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As proteases são biocatalisadores com funções fisiológicas complexas, presentes em todos os seres vivos que predominam em processos biotecnológicos nos diversos segmentos da indústria. Das fontes naturais de protease, espécies de Pleurotus sintetizam compostos bioativos de interesse industrial, inclusive enzimas proteolíticas. Neste estudo foi investigada a produção e caracterização parcial de proteases de P. eryngii DPUA 1816 para futura aplicação industrial. A fermentação foi realizada em extrato de Malte ou Sabouraud + extrato de levedura 0,5% (p/v), por sete dias, a 28 ºC, 150 rpm. Nos extratos recuperados foi determinada atividade qualitativa de proteases e avaliado o efeito do pH, temperatura, íons e inibidores na atividade e estabilidade enzimática. Os resultados mostraram que, nas condições experimentais a atividade de proteases de P. eryngii foi 22,70 U/mL, com atividade ótima em pH 5,0, a 40 ºC, com estabilidade na faixa de 40 °C a 80 °C. Os íons Mn2+, Zn2+, Mg2+ e K+ aumentaram a atividade das enzimas em 85%, 84%, 40%, e 31%, respectivamente. Porém, Fe2+ e Cu2+ reduziram ação catalítica em 38% e 37%, respectivamente. O EDTA e o PMSF causaram inibição de 96% e 95,10%, respectivamente. Com base nesses resultados foi observada a presença de metaloproteases e serinoproteases, enzimas que tem importância industrial para elaboração de laticínios e fármacos.

Bee’s pollen comes from the flowers of several species of plants. The grains are transported by worker bees to their nests (honeycombs) to be used as a source of nutrients. The objective of this study was to evaluate the microbiological control, the physical chemical composition and to analyze macro and micro minerals, as well as amino acids in dehydrated pollen hives of Melipona seminigra, in rainy periods (PLL) and dry periods (PS) in Amazonas, Brazil.The physical-chemical composition (proteins, lipids, carbohydrates, fibers and ashes) was performed by the AOAC methodology. In meliponícola pollen it was detected the presence of proteins (35,97 - 36,37 g/100g), lipids (22,37 - 29,66 g/100g), ashes (4,46 - 2,83 g/100g) and ratio carbon/nitrogen (8,94 - 8,28 g/100), also minerals like copper, iron, manganese, calcium, potassium, magnesium and phosphorus were present. In the pollen of the indigenous bees, both non-essential and essential amino acids were found. The results showedthat the meliponícola pollen did not present any contamination for bacteria and fungi that comply with the Brazilian legislation. The physical-chemical analysis confirmed that pollen has nutritional amounts that can be added to other ingredients, providing enrichment of fibers, minerals and proteins to foods at different ages of the population

Alimentos saudáveis com propriedades terapêuticas está impulsionando a produção e a comercialização de espécies de Pleurotus. Em escala comercial, esses cogumelos são cultivados em matriz sólida, utilizando diferentes substratos orgânicos. Resíduos lignocelulósicos disponíveis na Amazônia vêm demonstrando potencial para obtenção de basidiomas de Pleurotus. Este trabalho apresenta contribuições científicas promissoras que demonstram os ciclos de produção de Pleurotus ostreatoroseus e P. albidus, cultivados em resíduos da fruticultura amazônica pela tecnologia da fermentação no estado sólido. Essas espécies desenvolveram a fase micelial completa em grãos de trigo (spwan), em 15 dias e cinco dias, respectivamente. Porém, demonstraram diferentes ciclos produtivos quando cultivadas em exocarpo de cupuaçu misturado com farelo de arroz. P. ostreatoroseus apresentou dois ciclos de produção. No entanto, P. albidus foi observado apenas um fluxo produtivo, caracterizado pela maturação e colheita dos basidiomas. Resíduos lignocelulósicos da fruticultura amazônica são substratos eficientes para a produção de P. ostreatoroseus e P. albidus.

Proteases sao enzimas de elevada importância economica e com ampla aplicabilidade industrial. Enzimas proteoliticas com atividade coagulante do leite bovino, sintetizadas por Aspergillus sp., vem demonstrando caracteristicas bioquimicas com potencial para uso na producao de queijos. O objetivo deste trabalho foi avaliar a producao de proteases por treze culturas de Aspergillus provenientes da colecao de culturas DPUA da Universidade Federal do Amazonas. A avaliacao da viabilidade das especies e pureza das colonias foi realizada com base nas caracteristicas macro, micromorfologicas e forma de reproducao. Apos autenticacao, as especies de Aspergillus foram submetidas ao cultivo em meio liquido para producao de enzimas proteoliticas. A determinacao da atividade proteolitica foi realizada em meio solido e por colorimetria, utilizando como substratos agar leite e solucao de azocaseina 1% (p/v), respectivamente. As proteases do fungo que apresentou atividade proteolitica significativa pela analise colorimetrica foram submetidas a caracterizacao bioquimica. A producao de proteases coagulantes do leite bovino pelas especies de Aspergillus tambem foi determinada. Nos testes de autenticacao foi verificada a viabilidade de 100% das especies de Aspergillus . As proteases das especies de Aspergillus demonstraram diferentes perfis de degradacao do substrato, formando halos translucidos e opacos em meio solido. Os halos de hidrolise significativa foram determinados nos extratos brutos de A . niger DPUA 399 (39,67 mm) e A . japonicus DPUA 1727 (34,27 mm). Na avaliacao da sintese de protease utilizando tecnica colorimetrica foi verificada producao significativa por A. oryzae DPUA 541 (32,00 U/mL). Este fungo sintetiza serinoproteases e metaloproteases que demonstram atividade catalitica otima em pH 5, a 50 °C, mantendo estabilidade na faixa de pH 7 a 10, ate 40 °C. Atividade coagulante significativa foi determinada nos extratos de A. melleus DPUA (19,84 U) e A. oryzae DPUA 541 (19,74 U). Entretanto, A. oryzae DPUA 541 e A. oryzae DPUA 1624 sintetizaram enzimas proteoliticas com forte coagulacao do leite bovino. Os resultados indicam o potencial das especies de Aspergillus avaliadas como forte de proteases com aplicabilidade industrial na area de alimentos, em especial para a producao de queijos.

O objetivo deste estudo foi investigar o efeito do pH, da temperatura e da concentração de nitrato de prata (AgNO3) na síntese de nanopartículas de prata (AgNPs) por Aspergillus flavo-furcatis e avaliar o respectivo potencial antimicrobiano. Um experimento fatorial 2³ foi realizado para avaliar o efeito das condições físico-químicas na síntese de AgNPs. A caracterização das AgNPs foi realizada em espectroscopia UV-Vis e ICP-MS. A atividade antimicrobiana foi avaliada por difusão em ágar por poço e microdiluição em caldo. O pH e a concentração de AgNO3 tiveram efeitos significativos na síntese de AgNPs nas condições experimentais. As AgNPs sintetizadas em pH 9,0 e concentração de AgNO3 de 1,5 mM apresentaram maior eficácia contra Staphylococcus aureus (3,17 µg/mL), Escherichia coli (6,35 µg/mL) e Candida albicans (0,79 µg/mL). Os dados mostraram que a síntese de AgNPs por A. flavo-furcatis é influenciada pelo pH e pela concentração de AgNO3, indicando que as AgNPs sintetizadas possuem atividade antimicrobiana contra S. aureus, E. coli e C. albicans.

Mushrooms have stood out as sources of proteases with industrial applicability. The objective of this work was to evaluate the parameters that influence the synthesis of proteases by Lentinus villosus Klotzsch (1833) AM 169 and to characterize these enzymes. The matrix culture, prepared on potato dextrose agar supplemented with yeast extract, was maintained for 8 days at 25 ° C. Submerged fermentation was carried out in GYPG (glucose, yeast extract, peptone and gelatin). Azocasein solution (1%, w/v) was used as an enzyme substrate. The best conditions for protease production were: initial pH of the culture medium (6), fermentation time (12 days), age of the inoculum (6 days) and size of the inoculum (10%). The proteases were active at pH 8, at 60 ° C, stable at pH (5 to 9) and temperature (30 °C to 60 °C). These enzymes were classified as cysteine proteases, stimulated by Fe 2+ and Mn 2+ . Lentinus villosus synthesizes proteases with potential for application in the detergent, textile, leather processing and bioremediation processes.

