Alex Fernando de Almeida

This study evaluated the immobilization of an invertase from Bacillus tequilensis, isolated from the fruit of the peach palm, on silica gel (SG) and polyhydroxybutyrate (PHB), aiming at its application in sucrose hydrolysis for invert sugar production. Immobilization was performed at 30 °C and pH 5.5 using glutaraldehyde-functionalized supports. Immobilization yield (IY), recovered activity (RA), kinetic parameters, and pH, thermal, storage, and operational stabilities were evaluated. IY values reached 72% (SG) and 77% (PHB), with RA of 35% and 67%, respectively. Both immobilized derivatives exhibited improved storage stability compared to the soluble enzyme. The SG-immobilized invertase demonstrated enhanced thermostability and higher fructose production and retained over 50% of its activity after three reuse cycles. In contrast, the PHB derivative showed greater substrate affinity. These results demonstrate that the B. tequilensis invertase immobilized on SG is a promising and stable biocatalyst derived from the Amazonian biodiversity, suitable for sustainable invert sugar production.

Microbial lipases are pivotal biocatalysts in the food, pharmaceutical, and bioenergy industries due to their versatility and stability. Among potential producers, the yeast Candida viswanathii has shown promise, yet the transition from laboratory shake flasks to stirred tank bioreactors often presents challenges due to altered physiological and metabolic responses. Specifically, determining how different carbon sources influence the trade-off between biomass growth and enzyme secretion during this scale-up remains a critical bottleneck. Here we show that while sorbitol acts synergistically with soybean oil to maximize lipase production in shake flasks (13.56 U mL^–1), it triggers a strong inhibitory effect on enzyme expression when scaled up to a stirred tank bioreactor. Contrary to the expectation that optimized flask conditions would translate linearly, the bioreactor environment shifted the yeast’s metabolism toward rapid biomass accumulation (30 g L^–1) at the expense of lipolytic activity, with the highest reactor yields achieved only in the absence of sorbitol (13.56 U mL^–1). These findings underscore the necessity of re-evaluating medium composition specifically for dynamic reactor environments, rather than relying solely on static screening data to predict industrial performance. It is concluded that cultivation in bioreactors for enzyme production must carefully consider agitation and aeration conditions, as these conditions modify the yeast’s behavior, altering nutrient utilization and, consequently, the induction of enzymatic expression and cell growth.

This study investigates the diversity of filamentous fungi in the Amazon biome, particularly the genus Fusarium, known for producing biotechnologically valuable metabolites. The research aimed to isolate and identify fungi from Oenocarpus bacaba Mart. fruits and analyze the biochemical properties of lipase produced by the strains (maximum activity of 1750 U mL⁻¹). Two isolates, FF1 and FF2, were identified using morphological and molecular techniques, with ITS sequence data suggesting they belong to the Fusarium fujikuroi species complex, though the exact species remains unconfirmed. This is the first report that highlights the biotechnological potential of the F. fujikuroi complex isolated from Oenocarpus bacaba, emphasizing the relevance of Amazonian biodiversity as a source of microorganisms with promising applications in sustainable industrial processes. The results show that native fruits, such as bacaba, are effective matrices for prospecting filamentous fungi producers of enzymes of biotechnological interest, such as lipases. These findings reinforce the importance of rational exploitation of the Amazonian microbiota for the development of bioproducts and eco‐efficient industrial processes.

Publicado em 04 de outubro de 2023. As ciências biológicas têm sido a vanguarda de uma jornada de descobertas que remodelaram profundamente nossa visão da vida e do mundo natural. Do micromundo das estruturas moleculares ao macrocosmo dos ecossistemas complexos, essa abrangente disciplina tem construído uma base sólida para avanços que ecoam positivamente em toda a sociedade. Ela se estabelece como a força motriz que impulsiona o progresso, deixando uma marca indelével em nosso planeta. Esses avanços reverberam em melhorias substanciais na saúde humana, na nutrição, na qualidade do ambiente e na nossa compreensão intrínseca da vida. À medida que exploramos continuamente as fronteiras da biologia, traçamos um caminho luminoso em direção a um futuro mais saudável e sustentável, onde a ciência se torna a luz que guia nosso desenvolvimento. Com grande prazer, apresentamos o segundo volume do E-Book "Avanços nas Ciências Biológicas e seus Impactos na Sociedade". Este volume é constituído por oito capítulos cuidadosamente elaborados por professores, pesquisadores, estudantes de graduação e pós-graduação de várias regiões do Brasil. O E-Book explora de maneira profunda tópicos relacionados à Biotecnologia e Saúde, que vão desde a aplicação de tecnologias emergentes na indústria de alimentos até estudos de casos médicos e veterinários. Durante os capítulos, ressalta-se de maneira marcante o papel vital da inovação tecnológica nas Ciências Biológicas e da Saúde. Cada capítulo é uma peça essencial deste mosaico de conhecimento que busca entender e aplicar os princípios da biologia para melhorar nossa qualidade de vida. Este E-Book não apenas enriquecerá sua compreensão, mas também o instigará a explorar as fascinantes áreas das Ciências Biológicas e a forma como elas se entrelaçam com a nossa existência. Esteja convidado a embarcar nessa jornada conosco, a mergulhar e descobrir como as Ciências Biológicas moldam nosso presente e iluminam nosso caminho rumo a um futuro brilhante e sustentável. Clécio Danilo Dias da Silva Daniele Bezerra dos Santos

