Purification of an Exopolygalacturonase from Penicillium viridicatum RFC3 Produced in Submerged Fermentation

An exo-PG obtained from Penicillium viridicatum in submerged fermentation was purified to homogeneity. The apparent molecular weight of the enzyme was 92 kDa, optimum pH and temperature for activity were pH 5 and 50–55°C. The exo-PG showed a profile of an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of pectin with a high degree of esterification (D.E.). Ions Ca2+ enhanced the stability of enzyme and its activity by 30%. The Km was 1.30 in absence of Ca2+ and 1.16 mg mL−1 in presence of this ion. In relation to the Vmax the presence of this ion increased from 1.76 to 2.07 μmol min−1mg−1

Cultivation of edible mushroom Hiboukitake in caja bagasse by in Jun-Cao technique

A adaptação das espécies de Pleurotus a novos resíduos representa atualmente um dos principais processos de bioconversão de resíduos agroindustriais em produtos comestíveis de alto valor nutricional. Com o presente trabalho, utilizando-se do princípio das tecnologias limpas, cultivou-se a linhagem/lote EF-133/11 de Pleurotus sajor-caju em bagaço de cajá suplementado com bagaço de cana-de-açúcar. A eficiência biológica (EB) obtida no Tratamento 1 (T1 -composto de 70% bagaço de cana e 30% do bagaço de cajá) foi de 28,94 ± 3,62 e no Tratamento 2 (T2 -composto por 50% do bagaço de cana e 50% de bagaço de cajá) foi 26,37±5,01. Tais valores não apresentaram diferença estatística para ointervalo de confiança de 95%, sendo as relações C/N de 47,14:1 e 52,00:1, respectivamente. Contudo, percebe-se uma redução na EB de T2, mesmo tendo uma alta relação C/N em comparação com T1, característica esta que provavelmente é devido ao fato do cajá ser um fruto carnoso, além da presença de bastante pectina, o que confere ao bagaço uma característica mucilaginosa dificultando as trocas gasosas no substrato e causando compactação. A padronização do cultivo em bagaço de cajá permitirá uma EB razoável podendo ser utilizado como substrato para futuros cultivos comercial.Adapting to new waste to cultivate species of Pleurotus is currently one of the main processes of bioconversion of agro-industrial residues in edible products of high nutritional value. The present work, using the principle of clean technologies, cultured strain / lot EF-133/11 of Pleurotus sajor-caju on bagasse caja supplemented with crushed sugar cane. The biological efficiency (BE) obtained in Treatment 1 (T1 -composed of 70% bagasse sugarcane and 30% of bagasse caja) was 28.94 ± 3.62 and in Treatment 2 (T2 –composed of 50% bagasse sugarcane and 50% of bagasse caja) was 26.37 ± 5.01, these were not statistically different for the confidence interval of 95%, and the relationships C / N 52.00:1and 47.14:1, respectively. However it is perceived in a decay of BE T2, even with a high C / N ratio compared to T1, this feature is probably due to the fact that being a caja fruit, rather than the presence of pectin, which gives the bagasse a characteristic mucilaginous hindering gas exchange in the substrate and causing compaction. The standardization of cultivation bagasse cajá allows a reasonable BE can be used as a substrate for future commercial crops

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Purificação, carcterização físico-química e termodinâmica de ß-glicosidases produzidas pelos microrganismos Aureobasidium pullulans e Thermoascus aurantiacus: aplicação em isoflavonas e terpenos glicosilados