Trichosporon yeasts are widely employed to produce lipids, lipases, and aspartic peptidases, but there are no previous studies on collagenase production. This work aimed to select the best collagenase producing Amazonian Trichosporon strains. Moreover, a 2

Proteases são enzimas de origem vegetal, animal e microbianos, de alto valor econômico e biotecnológico, e entre estes, tem destaque as produzidas por espécies de fungos. O presente estudo teve como objetivo analisar a produção de proteases por Aspergillus clavato flavus DPUA 929. A cultura matriz foi preparada em Ágar Sabouraud, com extrato de levedura 0,5% (p/v). O bioprocesso foi conduzido durante 7 dias, 150 rpm a 25 °C. Para determinação da atividade proteolítica, como substrato foi utilizado solução de Azocaseína 1% (p/v). Nas análises de caracterização parcial foi avaliado o efeito do pH e temperatura na atividade das enzimas, conforme protocolo experimental. Os resultados revelaram que Aspergillus clavato flavus DPUA 929 produz proteases equivalente a 21,12 U/mL. E nos testes de caracterização foi determinada a máxima atividade proteolítica em pH 6,0 e a 50 ºC. Estes dados demonstram que Aspergillus clavato flavus, nas condições de análises, produz diferentes proteases com características de importância industrial.

O uso de enzimas no processamento de alimentos, detergente e no setor têxtil está em constante ascensão. A aplicação de enzimas proteolíticas de origem microbiana se destaca no setor industrial devido as condições desejáveis para uso em processos biotecnológicos, são biocatalisadores naturais e não tóxicos. O objetivo dessa pesquisa foi avaliar a produção e caracterizar as proteases sintetizadas por Aspergillus flavus. A cultura matriz foi preparada em ágar Sabouraud e mantida por sete dias a 25 ºC. Em seguida, a espécie foi cultivada em meio líquido [GYP (glicose, extrato de levedura e peptona)]. A fermentação foi conduzida em agitador orbital 150 rpm, 25 ºC. Após oito dias a massa micelial foi separada por filtração a vácuo em papel Whatman nº 1 e o extrato bruto recuperado, e filtrado em membrana polietersulfônica de 0,45μL. Para determinação da atividade das proteases foi utilizado como substrato azocaseína 1% (p/v) em tampão Tris-HCl pH 7.2. Os resultados mostraram que A. flavus excretou proteases em todos nos meios testados, porém com atividade significativa no meio GYP (90,13 U/mL). As proteases apresentaram atividade ótima em pH 5 e temperatura 50ºC. Esses resultados sugerem que essas enzimas podem ser utilizadas em diversos segmentos industriais, como alimentícia, têxtil e panificação.

Silver nanoparticles (AgNP) obtained from biological synthesis can be widely used in industrial and medical fields because of their observed antimicrobial activity. The objective of this study was to analyze the biosynthesis of AgNPs by the fungus Lentinus crinitus (L.) Fr., and to evaluate the potential of these nanoparticles as antimicrobial agents. The antimicrobial activity of AgNPs was evaluated by agar diffusion, and broth microdilution methods to determine the minimum inhibitory concentration (CMI) against Staphylococcus aureus, Escherichia coli, Candida albicans and Candida tropicalis. AgNPs were characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), inductively coupled plasma emission spectrometry (ICP) and transmission electron microscopy (TEM). The UV-Vis spectra of the reaction mixture showed a SPR band with peak absorbance at 423 nm, confirming the presence of AgNPs. The synthesized AgNPs demonstrated antagonistic action against C. tropicalis (1.88 µg. mL-1), C. albicans (30.09 µg. mL-1), E. coli and S. aureus (7.52 µg. mL- 1). The AgNPs mediated by L. crinitus are mostly spherical, triangular and rod-shaped (mean diameter 8.82 nm). The concentration of silver in their crystalline structure is 120.37 µg / mL, and protein residues as possible stabilizers. The Lentinus crinitus mushroom isolated from substrates of the Amazon biome is a promising bio-resource for the biological synthesis of AgNPs with relevant antimicrobial properties and demonstrating a great potential for its application in pharmaceutical and food industries.

As proteases têm elevada aplicação industrial e, nos últimos anos, os cogumelos têm se destacado como fontes naturais e renováveis destes biocatalisadores. O objetivo desta pesquisa foi avaliar a produção e caracterizar as proteases sintetizadas por Lentinus crinitus (L.) Fr. 1825 DPUA 1693. A cultura matriz, preparada em ágar batata dextrose, suplementado com 0,5% (p/p) de extrato de levedura, foi mantida por 8 dias a 25 °C. Em seguida, L. crinitus foi cultivado em uma mistura de farelo de arroz e semente de tucumã durante 15 dias a 25 °C. As proteases foram extraídas em água destilada esterilizada e, no extrato bruto, foi determinada a atividade proteolítica, utilizando-se como substrato azocaseína 1 % (p/v). As condições que favoreceram a síntese destas enzimas foram: pH inicial do meio de cultivo (8,0), tempo de fermentação (12 dias), idade do inóculo (seis dias) e tamanho do inóculo (10%). As proteases demonstraram as seguintes características bioquímicas: pH ótimo (6,0), temperatura ótima (50 °C), estabilidade na faixa de pH (5,0-6,0) e temperatura (30-50 °C). As proteases foram estimuladas por íons Mn2+ e classificadas como aspártico proteases. Estes biocatalisadores têm potencial para produção de queijos, pães e bebidas.

Considerando-se o desenvolvimento de produtos utilizando matéria-prima regional, foram extraídos e caracterizados os amidos de Dioscorea alata L. e D. altissima Lam. Os parâmetros analisados foram rendimento, composição, morfologia e tamanho dos grânulos, composição físico-química, propriedades tecnológicas, toxicidade in vitro e análise microbiológica. Rendimento significativo (8,57%) foi observado em D. altissima. Amido disponível (75,02%), total (84,63%) e amilose (19,15%) foram maiores em D. alata. Amido resistente (10,10%) e amilopectina (82,09%) foram superiores em D. altissima. O pH e a acidez não variaram entre as amostras. A atividade de água foi menor em D. alata. Baixos teores de umidade, cinzas, lipídios, proteínas, fibras e alto conteúdo de carboidratos foram determinados. Os amidos não apresentaram citotoxicidade e nem contaminação microbiana. Os grânulos do amido de D. altissima são esféricos e significativamente maiores. Nos amidos de D. alata e de D. altissima, predominam, respectivamente, macro e microminerais. Absorção de água (85,73 g 100g-1), óleo (67,13 g 100g-1), estabilidade da espuma (100%) e claridade da pasta (2,87%) foram maiores em D. alata. A capacidade de inchamento (4,07 g g-1) e a densidade aparente (0,87g mL-1) foram superiores em D. altissima. Os amidos estudados possuem propriedades tecnológicas com potencial para a indústria de alimentos e para a produção de embalagens.

Proteases são enzimas hidrolíticas capazes de degradar proteínas em pequenos peptídeos e aminoácidos, e podem ser isoladas de plantas, animais e microrganismos. A busca por proteases de origem microbiana vem aumentando no mercado mundial devido à sua estabilidade, à alta taxa de produção e ao menor custo que apresenta. O objetivo deste artigo foi investigar a produção de proteases por Lentinus crinitus (L.) Fr. 1825 DPUA 1693, utilizando-se a fermentação submersa. A cultura matriz foi preparada em meio BDA (ágar batata dextrose), adicionando-se extrato de levedura 0,5% (p/v). A fermentação foi conduzida a 30 °C, 150 rpm, por oito dias, utilizando-se meios contendo diferentes fontes de carbono e nitrogênio na síntese de proteases. Na determinação da atividade enzimática, foi adotada azocaseína 1% (p/v) como substrato. Os resultados mostraram que a maior atividade das proteases de L. crinitus foi determinada no meio contendo glicose e gelatina. A atividade ótima dessas enzimas foi determinada em pH 5,0, a 50 °C, e a estabilidade, em pH 5,0, à temperatura de 60 °C. As proteases foram classificadas como metalo, serino e cisteíno proteases. Este artigo sugere o uso potencial de protease de Lentinus crinitus para aplicação na indústria alimentícia, farmacêutica e cosmética.

