Specific proteases capable of degrading native triple helical or denatured collagen have been required for many years and have a large spectrum of applications. There are few complete reports that fully uncover production, characterization and purification of fungi collagenases. In this review, authors searched through four scientific on line data bases using the following keywords (collagenolytic OR collagenase) AND (fungi OR fungus OR fungal) AND (production OR synthesis OR synthesize) AND (characterization). Scientific criteria were adopted in this review to classify found articles by score (from 0 to 10). After exclusion criteria, 21 articles were selected. None obtained the maximum of 10 points defined by the methodology, which indicates a deficiency in studies dealing simultaneously with production, characterization and purification of collagenase by fungi. Among microorganisms studied the non-pathogenic fungi Penicillium aurantiogriseum and Rhizoctonia solani stood out in volumetric and specific collagenase activity. The only article found that made sequencing of a true collagenase showed 100% homology with several metalloproteinases fungi. A clear gap in literature about collagenase production by fungi was verified, which prevents further development in the area and increases the need for further studies, particularly full characterization of fungal collagenases with high specificity to collagen.
Collagenases are proteolytic enzymes capable of degrading both native and denatured collagen, reported to be applied in industrial, medical and biotechnological sectors. Liquid-liquid extraction using aqueous two-phase system (ATPS) is one of the most promising bioseparation techniques, which can substitute difficult solid-liquid separation processes, offering many advantages over conventional methods including low-processing time, low-cost material and low-energy consumption. The collagenase produced by Penicillium sp. UCP 1286 showed a stronger affinity for the bottom salt-rich phase, where the highest levels of collagenolytic activity were observed at the center point runs, using 15.0% (w/w) PEG 3350 g/mol and 12.5% (w/w) phosphate salt at pH 7.0 and concentration. The enzyme was characterized by thermal stability, pH tolerance and effect of inhibitors, showing optimal collagenolytic activity at 37 °C and pH 9.0 and proved to be a serine protease. ATPS showed high efficiency in the collagenase purification, confirmed by a single band in SDS/PAGE, and can in fact be applied as a quick and inexpensive alternative method.
This work reports an optimization of protease from Penicillium aurantiogriseum immobilization on polyaniline-coated magnetic nanoparticles for antioxidant peptides' obtainment derived from bovine casein. Immobilization process was optimized using a full two-level factorial design (2) followed by a response surface methodology. Using the derivative, casein was hydrolyzed uncovering its peptides that were sequenced and had antioxidant properties tested through (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) (ABTS) radical scavenging and hydrogen peroxide scavenging assays. Optimal conditions for immobilization were 2 hr of immobilization, offered protein amount of 200 µg/mL, immobilization pH of 6.3 and 7.3 hr of activation. Derivative keeps over 74% of its original activity after reused five times. Free and immobilized enzyme casein hydrolysates presented similar peptide mass fingerprints, and prevalent peptides could be sequenced. Hydrolysates presented more than 2.5× higher ROS scavenging activity than nonhydrolyzed casein, which validates the immobilized protease capacity to develop casein-derived natural ingredients with potential for functional foods.
O envelhecimento feminino representa um desafio de priorizar medidas de prevenção e promoção da saúde, e se destaca a atenção ao climatério e menopausa. O presente trabalho se propôs a conhecer e analisar o nível de informação das mulheres na fase do climatério, atendidas numa unidade de saúde da família (USF) de Recife - PE. Os dados foram obtidos através de entrevista estruturada aplicada após a consulta seguindo todas diretrizes de ética em pesquisa. As respostas das 20 voluntárias, de 45 a 79 anos, evidenciaram o perfil destas mulheres, os sintomas e problemas mais frequentes e quais as principais mudanças identificadas nesta fase. A maioria das mulheres relatou vivenciar ondas de calor e sintomas psicológicos, como mal-estar e irritação. Além de relacionarem a fase do climatério com sentimentos negativos. Foram evidentes a desinformação das usuárias e a necessidade de melhor suporte e orientação por parte dos profissionais de saúde. Ainda, ficou claro a importância de valorizar a influência da família sobre a adaptação, a ser considerada pela equipe multidisciplinar. Diversificadas foram as experiências relatadas, reafirmando que cada mulher vive esta fase de maneira individual, assim como deve ser a adequação de seu acolhimento, de forma individualizada, integrada e respeitosa.
Uso de substratos agroindustriais para produção de delta-endotoxinas por Bacillus thuringiensis Use of agro-industrial substrates for delta-endotoxins production by Bacillus thuringiensis
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