Integrated process production and extraction of the fibrinolytic protease from Bacillus sp. UFPEDA 485

Fibrinolytic proteases are enzymes that degrade fibrin; these enzymes are a promising alternative for thrombolytic therapy, and microorganisms produce them. The aim of this study was to evaluate the optimum conditions for the integrated production and purification of fibrinolytic protease from Bacillus sp. UFPEDA 485. Extractive fermentation was carried out in a culture medium containing soybean flour and by adding polyethylene glycol (PEG) and Na2SO4 according to a 23 experimental design. In all assays, the enzyme preferentially partitioned to the bottom phase (K < 1), with an optimum activity of 835 U ml−1 in the bottom phase (salt-rich phase). The best conditions for extractive fermentation were obtained with 18 % PEG 8000 and 13 % Na2SO4. Characterization showed that it is a metalloprotease, as a strong inhibition—residual activity of 3.13 %—occurred in the presence of ethylenediaminetetraacetic acid. It was also observed that enzymatic activity was stimulated in the presence of ions: CaCl2 (440 %), MgCl2 (440 %), FeSO4 (268 %), and KCl (268 %). The obtained results indicate that the use of a low-cost substrate and the integration of fermentation with an aqueous two-phase system extraction may be an interesting alternative for the production of fibrinolytic protease.The authors thank CAPES (National Council for the Improvement of Higher Education) for the scholarship and CNPq (National Counsel of Technological and Scientific Development) and RENORBIO for the financial support

Evaluation the best condition of Fibrinolytic protease production using factorial design by Streptomyces sp DPUA 1573

XI Reunião Regional Nordeste da SBBq | 4th International Symposium in Biochemistry of Macromolecules and BiotechnologyFibrinolytic enzymes have the ability to degrade fibrin clots formed for avoiding intravascular thrombosis. In the pharmaceutical industry there is a search for new fibrinolytic agent that reduces the production cost and increasing productivity. The use of microorganism for enzyme production, such as the genus Streptomyces has been reported. Streptomyces is a Gram-positive bacteria, responsible for producing many bioactive compounds and extracellular enzymes of pharmaceutical interest. This study aimed to evaluated the production of fibrinolytic protease by Streptomyces sp DPUA 1573. Microbial cells were cultivated in the ISP2 for 48 hours, after this period the strains were inoculated in MS2 (soybean medium) that according with factorial design 24 (concentrations of soybean 0.5; 1.0 and 1.5%, glucose 0; 0.5 and 1.0% and different speeds 150 rpm; 200 rpm and 250 rpm and temperature 28C; 30C and 32C). The factorial design was analyzed by variance analysis (anova) and the glucose concentration showed a positive and significative effect. The results showed that the variable interaction had significant effect. that the best condition was composed 1.5% soybean, 1% glucose, 28 ºC and 150 speed in 48 hours, with production fibrinolytic 1391.66 U/mL. These values were higher than those reported in the literature. However these results show the biggest potencial in production fibrinolytic enzyme by Streptomyces.info:eu-repo/semantics/publishedVersio

Production and characterization of new fibrinolytic protease from Mucor subtillissimus UCP 1262 in solid-state fermentation

Fibrinolytic enzymes have received attention regarding their medicinal potential for thrombolytic diseases, a leading cause of morbidity and mortality worldwide. Various natural enzymes purified from animal, plant and microbial sources have been extensively studied. The aim of this work was to produce fibrinolytic protease by solid state fermentation using agro industrial substrates. Rhizopus arrhizus var. arrhizus UCP 1295 and Mucor subtillissimus UCP 1262 filamentous fungi species isolated from soil of Caatinga-PE, Brasil, were used as producer microorganisms. Wheat bran was shown to be the best substrate for the production of the enzyme and by using a 23 full factorial design the main effects and interactions of the quantity of the substrate wheat bran, moisture and temperature on the fibrinolytic enzyme production and protease were evaluated. The best results for fibrinolytic and protease activities, 144.58 U/mL and 48.33 U/mL, respectively, were obtained with Mucor subtillissimus UCP 1262 using as culture medium 3 g wheat bran, 50% moisture at a temperature of 25˚C for 72 hours. The optimum temperature for the produced enzyme was 45˚C and most of its original activity was retained after being subjected to 80˚C for 120 min. The protease activity was enhanced by K+, Ca+ and Mn+; but with Cu+ there was an inhibition. The specificity to chromogenic substrate and the inhibition by PMSF indicates that it is a chymotrypsin-like serine protease. Presented results suggest that this enzyme produced by solid-state fermentation is an interesting alternative as a candidate for thrombolytic therapy

Effect of pH and temperature on phytase and biomass production by submerged fermentation with Aspergillus niger var. phoenicis URM 4924