Made available in DSpace on 2019-10-06T15:48:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-08-16 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The replacement of synthetic colors in food products by natural alternatives has been boosted by consumers willing to pay more for healthier products. However, the success of microbial colorants depends not only on its acceptability on the market but also its production costs. Talaromyces species can produce water-soluble red colorants induced by glucose and monosodium glutamate (MSG). In this study, the influence of several conditions was evaluated to produce natural red colorants by submerged culture of Talaromyces amestolkiae. Under optimal conditions (g/L: glucose 10, MSG 25, MgSO4 0.012, FeSO4 0.01, CaCl2 0.015; and initial pH of 5.0), a 30-fold increase in the production was achieved, reaching a red colorant production of 13.44 UA500nm. Depending on the initial pH, colorants with different hues and chroma values were obtained. Deep yellow colorants were derived from neutral and basic pH, while deep red colors were derived from acidic pH. The fluorescence spectrum of culture broth obtained before and after complexation with salts presented red colorants with yellow fluorescence spectra. The information generated in this study would be useful for the formulation of industrial media for large-scale cultivation of T. amestolkiae, which have the potential to produce Talaromyces fermented colorants for use in health foods and pharmaceutics. Department of Bioprocess and Biotechnology School of Pharmaceutical Sciences Campus (Araraquara) Universidade Estadual Paulista – UNESP, Rodovia Araraquara-Jaú/ Km 01, Campos Ville - Araraquara/SP Culture Collection DPUA/UFAM Universidade Federal do Amazonas Department of Bioprocess and Biotechnology School of Pharmaceutical Sciences Campus (Araraquara) Universidade Estadual Paulista – UNESP, Rodovia Araraquara-Jaú/ Km 01, Campos Ville - Araraquara/SP CAPES: 001 CNPq: 155317/2016-4 FAPESP: 2014/01580-3

Fibrinolytic enzymes have been considered promising for treatment and protection of healthy circulation due its ability to dissolve the fibrin in blood clots. Extractive fermentation is a not explored and efficient downstream process which segregates the desired product simultaneously in a fermentation process fast and economically. Extraction of fibrinolytic enzymes by

The objective of this study was to analyze mycelial growth under different culture conditions and antimicrobial activity of Pleurotus ostreatus (Jacq.: Fr.) Kumm. (DPUA 1533) and P. ostreatus (Jacq.: Fr.) Kumm. cv. Florida (DPUA 1534) against fungi and bacteria of medical importance. The growth of Pleurotus species was evaluated in natural and complex media, with and without light, at 25°C and 28°C for 8, 15, and 30 days. Candida albicans (DPUA 1336), Cryptococcus laurentii (DPUA 1501), Aspergillus flavus (DPUA 1836), Escherichia coli (DAUPE 224), and Mycobacterium smegmatis (ATCC 607) were used to test antibiosis. Under all growth conditions in vitro, Pleurotus species evidenced growth and high density of mycelia on potato dextrose agar and Sabouraud agar with yeast extract; mycelial growth but lesser mycelial density was observed on rice bran extract agar. Organic mycelial extracts of Pleurotus species exhibited potential antibacterial and antifungal activity, and were selective for the tested microorganisms.

Pleurotus albidus, a naturally growing species in the Amazon region, has been considered a promising source of milk-clotting proteases. The production of such enzymes using lignocellulosic residues is a sustainable alternative to replace mammalian rennet. The application of P. albidus milk-clotting proteases in cheese making has not yet been reported in the scientific literature. The aim of this study was to characterize the milk-clotting proteases of P. albidus and use these enzymes in the production of Minas frescal cheese. For the production of coagulating proteases, the mushroom was grown in açaí seeds supplemented with rice bran (10%, w/w). The parameters affecting the production of coagulant, such as inoculum size, fermentation time, initial pH of cultivation medium and age of the inoculum were evaluated. The coagulant extract obtained under optimal production conditions was evaluated for optimal pH and temperature, pH and temperature stability, effect of ions and inhibitors. Significant production of coagulating proteases was obtained under the following conditions: inoculum size (2.5%), fermentation time (10 days), initial pH of the cultivation medium (6), and inoculum age (10 days). The coagulant exhibited significant catalytic activity in pH 5.0 at 55°C, with stability at 45°C and was completely inhibited by iodoacetic acid. The milk-clotting proteases of P. albidus were efficient for making Minas frescal cheese that presented 55.0% of moisture, 20.0% of lipids and 17.20% of protein. Pleurotus albidus is a potential source of milk-clotting proteases that can be applied in dairy industry for production of fresh Minas frescal cheese. Pleurotus albidus, a naturally growing species in the Amazon region, has been considered a promising source of milk-clotting proteases. The production of such enzymes using lignocellulosic residues is a sustainable alternative to replace mammalian rennet. The application of P. albidus milk-clotting proteases in cheese making has not yet been reported in the scientific literature. The aim of this study was to characterize the milk-clotting proteases of P. albidus and use these enzymes in the production of Minas frescal cheese. For the production of coagulating proteases, the mushroom was grown in açaí seeds supplemented with rice bran (10%, w/w). The parameters affecting the production of coagulant, such as inoculum size, fermentation time, initial pH of cultivation medium and age of the inoculum were evaluated. The coagulant extract obtained under optimal production conditions was evaluated for optimal pH and temperature, pH and temperature stability, effect of ions and inhibitors. Significant production of coagulating proteases was obtained under the following conditions: inoculum size (2.5%), fermentation time (10 days), initial pH of the cultivation medium (6), and inoculum age (10 days). The coagulant exhibited significant catalytic activity in pH 5.0 at 55°C, with stability at 45°C and was completely inhibited by iodoacetic acid. The milk-clotting proteases of P. albidus were efficient for making Minas frescal cheese that presented 55.0% of moisture, 20.0% of lipids and 17.20% of protein. Pleurotus albidus is a potential source of milk-clotting proteases that can be applied in dairy industry for production of fresh Minas frescal cheese.

As proteases constituem um grupo de enzimas que apresentam elevada aplicabilidade em diversos setores industriais, sendo obtidas de fontes animais, vegetais e microbianas. Entre os fungos, os cogumelos têm se destacado como fonte de proteases com potencial biotecnológico. O objetivo deste trabalho foi investigar a produção de proteases por cogumelos, utilizando a fermentação em meio líquido. A cultura matriz foi preparada em ágar batata dextrose, acrescida de extrato de levedura 0,5% (p/v), e o bioprocesso foi conduzido durante cinco dias, a 150 rpm e 25 °C. Azocaseína 1% (p/v) foi utilizada como substrato para avaliar a atividade proteolítica dos cogumelos. Entre as espécies avaliadas, Pleurotus albidus (Berk.) Pegler 1983 foi a produtora significativa de proteases (34,00 U/mL) e a síntese dessas enzimas foi estimulada pela idade e pelo volume do inóculo. Essas enzimas mostraram atividade ótima em pH 5 a 60 °C e estabilidade em pH 5-8, com temperaturas de 30 a 60 °C. As proteases foram classificadas como cisteíno e serinoproteases.

Proteases are important commercial enzymes, and among their numerous sources are the Basidiomycetes. The use of proteases in many industrial areas promotes the search for enzymes with new properties. The aim of this study is to produce and characterize peptidases of a biocomposite from mycelial biomass grown in Amazonic tubers. Lentinus citrinus DPUA 1535 and Pleurotus ostreatoroseus DPUA 1720 were cultivated on Dioscorea trifida, Manihot esculenta and Dioscorea alata supplemented with rice bran or manioc flour residue in different proportions. The highest proteolytic activity was determined in the crude extract from P. ostreatoroseus grown in D. alata (DA) without supplementation (142.22 U/mL). The enzymes showed optimum activities at 40°C and pH 7.0; and stability at 50°C and pH 8.0. The proteases were classified as cysteine proteases based on the effect of inhibitors used. Key words: Pleurotus, Lentinus, Dioscorea, protease.