Neste trabalho as ß-glicosidases produzidas pelos microrganismos Thermoascus aurantiacus e Aureobasidium pullulans foram purificadas, caracterizadas termodinamicamente, físico-quimicamente e aplicadas em terpenos e isoflavonas. O processo de purificação da enzima do T. aurantiacus apresentou rendimento final de 63,6% e fator de purificação de 46,3 vezes. Após a purificação da enzima do A. pullulans, esta apresentou fator de purificação de 35,5 vezes e rendimento de 19%. A ß-glicosidase do T. aurantiacus apresentou atividade ótima em pH 4,5 a temperatura de 75ºC. Esta manteve-se estável na faixa de pH 4,5 a 6,5 e reteve sua atividade original após 1 hora a 70ºC. A enzima purificada do A. pullulans apresentou maior atividade catalítica nos valores de pH 4,0-4,5 a temperatura de 80ºC. A mesma foi estável em ampla faixa de pH 4,0 a 9,5, e, reteve sua atividade original após 1 hora a 75ºC. A maior termoestabilidade, da enzima produzida pelo microrganismo mesofílico, A. pullulans, foi comprovada pela análise dos parâmetros termodinâmicos. A ß-glicosidase do T. aurantiacus apresentou valores menores de t(1/2), ∆G, ∆H e Ea em todas as temperaturas analisadas. As duas enzimas foram fortemente inibidas por Hg+2 e Ag+, sugerindo que o grupo tiol (SH) é essencial para atividade das mesmas. O pNPßG foi utilizado para a determinação dos parâmetros cinéticos, os valores de Km e Vmax obtidos foram 0,53 mM e 590,8 μmol/ min/mg de proteína para a enzima do T. aurantiacus, e, 0,93 mM e 29,9 μmol/ min/mg de proteína para a enzima do A. pullulans. A presença de etanol no meio de reação ativou a ß-glicosidase do T. aurantiacus, indicando atividade glicosil transferase, o mesmo não foi observado para enzima do A. pullulans. As duas ß-glicosidases atuaram em diferentes substratos glicosídicos, inclusive sobre terpenos e isoflavonas, o que confirma a ampla especificidade das enzimas.In this work, ß-glucosidases produced by the microorganisms Thermoascus aurantiacus and Aureobasidium pullulans were purified, thermodynamically and physical-chemically characterized and applied on terpenes and isoflavones. Purification process of the enzyme from T. aurantiacus exhibited final yield of 63.6% and purification factor of 46.3 -fold. Purification process of the enzyme from A. pullulans resulted in 19% yield and purification factor of 35.5-fold. ßglucosidase from T. aurantiacus exhibited optimum activity at pH 4.5 and at temperature of 75ºC. It remained stable at pH range of 4.5 to 6.5 and maintained its original activity after 1 hour at 70ºC. Purified enzyme from A. pullulans exhibited a higher catalytic activity in pH values of 4.0-4.5 at an optimum temperature of 80ºC. It was stable over a wide pH range, from 4.0 to 9.5, and, maintained its original activity after 1 hour at 75ºC. The higher thermal stability of the enzyme produced by the mesophilic microorganism, A. pullulans, was confirmed by analysis of the thermodynamic parameters. ß-glucosidase from T. aurantiacus exhibited lower values for t(1/2), ∆G, ∆H and Ea in all temperatures tested. Both enzymes were strongly inhibited by Hg+2 and Ag+, suggesting that a thiol group (SH) is essential for their activities. pNPßG was used for the kinetic parameters determination and the values for Km and Vmax were 0.53 mM and 590.8 μmol/ min/mg of protein for the enzyme from T. aurantiacus, and, 0.93 mM and 29.9 μmol/ min/mg of protein for the enzyme from A. pullulans. The presence of ethanol in the reaction mixture activated the ß-glucosidase from T. aurantiacus, indicating glucosil transferase activity, which was not observed for the enzyme from A. pullulans. Both ß-glucosidases exhibited activity on different glucosidic substrates, including terpenes and isoflavones, confirming the broad specificity of the enzymes.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Purification and properties of polygalacturonase produced by thermophilic fungus Thermoascus aurantiacus CBMAI-756 on solid-state fermentation

Polygalacturonases are enzymes involved in the degradation of pectic substances, being extensively used in food industries, textile processing, degumming of plant rough fibres, and treatment of pectic wastewaters. Polygalacturonase (PG) production by thermophilic fungus Thermoascus aurantiacus on solid-state fermentation was carried out in culture media containing sugar cane bagasse and orange bagasse in proportions of 30% and 70% (w/w) at 45°C for 4 days. PG obtained was purified by gel filtration and ion-exchange chromatography. The highest activity was found between pH 4.5 and 5.5, and the enzyme preserved more than 80% of its activity at pH values between 5.0 and 6.5. At pH values between 3.0 and 4.5, PG retained about 73% of the original activity, whereas at pH 10.0 it remained around 44%. The optimum temperature was 60–65°C. The enzyme was completely stable when incubated for 1 hour at 50°C. At 55°C and 60°C, the activity decreased 55% and 90%, respectively. The apparent molecular weight was 29.3 kDa, Km of 1.58 mg/mL and Vmax of 1553.1μmol/min/mg. The presence of Zn+2, Mn+2, and Hg+2 inhibited 59%, 77%, and 100% of enzyme activity, respectively. The hydrolysis product suggests that polygalacturonase was shown to be an endo/exoenzyme

