totais foram medidos; a atividade antimicrobiana foi determinada pela Concentração Inibitória Mínima e Concentração Microbicida Mínima nas bactérias Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis e Enterococcus faecalis; Escherichia coli, Serratia marcescens e Pseudomonas aeruginosa, Mycobacterium smegmatis e a levedura Candida albicans. A atividade antioxidante foi avaliada através dos métodos de Poder Redutor, sequestro de DPPH e fosfomolibdênio. As folhas de C. capitellata apresentam flavonóides, taninos, triterpenos, esteróides, antraquinonas e óleos essenciais; taninos, flavonóides e fenólicos totais foram extraídos em maiores quantidades com metanol, porém, o extrato acetato se destacou em relação à atividade antimicrobiana, e nenhum dos extratos apresentou atividade fotoprotetora dentro dos valores estabelecidos pela legislação brasileira. Palavras-chave: metabólitos secundários. antioxidante. antimicrobiano

Analysis of Theobromine and Related Compounds by Reversed Phase High-Performance Liquid Chromatography with Ultraviolet Detection: An Update (1992–2011)

Theobromine and its related compounds, such as caffeine and theophylline, are secondary metabolites that belong to the alkaloids and have economic and cultural importance. These alkaloids have demonstrated stimulatory effects on the central nervous, gastrointestinal, cardiovascular, renal and respiratory systems, resulting in 'energy arousal', increased motivation to work, increased alertness and increased cognitive function. Several analytical methods have been used to analyse these compounds, but reversed phase high-performance liquid chromatography (RP-HPLC) is the most commonly applied because of its efficiency, sensitivity, specificity and speed. This review describes the analyses of theobromine-related compounds by RP-HPLC with ultraviolet detection (UV) in four sources: food, beverages, biological fluids and plants. Many RP-HPLC methods have been developed and optimized for the detection and quantification of these natural compounds. Elution under isocratic conditions is the most frequent method, with a water, methanol and acetonitrile mixture modified with acetic, phosphoric or formic acid as the mobile phase. For xanthine analysis, the use of reversed phase high-performance liquid chromatography with an ultraviolet/diode array detector (UV/DAD) is particularly suitable as derivation is not required; it allows the analysis of absorbance at all wavelengths, it is simple and rapid

Changes in enzymes, phenolic compounds, tannins, and vitamin C in various stages of jambolan (Syzygium cumini Lamark) development

The physiological state of a fruit is closely related to ripening and climatic conditions during the growing period when the fruit undergo changes in color, texture, and flavor. The ripening of the fruit can involve a complex series of biochemical reactions with alteration in enzymes activities, phenols, tannins, and ascorbic acid. The activity of enzymes (carboximethylcellulase, polygalacturonase, and pectinlyase), the total concentration of phenolic compounds, condensed tannins, and vitamin C in five stages of maturation were studied. Significant changes were observed between the maturity stages. The phenolic compounds were higher at green stage (705.01 ± 7.41); tannins were higher at green/purple stage (699.45 ± 0.22). The results showed that the ascorbic acid levels of the pulp varied significantly from 50.81 ± 1.43 to 6.61 ± 1.04 mg.100 g-1 during maturation. The specific activity of pectin lyase was higher at green stage (1531.90 ± 5.83). The specific activity of polygalacturonase was higher at mature stage (1.83 ± 0.0018). The specific activity of carboximetilcelulose was higher at ripe mature stage (4.61 ± 0.0024). The low ascorbic acid content found in jambolan fruit indicates that this fruit is not a rich source of this nutrient; however, other characteristics can make jambolan products fit for human consumption

Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis Aplicação do modelo Doehlert na otimização das variáveis experimentais para a lise de Pseudozyma sp. (CCMB 306) e Pseudozyma sp. (CCMB 300)

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 &#946;-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.<br>O presente trabalho visou verificar a influência do pH e temperatura na lise de leveduras utilizando planejamento experimental. No estudo, foi utilizado o extrato enzimático, contendo &#946;-1,3-glucanase e quitinase líticas, obtidas do micro-organismo Moniliophthora perniciosa. O delineamento experimental mostrou que as melhores condições para a lise de Pseudozyma sp. (CCMB 306) e Pseudozyma sp. (CCMB 300), pelas enzimas líticas, foram pH 4,9 a 37 ºC, pH 3,9 a 26,7 ºC, respectivamente. As enzimas líticas podem ser utilizadas para a obtenção de vários produtos biotecnológicos a partir de leveduras