The microbial sources used for the production of nanoparticles are important, and one of the reasons for bacterial preference is their ease of manipulation and ecofriendly potential. Three bacteria strains of Streptomyces genus were used for the biosynthesis of silver nanoparticles (AgNPs). The aim of the present study was to confirm Streptomyces spp. as biological systems in the synthesis and extracellular characterization of silver nanoparticles with antimicrobial activity. The nanoparticles were characterized by ultraviolet–visible spectroscopy (UV-Vis), dynamic light scattering (DLS), RDX, Fourier-transform infrared spectroscopy (FTIR) and transmission electron microscope (TEM). In qualitative antimicrobial test, Streptomyces parvulus DPUA 1549 and Streptomyces owasiensis DPUA 1748 showed activity against pathogenic microorganisms (bacteria and yeasts) tested. The AgNPs of S. parvulus DPUA 1549 showed the most efficient minimum inhibitory concentration (MIC) at low concentrations (1.95 μM) against Staphylococcus aureus. They promoted structural damage to the micro-organism cell surface according to AFM images and also reduced the proliferation of NIH-3T3 cells in the cytotoxic assay. Thus, biosynthesis of silver nanoparticles by Streptomyces spp. is a potential source to be used in nanomedicine and cosmetology industries. Key words: Streptomyces, tropical region, nanobiotechnology, antimicrobial, cytotoxic test.

Com os objetivos de verificar a influência do método de preservação na atividade de metabólitos secundários e de selecionar espécies de Aspergillus como fontes de compostos com atividade antimicrobiana, foi feito um estudo com Aspergillus da coleção de culturas do Departamento de Parasitologia da Universidade Federal do Amazonas (DPUA/UFAM). As linhagens foram reativadas e a sua viabilidade foi confirmada com base nas características morfológicas e nas estruturas de reprodução. A produção dos biocompostos foi feita em ágar extrato de levedura sacarose (yeast extrat sucrose – YES). Os extratos orgânicos etanólico, acetato de etila e hexânico foram testados pelo método de difusão em ágar por poço contra Staphylococcus aureus, Escherichia coli e Candida albicans. Os extratos que apresentaram antibiose neste teste foram analisados por bioautografia de imersão, sendo determinadas as concentrações mínimas inibitórias pelo método de microdiluição em caldo. O método de preservação em água destilada promoveu maiores estabilidade e pureza das culturas dos Aspergillus, independentemente do tempo de armazenamento, em comparação ao método de conservação em óleo mineral, sobretudo em relação ao fenômeno do pleomorfismo. Os dados aqui apresentados mostraram que, em condições in vitro, espécies de Aspergillus sintetizam compostos com atividade antibacteriana e antifúngica para inibir o crescimento de S. aureus, E. coli e C. albicans, preservados em água destilada, mesmo que por diferentes períodos.

Silver nanoparticles (AgNPs) are nanoforms that express higher antimicrobial potential due to their shape and reduced size. The use of fungi to mediate the synthesis of AgNPs has increased the interest of scientists because of their rapid growth, large-scale cultivation and secretion of non-toxic molecules. The aim of this study was to synthesize AgNPs mediated by Aspergillus oryzae DPUA 1624 and evaluate the antimicrobial activity of these molecules incorporated in cellulose acetate nanomembranes (NanoMAC). The synthesis of AgNPs was confirmed by UV-visible spectroscopy and the characterization was performed by dynamic light scattering, transmission electron microscopy, X-ray diffraction, X-ray dispersive energy and advanced spectroscopy and spectroscopy methods. The synthesis of the membrane was done by electro-spinning and its thickness was analyzed in scanning electron microscope. The AgNPs were added to NanoMAC and the antimicrobial effect was evaluated by agar diffusion method against Staphylococcus aureus, Escherichia coli and Candida albicans. The aqueous extract of A. oryzae mediated the synthesis of AgNPs with rounded and some triangular shapes. The diameter and zeta potential of these particles were 61.15±11.45 nm and -28.7 mv, respectively. The NanoMAC with AgNPs showed an increase in antifungal activity of 24.22% when tested against C. albicans. This study demonstrated that A. oryzae is able to mediate AgNP synthesis with anti-yeast action and the impregnation of AgNPs in acetate cellulose nanomembranes resulted in the development of a more efficient antimicrobial nanocomposite. Key words: Silver nanoparticles, Aspergillus oryzae, cellulose nanofibers, anti-yeast.

Proteolytic enzymes are metabolites that can be produced by microbial sources and develop important functions in food industry as in cheese manufacturing. However, it is necessary to ensure the final product safety by testing toxins production by microorganisms. As a result of this, the aim of this study was to investigate the production of proteases by Aspergillus flavo furcatis DPUA 1608 in submerged and solid state fermentation and also certify the non-production of aflatoxin by this species. The aflatoxin test was carried out using the method of ammonia vapor. In this test, A. flavo furcatis DPUA 1608 was inoculated in seven different media cultures (COA, YES, CZ, CYA, GMS, PMS and PDA) and the production of toxins was confirmed by the color change of culture reverse after adding a 25% (w/v) ammonia solution. The protease production was conducted using four inoculums (SAB+GLI, SAB+SAC, BDA+GLI and BDA+SAC) in three media cultures of submerged fermentation obtained by a base mineral solution (MA01, MAGli and MASac) and in solid state fermentation using acai seeds and rice bran as substrate (SAFA). According to ammonia vapor test, A. flavo furcatis is not an aflatoxin producer. There was no color change in the colonies reverse of any culture media. All crude extracts obtained in both fermentations were tested for protease production. The best protease activity was observed in the medium MA01 (inoculums SAB+SAC, BDA+GLI and BDA+SAC). Milk clotting activity was determined in all crude extracts of submerged and solid fermentation. However, the clot formed was considered as strong milk coagulation only in the media MA01 of submerged fermentation (inoculums SAB+SAC, BDA+GLI and BDA+SAC) and in all inoculums of solid state fermentation (SAFA). A. flavo furcatis DPUA 1608 showed potential milk-clotting protease production in both fermentations media used. Key words: Ammonia vapor test, Aspergillus, milk-clotting enzymes, liquid fermentation, solid fermentation.