Phytase production and biomass was evaluated in present work by submerged fermentation with Aspergillus niger var. phoenicis URM 4924. Experimental assays were done under different conditions of pH (4.0 to 8.0) and temperature (25 to 35 ºC), and the influence of these variables on the responses was studied through a 22 central composite design and response surface methodology. Phytase and biomass production were affected by the pH and temperature used during submerged fermentation. Phytase activity was increased in up to 7.8-fold (from 1.04 to 8.09 U/mL) and the ergosterol content was increased in up to 38-fold (from 9.3 to 354.09 μg/mL). The maximum values of both responses were achieved when using pH 4.0 and 30 ºC

Recovery and partitioning of fibrinolytic protease from Bacillus sp. UFPEDA 485 by aqueous two-phase systems

Fibrinolytic proteases produced by Bacillus sp. has attracted interest in the pharmaceutical industry as a promising alternative in thrombolytic therapy due to their effectiveness in degrading fibrin, its production requiring the development of an efficient recovery process. Aqueous two-phase systems (ATPS) have been recognized as an efficient and economical process for recovering enzymes. To optimize the recovery of fibrinolytic protease from the fermentation broth of Bacillus sp. UFPEDA 485, a 23 full factorial design was used to evaluate the influence of the three independent variables PEG molar mass (MPEG), PEG concentration (CPEG) and sodium sulfate concentration (CNa2SO4) on the partition coefficient (K), purification factor (PF) and yield recovery (Y) of fibrinolytic protease in PEG/Na2SO4 aqueous two-phase system. For all ATPS studied, enzymes partitioned to the top phase and the highest extraction was obtained for MPEG 6000 g.mol-1, CPEG 24 % (w/w) and CNa2SO4 11.6 % (w/w) with K = 5.03; PF = 3.30; Y = 91.40% and Fibrinolytic activity in the top phase 821 U.mL-1. Findings reported here show that ATPS composed of PEG/Na2SO4 is a valuable strategy for the extraction of fibrinolytic protease and can be considered a promising method for the extraction of enzymes in industrial scale

Regulação em saúde e epistemologia política: contribuições da Ciência Pós-Normal para enfrentar as incertezas

Este estudo te&#243;rico-conceitual considera a regula&#231;&#227;o em sa&#250;de como uma a&#231;&#227;o complexa que lida com uma pluralidade de perspectivas em situa&#231;&#245;es com in&#250;meras incertezas existentes. A regula&#231;&#227;o em sa&#250;de possui finalidade de proteger a sa&#250;de da popula&#231;&#227;o na defesa do interesse p&#250;blico, sendo a dimens&#227;o da qualidade do conhecimento central para a tomada de decis&#245;es. Nessa perspectiva, o artigo possui como objetivo discutir as contribui&#231;&#245;es te&#243;rico-metodol&#243;gicas da Ci&#234;ncia P&#243;s-Normal para tal regula&#231;&#227;o, abordagem que aprofunda e prop&#245;e estrat&#233;gias de gest&#227;o de temas complexos relacionados &#224; interface ci&#234;ncia-pol&#237;tica. Destaca aspectos que a ci&#234;ncia tradicional tende a desconsiderar como as incertezas, o peso dos valores e a pluralidade de perspectivas leg&#237;timas frente &#224; qualidade dos tomadas de decis&#227;o, justificando a relev&#226;ncia dos processos participativos em uma perspectiva democr&#225;tica e epistemol&#243;gica. Conclui-se que a incorpora&#231;&#227;o das estrat&#233;gias metodol&#243;gicas da Ci&#234;ncia P&#243;s-Normal pode contribuir para a atividade regulat&#243;ria em sa&#250;de por integrar epistemologia e pol&#237;tica, ampliando a qualidade das decis&#245;es regulat&#243;rias

Monitoramento tecnológico da aplicação de enzimas celulolíticas: panorama mundial e brasileiro

As celulases são enzimas capazes de atuar sobre materiais celulósicos, promovendo sua hidrólise. Estas enzimas apresentam importância na pesquisa a nível acadêmico e industrial devido às suas aplicações que contribuem em vários processos industriais, como nas indústrias de papel e celulose, na produção de bioetanol, na indústria de alimentos, gestão de resíduos, entre outros. Desta forma, o presente trabalho teve como objetivo a realização de um monitoramento tecnológico de patentes sobre aplicações das celulases. Após a coleta de informações nas bases de dados (Lens, Derwent Inovation Index, Espacenet, Patentscope, USPTO e INPI), observou-se que o Derwent apresentou um resultado mais amplo de registro de patentes. A maioria das patentes foi agrupada na área de conhecimento química e entre os depositantes, a empresa Novozymes apresentou a maior quantidade de documentos. Ao avaliar o cenário nacional, verificou-se que a maioria das patentes depositadas no INPI são de depositantes estrangeiros e em termos institucionais, a grande maioria das patentes é proveniente de instituições privadas. Havendo uma necessidade de maior investimento em pesquisa nas universidades e institutos de pesquisa governamentais objetivando fomentar a inovação em aplicações industriais de enzimas, especialmente celulases, que é uma área de grande importância econômica

