Evaluation of the microencapsulation process of conidia of Trichoderma asperellum by spray drying.

Microencapsulation of microorganisms has been studied to increase product shelf life and stability to enable the application in sustainable agriculture. In this study, the microencapsulation of Trichoderma asperellum conidia by spray drying (SD) was evaluated. The objective was to assess the effect of drying air temperature and wall material (maltodextrin DE20, MD20) concentration on the microencapsulation and to identify the optimum conditions to produce, in high yield, microparticles with low moisture, high conidial viability and conidial survival. Microparticles were characterized in terms of morphology, particle size, and shelf life. A central composite rotatable design (CCRD) was used to evaluate the effect of operating parameters on drying yield (DY), moisture content, conidial viability (CV), and conidial survival (SP). Microencapsulation experiments were carried out under optimum conditions to validate the obtained model. The optimum temperature and MD20/conidia dry weight ratios were 80 °C and 1:4.5, respectively, which afforded a drying yield of 63.85 ± 0.86%, moisture content of 4.92 ± 0.07%, conidial viability of 87.10 ± 1.16%, and conidial survival of 85.78 ± 2.88%. Microencapsulation by spray drying using MD20 as wall material extended the viability of conidia stored at 29 °C compared with the control. The mathematical models accurately predicted all the variables studied, and the association of the microencapsulation technique using DE20 maltodextrin was able to optimize the process and increase the product's shelf life. It was also concluded that high inlet air temperatures negatively affected conidia survival, especially above 100 °C.

Immobilization of ß-galactosidase from Bacillus licheniformis for application in the dairy industry.

The food industry has developed a wide range of products with reduced lactose to allow people with intolerance to consume dairy products. Although ß-galactosidase has extensive applications in the food, pharma, and biotechnology industries, the enzymes are high-cost catalysts, and their use makes the process costly. Immobilization is a viable strategy for enzyme retention inside a reactor, allowing its reuse and application in continuous processes. Here, we studied the immobilization of ß-galactosidase from Bacillus licheniformis in ion exchange resin. A central composite rotational design (CCRD) was proposed to evaluate the immobilization process in relation to three immobilization solution variables: offered enzyme activity, ionic strength, and pH. The conditions that maximized the response were offered enzyme activity of 953 U, 40 mM ionic strength, and pH 4.0. Subsequently, experiments were performed to provide additional stabilization for biocatalyst, using a buffer solution pH 9.0 at 25 °C for 24 h, and crosslinking with different concentrations of glutaraldehyde. The stabilization step drastically impacted the activity of the immobilized enzyme, and the reticulation with different concentrations of glutaraldehyde showed significant influence on the activity of the immobilized enzyme. In spite of substantially affecting the initial activity of the immobilized enzyme, higher reagent concentrations (3.5 g L-1) were effective for maintaining stability related to the number of cycles of the enzyme immobilized. The ß-galactosidase from Bacillus licheniformis immobilized in Duolite A568 is a promising technique to produce reduced or lactose-free dairy products, as it allows reuse of the biocatalyst, decreasing operational costs.Key Points• Immobilization of ß-galactosidase from Bacillus licheniformis in batch reactor• Influence of buffer pH and ionic concentration and offered enzyme activity on immobilization• Influence of glutaraldehyde on operational stability.

High Gravity and Very High Gravity Fermentation of Sugarcane Molasses by Flocculating Saccharomyces cerevisiae: Experimental Investigation and Kinetic Modeling.

Substantial progress has been made in ethanol fermentation technology under high gravity (HG) and very high gravity (VHG), which offer environmental and economic benefits. HG and VHG processes increase the productivity of ethanol, reduce distillation costs, and enable higher yields. The aim of the present study was to evaluate the use of sugarcane molasses as the medium component along with flocculating yeasts for fermentation in a fed-batch process employing this promising technology. We evaluated fed-batch fermentation, HG, and VHG involving a molasses-based medium with high concentrations of reducing sugars (209, 222, and 250 g/L). Fermentation of 222 g/L of total reducing sugars achieved 89.45% efficiency, with a final ethanol concentration of 104.4 g/L, whereas the highest productivity (2.98 g/(L.h)) was achieved with the fermentation of 209 g/L of total reducing sugars. The ethanol concentration achieved with the fermentation of 222 g/L of total reducing sugars was close to the value obtained for P'max (105.35 g/L). The kinetic model provided a good fit to the experimental data regarding the fermentation of 222 g/L. The results revealed that sugarcane molasses and flocculating yeasts can be efficiently used in HG fermentation to reduce the costs of the process and achieve high ethanol titers.

Improvement of ethanol production in fed-batch fermentation using a mixture of sugarcane juice and molasse under very high-gravity conditions.

Ethanol fermentation in very high gravity (VHG) saves energy consumption for ethanol distillation. As the technology offers high ethanol yield and low waste generation and it can be operated at low cost, it could be more efficient at an industrial scale than other ethanol production methods. This work studied ethanol production using a fed-batch bioreactor with a working volume of 1.5 L. The main objective of this research was evaluate the effects of temperature, sugar concentration, and cellular concentration using a Central Composite Design (CCD). Experimental conditions were selected using the surface response technique obtained from the CCD, and the results were validated to test the reproducibility. The following operating conditions were selected: temperature of 27.0 °C, sugar concentration 300.0 g/L, and cell concentration 15.0% (v/v). Under these conditions, after 30 h of fermentation the ethanol concentration, productivity and yield were 135.0 g/L, 4.42 g/(L·h) and 90.0%, respectively. All sugar was completely consumed.