Films and coatings based on natural polymers are used to conserve nutritional quality of fruits, vegetables and also delay their ripening. The purpose of this study was to develop films with starch extracted from Dioscorea altissima Lam. (dunguey) incorporated with silver nanoparticles for coating and preserving fruits. The films obtained by cast were characterized visually, by Scanning Electron Microscope, by Atomic Force Microscopy, by X-Ray Diffraction and by Fourier-Transform Infrared Spectroscopy. Antimicrobial activity and technological properties were also evaluated. The coating of camu-camu fruits [Myrciaria dubia (Kunth) McVaugh] was carried out by immersing them in the filmogenic solution, followed by their physicochemical and microbiological analysis. The films with silver nanoparticles showed transparency, flexibility, spherical clusters and a higher average roughness. A reduction in thickness, solubility and water vapor permeability was also observed. Antimicrobial action against Staphylococcus aureus and Escherichia coli was proved as well. The fruits coated with films exhibited delay in ripening, with maintenance of quality and longevity. Uncoated fruits showed greater wilting and wrinkling. The starch film incorporated with silver nanoparticles was effective for preserving camu-camu fruit. Películas y recubrimientos a base de polímeros naturales son utilizados para conservar la calidad nutricional de frutas, verduras y también para retrasar su maduración. El propósito de este estudio fue desarrollar películas con almidón extraído de la Dioscorea altissima Lam. (ñame) incorporado con nanopartículas de plata para el recubrimiento y conservación de frutas. Las películas obtenidas por fusión se caracterizaron visualmente, por medio de Microscopio Electrónico de Barrido, mediante Microscopía de Fuerza Atómica, con Difracción de Rayos X y por Espectroscopía Infrarroja con Transformada de Fourier. Actividad antimicrobiana y propiedades tecnológicas también fueron evaluadas. El recubrimiento de frutos de camu-camu [Myrciaria dubia (Kunth) McVaugh] se realizó sumergiéndolos en la solución filmógena, seguido por el análisis fisicoquímico y microbiológico. Las películas con nanopartículas de plata mostraron transparencia, flexibilidad, acumulaciones esféricas y un mayor promedio de rugosidad. También se observó una reducción en el espesor, la solubilidad y la permeabilidad al vapor de agua. Asimismo, acción antimicrobiana contra Staphylococcus aureus y Escherichia coli fue evidenciada. Los frutos recubiertos con películas exhibieron retraso en la maduración, con mantenimiento de calidad y longevidad. Los frutos sin recubrir mostraron mayor marchitez y arrugas. La película de almidón incorporada con nanopartículas de plata fue eficaz para preservar a la fruta de camu-camu. Filmes e revestimentos à base de polímeros naturais são usados para conservar a qualidade nutricional de frutas, vegetais e também retardar seu amadurecimento. O objetivo deste trabalho foi desenvolver filmes com amido extraído de Dioscorea altissima Lam. (cará-de-espinho) incorporado com nanopartículas de prata para revestimento e preservação de frutas. Os filmes obtidos por casting foram caracterizados visualmente, por Microscópio Eletrônico de Varredura, Microscopia de Força Atômica, Difração de Raios-X e Espectroscopia de Infravermelho com Transformada de Fourier. A atividade antimicrobiana e as propriedades tecnológicas também foram avaliadas. O revestimento dos frutos de camu-camu [Myrciaria dubia (Kunth) McVaugh] foi realizado por imersão em solução filmogênica, seguida de suas análises físico-químicas e microbiológicas. Os filmes com nanopartículas de prata apresentaram transparência, flexibilidade, aglomerados esféricos e maior rugosidade média. Uma redução na espessura, solubilidade e permeabilidade ao vapor de água também foi observada. A ação antimicrobiana contra Staphylococcus aureus e Escherichia coli também foi comprovada. Os frutos revestidos com filmes exibiram atraso no amadurecimento, com manutenção da qualidade e longevidade. Frutos não revestidos apresentaram maior murchamento e enrugamento. O filme de amido incorporado com nanopartículas de prata foi eficaz na preservação do fruto do camu-camu.

Resíduos hortícolas podem ser utilizados para a cultivo de cogumelos e obtenção de proteases. O objetivo desta pesquisa foi avaliar o potencial de resíduos lignocelulósicos como substratos para o crescimento micelial e produção de proteases por Lentinus crinitus DPUA 1693. A cultura matriz foi preparada em ágar batata dextrose (BDA) suplementada com extrato de levedura (YE) 0,5% (p/v). O crescimento micelial vertical (VMC) foi realizado em tubos de ensaio contendo os seguintes resíduos lignocelulósicos suplementados com farelo de arroz (FA): serragem, (SER), semente de açaí (SA) e exocarpos de abacaxi (CAB), cupuaçu (CC) e tucumã (CT). O vigor micelial foi classificado pelo método subjetivo de notas em: Fracamente adensado (I), Mediamente adensado (II) e Fortemente adensado (III). A atividade proteolítica foi determinada utilizando solução de azocaseína 1% (p/v). A caracterização enzimática foi realizada no extrato obtido do cultivo de L. crinitus no resíduo lignocelulósico que favoreceu o desenvolvimento de micélio fortemente adensado e produção significativa de proteases. A VMC significativa foi determinada em SER+FA (0,40 cm/dia). Em CC+FA, este cogumelo cresceu e formou micélio fortemente adensado (III). Atividade proteolítica significativa foi determinada nos cultivos de L. crinitus em CC+FA (6,71 U/mL) e CT+FA (6,44 U/mL). No extrato bruto de L. crinitus cultivado em CC+FA houve predomínio de aspártico proteases que demonstraram atividade ótima em pH 5, a 50 °C. Essas enzimas foram fortemente inibidas por Zn+2 e Mn+2. Os resíduos lignocelulósicos utilizados favorecem a produção de massa micelial e proteases por L. crinitus.

Cogumelos do gênero Pleurotus são cultivados para fins alimentícios, medicinais e por serem fontes de enzimas de interesse industrial. O presente estudo teve como objetivo avaliar o crescimento micelial e verificar a atividade de proteases de P. djamor. Este cogumelo foi cultivado em ágar batata dextrose (BDA) com extrato de levedura (YE) 0,5% (p/v). Para avaliar a velocidade do crescimento micelial radial (VCR) e densidade micelial, P. djamor foi cultivado em cinco meios com YE 0,5% (p/v): ágar batata dextrose (BDA+YE), ágar Sabouraud (SAB+YE), ágar extrato de malte (MEA+YE), ágar inhame-roxo (PYA+YE) e ágar cará-de-espinho (SYA+YE). A atividade proteolítica foi determinada pelo método de bloco de gelose. Na avaliação do crescimento micelial vertical (VCM) foram utilizados exocarpos de cupuaçu (CE) e abacaxi (PE), semente de açaí (AS) e serragem (SAW), suplementados com farelo de arroz (RB) ou farelo de trigo (WB). A VCR significativa foi observada em SYA+YE (11,26 mm/dia) e PYA+YE (10,97 mm/dia). Porém, SAB+YE favoreceu a densidade micelial, portanto foi o meio selecionado para obtenção de inóculo nos testes de avaliação do crescimento micelial de P. djamor em resíduos lignocelulósicos. Atividade proteolítica significativa foi determinada em BDA+YE (17,7 mm), MEA+YE (17,6 mm) e SAB+YE (17,0 mm). Forte densidade micelial e VCM significativa foi determinada em CE+WB (0,37 cm/dia) e CE+RB (0,30 cm/dia). Substratos de floresta tropical são eficientes para o cultivo micelial e produção de proteases por P. djamor.

Made available in DSpace on 2021-06-25T10:14:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-02-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Recent environmental and health concerns have been pushing the society for the consumption of natural and sustainable products, as for example, the use of natural colorants as additives to impart color. Talaromyces amestolkiae are natural producers of red colorants at both extracellular and cell-bond state, so are very important for the dye industry. T. amestolkiae cells are constituted by a complex and rigid cell wall structure, which contain a major fraction of the total colorants content. The use of environmentally friendly, biocompatible and high-efficiency cell disruption procedures is thus critical for the recovery of the intracellular colorants. An alternative approach using imidazolium-based ionic liquids (ILs) was applied to recover the intracellular colorants from T. amestolkiae fungi. First, the cell disruption ability of a series 1-alkyl-3-methylimidazolium chloride ILs ([Cnmim]Cl) aqueous solutions was determined, assessing both the effect of increasing cationic alkyl chain length (n = 1, 2, 4, 6, 8 or 10) and IL concentration (30% and 45% w/v). [C4mim]Cl showed the highest extraction ability to recover red, orange and yellow colorants. In the second part, the extraction ability of [C4mim]Cl (at 45% w/v in water) was improved, by combining the IL solution chemical treatment with a mechanical (ultrasound-temperature-assisted) cell disrupting method. At this point, the incubation time (at 60 °C and 150 rpm) after sonication, number sonication pulses (5 min ON) and re-extraction effects were screened as parameters for increasing the cell disruption. The mechanical-assisted method allowed an increase of almost 3-fold in the extractive performance of [C4mim]Cl solution, achieving a maximum recovery of 76.07 UA490nm/mL of red colorants. The results demonstrate the potential of [C4mim]Cl solution as chemical cell-disrupting aid in ultrasound-temperature-based methods to improve the recovery of intracellular colorants from complex filamentous fungi cells. Department of Engineering Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP) Culture Collection DPUA/UFAM Universidade Federal do Amazonas Univ Coimbra CIEPQPF Department of Chemical Engineering Rua Sílvio Lima, Polo II - Pinhal de Marrocos Department of Engineering Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP) FAPESP: 2014/16424-7 FAPESP: FAPESP 2014/01580-3

The aim was to investigate the influence of D. trifida , C. allouia and D. altissima Amazonian tubers in gut bacteria growth in vitro and in vivo . The best result in vitro was D. trifida fiber that stimulated bacterial growth without significative difference of commercial inulin. Because of the great results, D. trifida and C. allouia were chosen for flours production and in vivo tests. These tubers studied were efficient for producing flours source of carbohydrate, tryptophan and valine and minerals both with different prebiotic effects. D. trifida and C. allouia flours have high carbohydrate content (84% and 81%), lower levels of lipids (0.9% and 0.9%), protein (2.3% and 4.2%), fiber (2.2% and 1.9%), minerals and essential amino acids. The rats gut microbiota were influenced by diet with D. trifida demonstrating an expressive bifidogenic effect and C. allouia influenced Bifidobacterium and Lactobacillus population in the animals gut. Industrial relevance The flours of Dioscorea trifida and Calathea allouia are considered a simple technology that guarantees a longer shelf life for those tubers. The modulation effect on gut microbiota of rats is an unpublished evidence that increases value of the product. These results also suggest that the tubers consumption could be stimulated. In this study, we highlight evidences of beneficial health function of these tubers. This scientific study suggests that these tubers flours could be a tool for nutritional therapy. It also incentive the produce of flours to fomenting the productive chain, since the flour tuber technology use can contribute to the local economy.