The operational conditions of β-galactosidase encapsulated in pectin-alginate hydrogel were investigated as well as its catalytic activity in the hydrolysis of lactose from whey and whole milk. β-Galactosidase encapsulated in pectin-alginate hydrogel showed the best results to yields and diffusional effect and pH range. Thermostability showed considerable income gain for half-life (1.7-fold), activation energy of denaturation (31.6-fold), activation of denaturation (217.3-fold at 40 ºC; 266.8-fold at 50 ºC; 345.6-fold at 60 ºC), and entropy of activation of denaturation (−42.0 J mol-1 K-1). Reusability of encapsulated β-galactosidase was observed in 8-cycles for the milk whey lactose hydrolysis (51.9%) and 7-cycles for whole milk (55.6%), and the lactose hydrolysis was observed in 13% in milk whey and 10.3% in whole milk after 10 cycles. These results revealed an industrial application potential of β-galactosidase encapsulated in pectin-alginate hydrogel for lactose hydrolysis of milk whey and whole milk processes.

BACKGROUND: Fructosyltransferase (FTase) enzyme is an important biocatalyst for fructooligossaccaride (FOS) production, but it has low stability in soluble form. The aim of this work was to analyze the stability and kinetics of an fructosyltransferase (FTase E.C.2.4.1.9) from Aspergillus oryzae IPT‐301 immobilized on polyhydroxybutyrate (PHB) and glutaraldehyde‐activated polyhydroxybutyrate (GLU‐PHB).RESULTS: The immobilization yields of the FTase on PHB and GLU‐PHB were about 41% and 55%, respectively. The recovery activities of the FTase immobilized on PHB and GLU‐PHB were about 17% and 11%, respectively. At 30 °C, the FTase immobilized on GLU‐PHB showed a half‐life 2.6 and 1.4 times higher than the soluble FTase and the FTase immobilized on PHB, respectively. The FTase immobilized on PHB and GLU‐PHB retained more than 40% and 55% of their initial activity after six sequential reaction batches, respectively. Both immobilized enzymes showed kinetic behaviors that were fitted by the Hill model, with transfructosylation activity maximum at concentrations between 400 g substrate L⁻¹ and 500 g substrate L⁻¹.CONCLUSION: These results demonstrate that the FTase immobilized on PHB and, especially, on GLU‐PHB, are promising biocatalysts for FOS production in heterogeneous reaction systems. © 2022 Society of Chemical Industry (SCI).

A Plackett-Burman factorial design with 15 experiments was conducted to evaluate the influence of seven factors on lipases production by Fusarium solani. The factors investigated were peptone, tryptone, yeast extract, calcium chloride, potassium phosphate, magnesium sulfate, and copper sulphate. Five fixed variables (cotton oil, pH, temperature, agitation, and time) were maintained and as a response to the enzymatic activity. The concentration of tryptone, calcium chloride, and magnesium sulphate had a significant effect (p < 0.10) on lipase production and was studied consecutively through a complete DCCR (central rotational compound design), to optimize lipase production of the fungi F. solani. After optimization using DCCR, maximum lipolytic activities of 24.84 U/ml were obtained with the use of 10 g.L^-1 tryptone, 3.50 g.L^-1 calcium chloride and 0.50 g.L^-1 magnesium sulfate, 1 g.L^-1 potassium phosphate and 1% soybean oil. The statistical model showed a correlation of 85.67% with the experimental data. The biochemical characterization of lipase showed that the enzyme has a better performance at pH 7 at a temperature of 40 °C, where the statistical model had a correlation of 94.15% with the experimental data. In this way, lipases produced by F. solani have potential for application and use in biodiesel production.