Medidas de contenção de tipo lockdown para prevenção e controle da COVID-19: estudo ecológico descritivo, com dados da África do Sul, Alemanha, Brasil, Espanha, Estados Unidos, Itália e Nova Zelândia, fevereiro – agosto de 2020

Objective: To describe the lockdown measures and incidence of COVID-19 in South Africa, Germany, Brazil, Spain, United States, Italy and New Zealand. Methods: The descriptive ecological study with data on the daily incidence of confirmed cases of COVID-19, from February 22 to August 31, 2020 on COVID-19, and information on lockdown containment measures implemented by the governments of each country. Results: The countries that implemented lockdown, from the beginning of its implementation until three weeks later, had a decrease in the incidence (in cases per million inhabitants): South Africa (3.7 to 1.7), Germany (37.5 to 33.7) Spain (176.3 to 82.0), Italy (92.0 to 52.1) and New Zealand (7.5 to 1.7). As for Brazil and the United States, which did not implement lockdown, there was no considerable decrease. Conclusion: After the implementation of lockdown, there was a considerable decrease in the number of confirmed cases.Objetivo: Descrever as medidas de contenção de tipo lockdown e a incidência da COVID-19 em sete países: África do Sul, Alemanha, Brasil, Espanha, Estados Unidos, Itália e Nova Zelândia. Métodos: Estudo ecológico descritivo, com dados da incidência diária dos casos confirmados de COVID-19 entre 22 de fevereiro e 31 de agosto de 2020, e informações sobre medidas de lockdown implementadas pelos governos de cada país. Resultados: Os países que implementaram lockdown tiveram diminuição da incidência diária de COVID-19 (casos por milhão de habitantes) no período de três semanas, a contar do início da medida: África do Sul (3,7 a 1,7), Alemanha (37,5 a 33,7), Espanha (176,3 a 82,0), Itália (92,0 a 52,1) e Nova Zelândia (7,5 a 1,7). O Brasil e os Estados Unidos, que não implementaram lockdown, não apresentaram uma diminuição considerável. Conclusão: Após a implementação de lockdown, houve uma diminuição considerável do número de casos confirmados

Avaliação de variáveis que influenciam a hidrólise enzimática da caseína do leite de cabra Moxotó

The aim of this work was to evaluate the effects of temperature, pH, enzyme:substrate ratio (E:S), and reaction time on the enzymatic hydrolysis of Moxotó breed goat's milk casein, using different proteolytic enzymes. Enzymatic hydrolysis of the capric casein was performed using trypsin, pepsin and papain. For each enzyme, two levels of the variables were used to assess their effects on the hydrolysis of casein, using a full factorial design (24). The hydrolysis products were visualized on SDS-PAGE. The best hydrolysis degree using papain were at pH 6.5, E:S of 1:150 and 5 hours of hydrolysis at 50°C (28.17%); for trypsin, at pH 8.5, E:S of 1:150 and 5 hours at 40°C (29.55%); and for pepsin, at pH 3.0, E:S of 1:100 and 5 hours at 40°C (38.27%). Capric casein hydrolysis is affected positively by pH and reaction time, using pepsin. Significant antagonistic interactions were observed between pH and E:S, with trypsine; and between pH, temperature and reaction time, with papain. Pepsin has high αs1, β e κ-casein hydrolysis efficiency, evidenced by the molar masses below 14.4 kDa of its products.O objetivo deste trabalho foi avaliar os efeitos da temperatura, pH, relação enzima:substrato (E:S) e tempo de reação sobre a hidrólise enzimática da caseína do leite de cabra Moxotó, com uso de diferentes enzimas proteolíticas. A hidrólise enzimática da caseína caprina foi realizada com uso de tripsina, pepsina e papaína. Para cada enzima, foram utilizados dois níveis de cada variável, na avaliação de seus efeitos sobre a hidrólise da caseína, em um fatorial completo 24. Os produtos da hidrólise foram visualizados em eletroforese SDS-PAGE. O melhor valor do grau de hidrólise com a enzima papaína foi obtido em pH 6,5, E:S de 1:150 e 5 horas de hidrólise a 50ºC (28,17%); com a tripsina, em pH 8,5, E:S de 1:150 e 5 horas a 40ºC (29,55%); e com a pepsina, em pH 3,0, E:S de 1:100 e 5 horas a 40ºC (38,27%). A hidrólise da caseína caprina é influenciada positivamente pelo pH e tempo de hidrólise, com o uso da pepsina. Foram observadas interações significativas antagônicas entre pH e relação E:S, com o uso da tripsina; e entre pH, temperatura e tempo de hidrólise, com o uso da papaína. A pepsina apresenta elevada eficiência na hidrólise de αs1, β e κ-caseína, evidenciada por hidrolisados com massa molar abaixo de 14,4 kDa