Resumo: A mastite é uma inflamação na glândula mamária que pode acarretar perdas na produção e na qualidade do leite, gerando prejuízos econômicos para a pecuária leiteira. O tratamento é baseado na utilização de antibióticos, sendo, em muitos casos, ineficazes devido à resistência bacteriana já conhecida para esta doença. O objetivo deste trabalho foi selecionar linhagens de Streptomyces spp. produtoras de biocompostos com atividade antimicrobiana frente a isolados do gênero Staphylococcus multirresistentes de búfalas com mastite. Bem como, determinar os melhores parâmetros de produção, e avaliar a produção simultânea de ácido clavulânico. A seleção de Streptomyces spp. com capacidade de produzir compostos com atividade antimicrobiana foi realizada através da técnica bloco de gelose. Dentre as 30 espécies de Streptomyces spp. testadas, o micro-organismo Streptomyces parvulus DPUA 1573 apresentou melhores resultados, sendo capaz de inibir o crescimento de 7 isolados Staphylococcus spp. multirresistentes. Posteriormente, a espécie selecionada Streptomyces parvulus DPUA 1573 foi cultivada em diferentes condições pré-determinadas pelo planejamento fatorial 24, onde as variáveis independentes foram: concentração de soja (0,5; 1,0; 1,5%), glicose (0; 0,5; 1g/L), agitação (150; 200; 250rpm) e temperatura (28; 32; 37°C) e todos os ensaios do planejamento foram monitorados até 120 horas de cultivo. Todas as variáveis independentes influenciaram positivamente no crescimento celular, enquanto que para atividade antimicrobiana apenas as variáveis temperatura e agitação apresentaram efeitos significativos positivos. O líquido metabólito produzido por Streptomyces parvulus DPUA 1573 foi capaz de inibir o crescimento de sete Staphylococcus spp. multirresistentes. As melhores condições de cultivo para a produção de moléculas bioativas por este micro-organismo foi a 37?C, com 250rpm de agitação por período de 72 horas. Nos ensaios que apresentaram atividade antimicrobiana, foi avaliada a produção de ácido clavulânico ao longo do cultivo. A maior concentração de ácido clavulânico foi de 269,84g/L obtidas nas condições de 1,5% de farinha de soja em ausência de glicose no tempo de 96 horas. A linhagem Streptomyces parvulus DPUA 1573 foi eficiente contra Staphylococcus spp. multirresistentes isolados de mastite em búfalas, ainda apresentando concomitantemente produção de ácido clavulânico com o potencial uso farmacêutico. Abstract: Mastitis is an inflammation in one or more mammary glands which can lead to reduction in production and quality of milk causing economic losses to dairy farming. The use of antibiotics is the key for the treatment of this disease, but in many cases ineffective due to bacterial resistance already known for this condition. The aim of this study was to select strains of Streptomyces spp. producing biomolecules with antimicrobial activity against multidrug-resistant Staphylococcus isolated from buffaloes with mastitis, as well as to determine the best production parameters to the evaluation of simultaneous production of clavulanic acid. Thirty species of Streptomyces spp. were used to selecting the greatest producer spectrum of antimicrobial activity (agar block technique), with selection of Streptomyces parvulus DPUA 1573, and 7 multidrug-resistant Staphylococcus spp. sensitive to its biocompounds. The selected strain of Streptomyces parvulus DPUA 1573 was cultured in different conditions predetermined by the factorial design 24, where the independent variables were: soybean concentration (0.5, 1.0, 1.5%), glucose (0, 0.5, 1g/L), agitation (150, 200, 250rpm) and temperature (28, 32, 37°C); all the tests were monitored up to 120 hours of cultivation. All independent variables influenced positively the cell growth, while for antimicrobial activity only the variables temperature and agitation showed positive effects. The antimicrobial bio compounds showed activity against seven multidrug-resistant Staphylococcus spp under the conditions: temperature 37°C, agitation 250rpm, with 72 hours of production process. In the tests which showed antimicrobial activity, was also assessed the production of clavulanic acid along with the cultivation. The highest concentration of clavulanic acid was 269.84g/L obtained under the conditions of 1.5% of soybean flour and absence of glucose in 96 hours. The strain Streptomyces parvulus DPUA 1573 was effective against multidrug-resistant strains of Staphylococcus spp. of mastitis from buffaloes, still showing concomitantly production of clavulanic acid for pharmaceutical use.

Edible mushroom species are considered as an adequate source of food in a healthy diet due to high content of protein, fiber, vitamins, and a variety of minerals. The representatives of Pleurotus genus are characterized by distinct gastronomic, nutritional, and medicinal properties among the edible mushrooms commercialized worldwide. In the present study, the growth of mycelial biomass of Pleurotus albidus cultivated in submerged fermentation was evaluated. Saccharose, fructose, and maltose were the three main carbon sources for mycelial biomass formation with corresponding yields of 7.28 g L−1, 7.07 g L−1, and 6.99 g L−1. Inorganic nitrogen sources did not stimulate growth and the optimal yield was significantly higher with yeast extract (7.98 g L−1). The factorial design used to evaluate the influence of saccharose and yeast extract concentration, agitation speed, and initial pH indicated that all variables significantly influenced the production of biomass, especially the concentration of saccharose. The greater amount of saccharose resulted in the production of significantly more biomass. The highest mycelial biomass production (9.81 g L−1) was reached in the medium formulated with 30.0 g L−1 saccharose, 2.5 g L−1 yeast extract, pH 7.0, and a speed of agitation at 180 rpm. Furthermore, P. albidus manifested different aspects of morphology and physiology under the growth conditions employed. Media composition affected mycelial biomass production indicating that the diversification of carbon sources promoted its improvement and can be used as food or supplement.

In the last years, there is a trend towards the replacement of synthetic colorants by natural ones, mainly due to the increase of consumer demand for natural products. The natural colorants are used to enhance the appearance of pharmaceutical products, food, and different materials, making them preferable or attractive. This review intends to provide and describe a comprehensive overview of the history of colorants, from prehistory to modern time, of their market and their applications, as well as of the most important aspects of the fermentation process to obtain natural colorants. Focus is given to colorants produced by filamentous fungal species, aiming to demonstrate the importance of these microorganisms and biocompounds, highlighting the production performance to get high yields and the aspects of conclusion that should be taken into consideration in future studies about natural colorants.