Optimization of the production and characterization of lipase from Candida rugosa and Geotrichum candidum in soybean molasses by submerged fermentation.

This present work describes the production and biochemical characterization of lipase by Candida rugosa and Geotrichum candidum in a culture supplemented with soybean molasses. After optimizing the fermentation times for both microorganisms, the effects of changing the soybean molasses concentration, the fermentative medium pH and the fermentation temperature were evaluated using the Central Composite Planning. When soybean molasses was used at a concentration of 200 g/L at 27 ± 1 °C and pH 3.5, the lipolytic activity measured in the broth was 12.3 U/mL after 12 h for C. rugosa and 11.48 U/mL after 24 h for G. candidum. The molecular masses were 38.3 kDa to G. candidum lipase and 59.7 kDa to C. rugosa lipase, determined by SDS-PAGE. The lipase from both microorganisms exhibited maximal hydrolytic activity at a temperature of 40 °C and were inhibited at pH 10.0. Using different concentration of p-nitrophenylbutyrate (p-NPB), the kinetic parameters were calculated, as follows: the Km of lipase from G. candidum was 465.44 µM and the Vmax 0.384 µmol/min; the Km and Vmax of lipase from C. rugosa were 129.21 µM and 0.034 µmol/min, respectively. Lipases activity were increased by metallic ions Mg(2+) and Na(+) and inhibited by metallic ion Cu(3+).

Alcoholic fermentation with flocculant Saccharomyces cerevisiae in fed-batch process.

Studies have been conducted on selecting yeast strains for use in fermentation for ethanol production to improve the performance of industrial plants and decrease production costs. In this paper, we study alcoholic fermentation in a fed-batch process using a Saccharomyces cerevisiae yeast strain with flocculant characteristics. Central composite design (CCD) was used to determine the optimal combination of the variables involved, with the sucrose concentration of 170 g/L, a cellular concentration in the inoculum of 40% (v/v), and a filling time of 6 h, which resulted in a 92.20% yield relative to the theoretical maximum yield, a productivity of 6.01 g/L h and a residual sucrose concentration of 44.33 g/L. With some changes in the process such as recirculation of medium during the fermentation process and increase in cellular concentration in the inoculum after use of the CCD was possible to reduce the residual sucrose concentration to 2.8 g/L in 9 h of fermentation and increase yield and productivity for 92.75% and 9.26 g/L h, respectively. A model was developed to describe the inhibition of alcoholic fermentation kinetics by the substrate and the product. The maximum specific growth rate was 0.103 h(-1), with K(I) and K(s) values of 109.86 and 30.24 g/L, respectively. The experimental results from the fed-batch reactor show a good fit with the proposed model, resulting in a maximum growth rate of 0.080 h(-1).

Evaluation of hexavalent chromium removal in a continuous biological filter with the use of central composite design (CCD).

Hexavalent chromium is frequently found in industrial effluents as a result of the industrial applications of this compound and its anti-corrosive features. However, hexavalent chromium is extremely toxic, and its discharge in water is regulated, with a maximum limit of 0.1 mg/L in accordance with legislation established by CONAMA-Brazil (no. 397, April 3, 2008). To achieve lower discharge values, it is necessary to reduce from Cr(VI) to Cr(III), which is less toxic, and an economic alternative involves biological removal of this compound. Residence time distributions (RTDs) were measured to evaluate the behavior of actual biofilter operation conditions in a biofilter flow. The medium residence time distributions used were 8 and 24 h (recommended by the legislation). To optimize this process, a central composite design was used, considering the initial chromium concentration and pH as the independent variables and the removal of hexavalent chromium as the response. The boundary curves and surface response showed optimal behavior at 3.94 mg/L [Cr(0)] and a pH of 6.2. The removal process of hexavalent chromium is mathematically described by the Michaelis-Menten kinetic model. This model appropriately represents the variation of chromium concentration along the bioreactor.

Microbial rennet produced by Mucor miehei in solid-state and submerged fermentation

A produção de renina microbiana por Mucor miehei foi estudada através de Fermentação Submersa e em Estado Sólido. O objetivo deste trabalho foi verificar o efeito de diferentes fontes de carbono e nitrogênio, utilizando Fermentação Submersa e da adição de caseína utilizando Fermentação em Estado Sólido na produção da renina microbiana. Os picos de biomassa foram de 6,7; 8,1 e 8 g/L e atividade enzimática de 1.066; 857 e 480 Unidades Soxhlet (U.S.) para as concentrações de glicose: 18, 25 e 35 g/L respectivamente. Em frascos aletados, os picos de biomassa foram de 6,7; 8,3 e 10 g/L e atividade enzimática de 648, 279 e 300 U.S., para a mesma concentração de glicose. Quando se utilizou Proflo (Farinha de semente de algodão, Traders ®) e Água de Maceração de Milho os picos de atividade enzimática foram de 667 e 923 U.S., respectivamente. Nos experimentos utilizando Fermentação em Estado Sólido com a adição de HCl 0,2 N a máxima atividade enzimática foi de 414 U.S. e, quando utilizou-se caseína (1 e 2 gramas), verificou-se valores mais altos de atividade: 966 e 1117 U.S respectivamente. Os resultados sugerem que o aumento na concentração de glicose afeta a síntese da enzima e que a caseína é um importante fator na indução neste processo. Fermentação em Estado Sólido pode ser considerada uma boa opção para a produção de renina.