Production of β-glucosidase on solid-state fermentation by Lichtheimia ramosa in agroindustrial residues: Characterization and catalytic properties of the enzymatic extract

Background: β-Glucosidases catalyze the hydrolysis of cellobiose and cellodextrins, releasing glucose as the main product. This enzyme is used in the food, pharmaceutical, and biofuel industries. The aim of this work is to improve the β-glucosidase production by the fungus Lichtheimia ramosa by solid-state fermentation (SSF) using various agroindustrial residues and to evaluate the catalytic properties of this enzyme. Results: A high production of β-glucosidase, about 274 U/g of dry substrate (or 27.4 U/mL), was obtained by cultivating the fungus on wheat bran with 65% of initial substrate moisture, at 96 h of incubation at 35°C. The enzymatic extract also exhibited carboxymethylcellulase (CMCase), xylanase, and β-xylosidase activities. The optimal activity of β-glucosidase was observed at pH 5.5 and 65°C and was stable over a pH range of 3.5–10.5. The enzyme maintained its activity (about 98% residual activity) after 1 h at 55°C. The enzyme was subject to reversible competitive inhibition with glucose and showed high catalytic activity in solutions containing up to 10% of ethanol. Conclusions: β-Glucosidase characteristics associated with its ability to hydrolyze cellobiose, underscore the utility of this enzyme in diverse industrial processes

Production and Characterization of β-glucosidase Obtained by the Solid-State Cultivation of the Thermophilic Fungus Thermomucor indicae-seudaticae N31

In this paper, several agro-industrial wastes (soybean meal and wheat straw, rice and peanut husks, corn cob and corn stover, and sugarcane bagasse) were tested for the production of β-glucosidase by the cultivation of thermophilic fungus Thermomucor indicae-seudaticae N31 in solid-state fermentation (SSF). Among the tested substrates, the highest yields were obtained in soybean meal. Other fermentation parameters were also evaluated, such as initial pH, merge substrates, and fermentation time, as well as the physicochemical characterization of the enzyme. The best results were obtained after 192 h of fermentation with the initial pH adjusted to 6.0. The substrate mixture did not improve the enzyme production by microorganism. The β-glucosidase showed best catalytic activity at pH 4.5 and at 75 °C and remained stable in the pH range from 4.5 to 9.5 and the temperature range 40–75 °C. The enzyme showed 80 % of its activity at a concentration of 15 mM glucose and remained stable up to 20 % ethanol.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Catalytic Properties of Amylolytic Enzymes Produced by Gongronella butleri Using Agroindustrial Residues on Solid-State Fermentation

Amylases catalyze the hydrolysis of starch, a vegetable polysaccharide abundant in nature. These enzymes can be utilized in the production of syrups, alcohol, detergent, pharmaceutical products, and animal feed formulations. The aim of this study was to optimize the production of amylases by the filamentous fungus Gongronella butleri by solid-state fermentation and to evaluate the catalytic properties of the obtained enzymatic extract. The highest amylase production, 63.25 U g−1 (or 6.32 U mL−1), was obtained by culturing the fungus in wheat bran with 55% of initial moisture, cultivated for 96 h at 25°C. The enzyme presented optimum activity at pH 5.0 and 55°C. The amylase produced was stable in a wide pH range (3.5–9.5) and maintained its catalytic activity for 1 h at 40°C. Furthermore, the enzymatic extract hydrolyzed starches from different vegetable sources, presenting predominant dextrinizing activity for all substrates evaluated. However, the presence of glucose was observed in a higher concentration during hydrolysis of corn starch, indicating the synergistic action of endo- and exoamylases, which enables the application of this enzymatic extract to produce syrups from different starch sources