Made available in DSpace on 2020-12-12T01:29:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-09-15 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fungi bioproducts have recently become commercially important to the food and pharmaceutical industry. However, different structural forms exhibited by filamentous fungi throughout their life cycles are often a bottleneck for productivity in many industrial processes. In this work, the relationship among mycelium accumulation, morphology, and colorant production was evaluated. Experiments were performed in an incubator shaker and in a stirred-tank bioreactor. First, a series of nitrogen sources and pure vitamins were investigated as additives to the monosodium glutamate-glucose medium in order to improve Talaromyces amestolkiae red colorant production in the incubator shaker. MSG and pH played an important role in generating stress, which affected cell growth and colorants production. The colorant synthesis seems to be is triggered under metabolic stress. Next, the effects of starter culture composition (MSG-glucose and nitrogen complex starter media) and airflow (2.0 and 8.0 Lmin−1) were investigated in the bioreactor. After 120 h of cultivation in the bioreactor, a maximum red colorant production of 9.33 UA500nm, 72.38 % glucose consumption, and 4.53 g L-1 biomass concentration were achieved under complex nitrogen as starter medium at 8 Lmin−1. Growth remained at moderate level, probably affected by the synergistic effect of MSG and pH as stress factors. Moreover, pellets formation favored suitable oxygenation of the culture broth leading to the best consumption of the carbon source. Department of Bioprocess Engineering and Biotechnology School of Pharmaceutical Sciences Universidade Estadual Paulista - UNESP Culture Collection DPUA/UFAM Universidade Federal do Amazonas Department of Bioprocess Engineering and Biotechnology School of Pharmaceutical Sciences Universidade Estadual Paulista - UNESP

The objective of this study was to analyze the mycelial growth under different culture conditions and the antimicrobial activity of Lentinus citrinus Walleyn & Rammeloo DPUA 1535 and Neolentinus lepideus (Fr.) Redhead & Ginns DPUA 1536 against some fungi and bacteria. The growth of Lentinus spp. was evaluated in natural and complex media, with and without light, at 25°C and 28°C for 8, 15, and 30 days. To test antimicrobial activity, we used Candida albicans DPUA 1336, Cryptococcus laurentii DPUA 1501, Aspergillus flavus DPUA 1836, Escherichia coli DAUPE 224, and Mycobacterium smegmatis ATCC 607. L. citrinus showed strong growth in potato dextrose and rice meal agars in the absence and presence of light, at 28°C, and over 8, 15, and 30 days. N. lepideus grew well in Sabouraud agar in the absence and presence of light at 25°C for 8 and 15 days. L. citrinus evidenced mycelial growth and high density in potato dextrose agar, although lesser mycelial density was observed in rice bran extract agar. N. lepideus showed less abundant mycelial growth and density in the 3 culture media. Organic mycelial extracts of Lentinus species demonstrated a potential antibacterial and antifungal activity, with selectivity to the test-microorganisms under in vitro growth conditions.

The vertical mycelial growth to investigate the feasibility of Pleurotus ostreatoroseus DPUA 1720 production in lignocellulosic Amazonic residues was evaluated. Mycelial development was carried out in cupuacu exocarp (Theobroma grandiflorum Willd Former Spreng Schum), acai seed (Euterpe oleracea) and sawdust as substrates. Each residue was supplemented with rice bran, crown and pineapple peel. The average speed of mycelial growth was determined using three replicates for 15 days at 25°C in the absence and presence of light and mycelial vigor and density were evaluated. Five replicates of the selected substrate were used in mushroom production. Vigorous mycelium and strongly dense growth were observed in cupuacu exocarp treatment supplemented with rice bran. The biological efficiency, production rate and productivity were 22.90, 54.33 and 3.55%, respectively in this substrate. The basidiomata showed low levels of minerals and fat and can be considered as a source of protein (23.53%) and fiber (12.79%). Key words: basidiomata, edible mushroom, agro-industrial wastes.

Safety concerns related to the increasing and widespread application of synthetic coloring agents have increased the demand for natural colorants. Fungi have been employed in the production of novel and safer colorants. In order to obtain the colorants from fermented broth, suitable extraction systems must be developed. Aqueous two-phase polymer systems (ATPPS) offer a favorable chemical environment and provide a promising alternative for extracting and solubilizing these molecules. The aim of this study was to investigate the partitioning of red colorants from the fermented broth of Penicillium purpurogenum using an ATPPS composed of poly(ethylene glycol) (PEG) and sodium polyacrylate (NaPA). Red colorants partitioned preferentially to the top (PEG-rich phase). In systems composed of PEG 6,000 g/mol/NaPA 8,000 g/mol, optimum colorant partition coefficient (KC) was obtained in the presence of NaCl 0.1 M (KC = 10.30) while the PEG 10,000 g/mol/NaPA 8,000 g/mol system in the presence of Na2SO4 0.5 M showed the highest KC (14.78). For both polymers, the mass balance (%MB) and yield in the PEG phase (%ηTOP) were close to 100 and 79%, respectively. The protein selectivity in all conditions evaluated ranged from 2.0–3.0, which shows a suitable separation of the red colorants and proteins present in the fermented broth. The results suggest that the partitioning of the red colorants is dependent on both the PEG molecular size and salt type. Furthermore, the results obtained support the potential application of ATPPS as the first step of a purification process to recover colorants from fermented broth of microorganisms. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1295–1304, 2015

O beneficiamento do pirarucu (Arapaima gigas) gera grande quantidade de resíduos que podem ser aproveitados para a elaboração de novos produtos de interesse industrial. Este estudo objetivou avaliar a produção de hidrolisados proteicos a partir de resíduos de pirarucu, utilizando-se proteases de Aspergillus flavo-furcatis e pancreatina comercial, além de caracterizálos quanto às suas qualidades nutricional e microbiológica. A matéria-prima utilizada foi carne mecanicamente separada de carcaças de pirarucu (CMSP). Dois produtos foram desenvolvidos: um hidrolisado proteico de pirarucu produzido com enzima comercial (HPPEC) e outro com enzima microbiana (HPPEM). A CMSP e os hidrolisados foram submetidos à análise de composição química, qualidade microbiológica, grau de hidrólise, digestibilidade e perfil de aminoácidos. Os resultados mostraram que, no HPPEC, o teor proteico foi de 73,47 %, significativamente superior ao do HPPEM (58,03 %). Todavia, os dois produtos apresentaram altos valores de digestibilidade, ausência de contaminantes microbianos e redução no conteúdo de lipídios. Entre as enzimas utilizadas, a pancreatina foi a mais eficiente na elaboração do produto final, que demonstrou conteúdo de aminoácidos essenciais superior aos requerimentos para humanos adultos. O hidrolisado desenvolvido com as enzimas de A. flavo-furcatis apresentou escore de aminoácidos essenciais inferior a 1,0, sendo o aminoácido mais limitante o triptofano. The processing of arapaima (Arapaima gigas) generates a lot of residues that can be used for the development of new products of industrial interest. This study aimed at evaluating the production of protein hydrolysates from arapaima residues using Aspergillus flavo-furcatis protease and commercial pancreatin, as well as characterizing their nutritional and microbiological qualities. The raw material used was meat mechanically separated from arapaima carcasses (MMSA). Two products were developed: a protein hydrolysate of arapaima using a commercial enzyme (PHACE) and another one using microbial enzyme (PHAME). The MMSA and the hydrolysates were analyzed for chemical composition, microbiological quality, degree of hydrolysis, digestibility and amino acid profile. The results showed that the PHACE protein content was 73.47 %. This value was significantly higher, when compared to the PHAME (58.03 %). However, both products showed high digestibility values, absence of microbial contaminants and reduced lipid content. Among the enzymes used, pancreatin was the most efficient one in the preparation of the final product, which showed essential amino acids content higher than the requirements for human adults. The hydrolysate developed using A. flavo-furcatis enzymes presented essential amino acids score lower than 1.0, being tryptophan the most limiting one.

Summary Six strains of Aspergillus flavo furcatis were screened to investigate milk-clotting enzyme production by fermentation in natural liquid medium. The growth media comprised extracts of cupuacu exocarp+rice bran [10% or 20% (v/v) CE+RB] and acai waste+rice bran [10% or 20% (v/v) AW+RB] with or without supplementation of 0.1% (w/v) yeast extract and 0.5% (w/v) gelatin. Significant values of milk-clotting activity were determined by A. flavo furcatis DPUA 1461 and DPUA 1608, in the standard and natural media, respectively. According to criteria of clot and whey formations, 8.3% of the samples tested were classified as strong coagulation, 41.70% showed weak coagulation and in 50% was not observed milk coagulation. The enzyme optimal action of DPUA 1608, the selected strain, was at 40 °C and pH 7.0. Milk-clotting proteases were inhibited by pepstatin (94.72%) and moderately inhibited by the others metal ions tested.