BACKGROUND: Fructooligosaccharides (FOS) are prebiotic sugars used in the production of functional foods. These sugars can be produced by the transfructosylation reaction of sucrose catalyzed by fructosyltransferase enzymes (FTase, E.C.2.4.1.9). The aim of this work was to evaluate the use of cells of Aspergillus oryzae IPT‐301 as biocatalysts for the sucrose transfructosylation reaction in a packed bed reactor (PBR).RESULTS: The morphology of the cells was observed by scanning electron microscopy, and BET specific surface areas were examined by nitrogen physisorption The highest transfructosylation activity (660 U g⁻¹) of the cells was obtained at 25 min of reaction by adopting a reaction temperature of 50 °C, sucrose concentration of 470 g L⁻¹, and volumetric flow of 15 mL min⁻¹. The rise in volumetric flow provided an increase in the transfructosylation activity, but also a faster deactivation of the biocatalyst. Enzyme kinetics was described using the Michaelis–Menten model, with a V_max of 632.8 U g⁻¹ and a k_m of 160.7 g L⁻¹. The cells showed constant enzymatic activity for 12 h of reaction in the PBR operated with discontinuous flow at flow rates of 5 mL min⁻¹ and 11.5 mL min⁻¹, and in the PBR operated with continuous flow at a flow rate of 5 mL min⁻¹. The transition from diffusional to kinetic regimes was observed starting from 11.5 mL min⁻¹.CONCLUSION: The results obtained suggest a high potential of application of the whole A. oryzae IPT‐301 cells for continuous FOS production in PBR. © 2022 Society of Chemical Industry (SCI).

Summary: The use of wild Saccharomyces and non‐Saccharomyces yeasts might result in bread with different and attractive sensory characteristics. This study aimed to evaluate the performance of Saccharomyces and non‐Saccharomyces yeasts as starter culture in dough fermentation to bread making and the physicochemical parameters and aromatic profile of bread. All 26 wild yeasts strains isolated from Brazilian Cerrado fruit and tree bark were osmotolerant, and 19.4% were able to ferment maltose. Candida tropicalis ART101.3 and Saccharomyces cerevisiae SC5952 had the best growth capacity under high concentrations of glucose and maltose. Also, they were resistant to lyophilisation. Kinetic parameters of bioreactor cultivations showed high cell growth and lower generation time with 10 g L⁻¹ maltose. Bread produced with C. tropicalis ART101.3 and the control bread had similar physicochemical properties and acceptance of consumers. Bread with S. cerevisiae SC5951 had a lower specific volume and a different colour than control bread; however, the consumers found no significant difference. More than 70% of the consumers demonstrated purchase intention of bread produced with both wild yeasts. The present study shows the potential of native Cerrado yeasts to be used and exploited in industrial processes and contributes to the diversification of bread starter cultures.

The development of microbial enzymes is of great importance in biochemical process industries. Among the enzymes of industrial importance, cellulolytic and xylanolytic enzymes stand out, which are responsible for the degradation of lignocellulosic material that is usually discarded by the agro-industrial sector. In order to produce the enzymes CMCase, β-glycosidase, xylanase and β-xylosidase, the fungus Fusarium oxysporum was used in liquid fermentation using pre-treated pineapple crown biomass as a carbon source. In order to optimize the production of these enzymes, cultures were carried out for 168 hours, 30 °C at 180 rpm. The biomass underwent a pre-treatment in two stages, the first being submitted to 121 °C for 7 min, while the second was added diluted sulfuric acid to 1%, in the proportion of 1:5 (m/v), at 121 °C for 27 minutes. The partial characterization of the crude extract showed a temperature of 50 ºC as the most satisfactory for the action of these enzymes, and the ideal pH range ranged from 3.5 to 5.5. The enzymatic hydrolysis of pineapple crown, with and without pretreatment, was performed using lyophilized crude extract (CMCase = 9.78 U/mL, β-glucosidase = 10.86 U/mL, xylanase = 27.90 U/mL and β-xylosidase = 0.018 U/ml). An experimental design of the Central Composite Rotational Design was carried out. It was possible to evaluate the effect and influence of pH, biomass concentration and incubation time on the release of glucose in the medium (R2>0.85); pH and biomass (%) had a significant effect on glucose release.