The current study aims to identify Streptomyces spp., isolated in the Amazon region and capable of producing chitinase as well as to partially characterize the enzyme. Optimum temperature and pH, thermal and pH stabilities and the behavior of chitinase were determined as compared to other substances. Streptomyces owasiensis was the best chitinase producer in submerged culture fermentation using chitin 1% (w/v) at 140 rpm for 96 h at 34°C. The enzyme showed optimal activity at pH 7.0 and stability at the assessed pH variations. It also showed optimal temperature of 80°C and 180 min of thermostability between 30 and 90°C. The enzymatic activity was potentiated in the presence of various ions, especially Fe2+, the same occurred in the presence of the anionic surfactant sodium dodecyl sulfate (SDS), but it suffered inhibition influence by EDTA. The chitinase produced by S. owasiensis showed characteristics of industrial relevance and it highlights the first report on enzyme production from the species isolated in the Amazon. Key words: N-acetyl-D-glucosamine, chitin, chitinase, actinomycetes, Streptomyces owasiensis.

This work aims at investigating the production of yellow, orange, and red natural colorants in a submerged culture of Penicillium purpurogenum DPUA 1275. For this purpose, different experimental conditions evaluating the effect of incubation time, type and size of inoculum, and different carbon and nitrogen sources were performed. Furthermore, the growth kinetics were obtained in the conditions of 10(8) spores/ml and 5 mycelia agar discs during 360 h. These experiments showed that 5 mycelia agar discs and 336 h promoted the highest yellow (3.08 UA400nm), orange (1.44 UA470nm), and red (2.27 UA490nm) colorants production. Moreover, sucrose and yeast extract were the most suitable carbon and nitrogen sources for natural colorants production. Thus, the present study shows a new source of natural colorants, which can be used as an alternative to others available in the market after toxicological studies.

Made available in DSpace on 2018-12-11T16:41:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-05-11 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) There is a worldwide interest in the development of processes for producing colorants from natural sources. Microorganisms provide an alternative source of natural colorants produced by cultivation technology and extracted from the fermented broth. The aim of the present work was to study the recovery of red colorants from the fermented broth of Talaromyces amestolkiae using the technique of colloidal gas aphrons (CGA) comprising surfactant-stabilized microbubbles. Preliminary experiments were performed to evaluate the red colorants' solubility in different organic solvents, octanol/water partitioning, and their stability in surfactant solutions, namely hexadecyl trimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), and polyoxyethylenesorbitan monolaurate (Tween 20), which are cationic, anionic and nonionic surfactants, respectively. The first recovery experiments were carried out using CGA generated by these surfactants at different volumetric ratios (VR, 3-18). Subsequently, two different approaches to generate CGA were investigated at VR values of 6 and 12: the first involved the use of CTAB at pH 6.9-10.0, and the second involved the use of Tween 20 using red colorants partially dissolved in ethanol and Tween 20. The characterization results showed that red colorants have a hydrophilic nature. The highest recoveries were obtained with Tween 20 (78%) and CTAB (70%). These results demonstrated that the recovery of the colorants was driven by both electrostatic and hydrophobic interactions. The VR was found to be an important operating parameter and at VR 12 with CTAB (at pH 9) maximum recovery, partitioning coefficient (K = 5.39) and selectivity in relation to protein and sugar (SP = 3.75 and SS = 7.20 respectively) were achieved. Furthermore, with Tween 20, the separation was driven mainly by hydrophobic interactions. Overall CGA show promise for the recovery of red colorants from a fermented broth. Although better results were obtained with CTAB than with Tween 20 the latter may be more suitable for some application due to its lower toxicity. Department of Bioprocess and Biotechnology School of Pharmaceutical Sciences UNESP Univ. Estadual Paulista, Rodovia Araraquara-Jaú/km 01 Department of Biochemical and Pharmaceutical Technology University of São Paulo, Avenida Prof. Lineu Prestes 580, Bl. 16 Culture Collection DPUA UFAM Universidade Federal Do Amazonas, Av. Gal. Rodrigo Octávio Jordão Ramos, 3000 Department of Food and Nutritional Sciences University of Reading, Whiteknights Department of Bioprocess and Biotechnology School of Pharmaceutical Sciences UNESP Univ. Estadual Paulista, Rodovia Araraquara-Jaú/km 01 FAPESP: 0321/11-7 FAPESP: 08/58280-0

The population interest in health products is increasing day-by-day. Thus, the demand for natural products to be added in food and pharmaceutical commodity is also rising. Among these additives, colorants, which provides color to products, can be produced by microorganism through bioprocess. Looking for new source of natural colorants, fungi have been employed to this purpose producing novel and safer natural colorants. So, the main goal of this study was to describe a Talaromyces species able to produce natural colorants and investigate nutritional parameters of colorants production using statistical tool. The taxonomy classified the microorganism as Talaromyces amestolkiae. The statistical design evaluated pH and glucose, meat extract and meat peptone concentration as independent variables, and red colorants production as main response. Under the best condition (g/L: glucose 30, meat extract 1, meat peptone 10, and initial pH of 7.0) an increase of 229% in the red colorant production was achieved as compared with the initial media used. The dried fermented broth containing red colorants showed low cytotoxicity against fibroblasts cells (IC50 > 187.5 g/L) and effective antimicrobial activity against S. aureus (MIC of 2.5 g/L). Thus, T. amestolkiae colorants can be attractive to food and pharmaceutical applications as it does not produce toxic compounds and can promote protection against microorganism contaminants. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2684, 2019.

Objective: The ASINPHAR@2action programme aims at raising awareness to early vascular ageing (EVA) through a community-based intervention. This preliminary analysis is focused on the analysis of the proportion of participants with abnormal arterial stiffness (AS) and the definition of its main determinants. Design and method: This preliminary analysis is a cross-sectional, observational, descriptive, non-interventional study of participants enrolled in 11 communal pharmacies in Portugal (HOLON pharmacies), between April and November 2017. Blood pressure (BP) and arterial function parameters were measured with a non-invasive validated device (MOBIL-O-GRAPH, IEM®). Clinical and demographic information was gathered. Results: Participants recruited for the project account for 658, 65.7% women, with a mean age of 57.34 ± 16.26 years (range: 20–96 years). Brachial BP was 126.60 ± 16.43 mmHg and 79.89 ± 11.54 mmHg, and central BP was 115.80 ± 15.35 mmHg and 81.18 ± 11.60 mmHg, respectively for systolic and diastolic BP. Mean pulse wave velocity (PWV) was 8.45 ± 2.28 m/s. The proportion of participants with increased PWV was 19,9%. Participants with increased PWV were significantly older and had higher brachial and central BP and BMI. Multivariate linear regression indicated age, Gender, BP and abdominal fat as independent determinants of PWV. AS trajectories were significantly different as a function of arterial hypertension and cardiovascular risk classification (figure 1). Conclusions: The preliminary results of this pioneering large scale study measuring arterial function in communal pharmacies provides the grounds for the operationalization of subclinical target organ damage screening in pharmacies, as a strategy to improve cardiovascular risk monitoring and to promote adherence to treatment.

Brosimum parinarioides, membro da família Moraceae, espécie vegetal nativa da região norte conhecida como “amapazeiro” é fonte de látex, produto tradicionalmente consumido como alimento e usado na medicina popular por comunidades para o tratamento de patologia pulmonar e como tonificante. Embora o látex desse vegetal seja consumido como alimento funcional e tenha importância industrial farmacêutica, existem poucos estudos a cerca da avaliação das propriedades dos compostos bioativos presentes neste líquido. O objetivo deste trabalho foi avaliar a composição nutricional e analisar a qualidade do produto por dois métodos de extração do látex visando orientar os agricultores extrativistas para aquisição de um alimento com qualidade para consumo humano. As amostras de látex foram submetidas à análise da composição centesimal, toxicidade e determinação da qualidade microbiológica. Os resultados demonstraram que o método de assepsia mais eficiente para a qualidade microbiológica do produto foi o uso da aspersão de hipoclorito de sódio seguido de álcool no caule da árvore. O látex apresentou compostos bioativos com propriedades antioxidante, não apresentou efeito hemolítico em sangue de carneiro e nem toxicidade frente a Artemia salina. Os resultados de composição centesimal demonstram que este é um produto altamente energético, justificando seu uso como alimento matinal pelas populações da Amazônia.