Microbiological contamination profile in soft drinks.

Soft drinks are food matrices propitious to the growth of acidophilic bacteria, yeasts, and filamentous fungi due to their pH, water activity, and the presence of nutrients. Off-flavor, clouding, and package stuffing are the only parameters producers have to detect spoilage when it is often too late for the brand's reputation. In this work, microbiological analyses were performed on non-alcoholic beverages of Brazilian and Bolivian brands. As a result, Gram-positive, Gram-negative, yeast, and filamentous fungi were isolated. Zygosaccharomyces bisporus yeast was isolated from different flavored stuffed products, and Gluconacetobacter liquefaciens and Brevibacillus agri were isolated from packages without visible signs of deterioration. These microorganisms were identified by MALDI-TOF. For products with visible growth of filamentous fungi, microscopic identification keys identified Aspergillus flavus, Penicillium citrinum, Paecilomyces niveus, and Paecilomyces variotii. These work's findings reflect a failure to sanitize raw materials since the isolates' primary origin is the soil and the water, pointing to the lack of process control in soft drinks.

In situ product recovery techniques aiming to obtain biotechnological products: A glance to current knowledge.

Biotechnology and bioengineering techniques have been widely used in the production of biofuels, chemicals, pharmaceuticals, and food additives, being considered a "green" form of production because they use renewable and nonpolluting energy sources. On the other hand, in the traditional processes of production, the target product obtained by biotechnological routes must undergo several stages of purification, which makes these processes more expensive. In the past few years, some works have focused on processes that integrate fermentation to the recovery and purification steps necessary to obtain the final product required. This type of process is called in situ product recovery or extractive fermentation. However, there are some differences in the concepts of the techniques used in these bioprocesses. In this way, this review sought to compile relevant content on considerations and procedures that are being used in this field, such as evaporation, liquid-liquid extraction, permeation, and adsorption techniques. Also, the objective of this review was to approach the different configurations in the recent literature of the processes employed and the main bioproducts obtained, which can be used in the food, pharmaceutical, chemical, and/or fuel additives industry. We intended to elucidate concepts of these techniques, considered very recent, but which emerge as a promising alternative for the integration of bioprocesses.

Enzyme-assisted extraction of carotenoids and phenolic compounds from sunflower wastes using green solvents.

The aim of this work is to develop an optimized enzymatic assisted extraction methodology to extract carotenoids and phenolic compounds from sunflower wastes (petals and florets) using natural hydrophobic green solvents. Several natural green hydrophobic solvents were used as well as natural hydrophobic eutectic solvents composed of d,l-menthol and different acids, with different hydrophobicity. The multi-enzyme complex Viscozyme® was used to disrupt the cell wall of petals and disc florets. The extracted carotenoids content into the hydrophobic phase was quantified using UV-Vis spectrophotometry and the carotenoids profile was studied using high-performance liquid and thin layer chromatography. The amount of total sugars in the aqueous phase was also analyzed using the dinitrosalicylic acid (DNS) method to infer about the enzymatic action in cell wall. Phenolic compounds also in the aqueous phase were analyzed by Folin Denis method. The eutectic solvent d,l-menthol:d,l-lactic acid (M:HLac) (1:2) was the best solvent for extraction of carotenoids from sunflower wastes, with 147 ppm of carotenoids extracted, in comparison to 115 ppm obtained with the standard solvent, n-hexane. In what concerns phenolic compounds, M:HLac was again better than the standard solvent. The use of the multi-enzyme complex Viscozyme® had different responses, depending on the solvent tested. For the green solvent M:HLac, the enzyme improved the carotenoids extraction, achieving 335 ppm of carotenoids in the extract. The role of enzyme, solvent, water and sunflower quantity in the carotenoid extraction was evaluated and optimized through a central composite rotatable design (CCRD), using the M:HLac as solvent. According to the analysis of CCRD, the most efficient extractions were carried out using more solvent and less raw material, whose best result reached 1449 mg carotenoids/100 g biomass ppm of carotenoids. This work emphasizes the possibility of developing more sustainable enzyme-assisted separation processes, through the substitution of toxic solvents with natural, environmentally friendly, solvents.

A novel osmotic pressure strategy to improve erythritol production by Yarrowia lipolytica from glycerol.

Polyethylene glycol (PEG) is polymer that was used to replace NaCl (reference media) as an osmotic stress agent for the synthesis of erythritol by the osmophilic yeast Yarrowia lipolytica. Two strains, the wild-type strain IMUFRJ 50682 and the lab strain W29, were grown in the presence of PEG of different molecular weights. For strain IMUFRJ 50682, the erythritol titer was increased by 40% in the presence of PEG2000 as compared to the reference media (with NaCl). A similar increase was also observed for strain W29, except that it occurred in the presence of PEG6000. Moreover, in those experimental conditions neither strain produced mannitol, in contrast to the control medium. These results highlight that PEG could be used to increase erythritol productivity and to simultaneously inhibit mannitol synthesis, representing a good substitute for NaCl as an osmotic stress agent.

Green coconut fiber: a novel carrier for the immobilization of commercial laccase by covalent attachment for textile dyes decolourization.

Commercial laccase formulation was immobilized on modified green coconut fiber silanized with 3-glycidoxypropyltrimethoxysilane, aiming to achieve a cheap and effective biocatalyst. Two different strategies were followed: one point (pH 7.0) and multipoint (pH 10.0) covalent attachment. The influence of immobilization time on enzymatic activity and the final reduction with sodium borohydride were evaluated. The highest activities were achieved after 2 h of contact time in all situations. Commercial laccase immobilized at pH 7.0 was found to have higher activity and higher affinity to the substrate. However, the immobilization by multipoint covalent attachment improved the biocatalyst thermal stability at 50 °C, when compared to soluble enzyme and to the immobilized enzyme at pH 7.0. The Schiff's bases reduction by sodium borohydride, in spite of causing a decrease in enzyme activity, showed to contribute to the increase of operational stability through bonds stabilization. Finally, these immobilized enzymes showed high efficiency in the continuous decolourization of reactive textile dyes. In the first cycle, the decolourization is mainly due to dyes adsorption on the support. However, when working in successive cycles, the adsorption capacity of the support decreases (saturation) and the enzymatic action increases, indicating the applicability of this biocatalyst for textile wastewater treatment.

Image analysis application for the study of activated sludge floc size during the treatment of synthetic and real fishery wastewaters.

BACKGROUND, AIM, AND SCOPE: Fishery wastewater treatment can be compromised due to seasonal production. The use of sequencing batch reactors is not completely successful, despite flexibility being one of the principal advantages. Most research on activated sludge is performed using synthetic wastewater to ensure a stable and constant feed. The current work compared biomass morphology and settling ability using image analysis of synthetic and real fishery wastewaters, with and without NaCl addition. RESULTS: The final effluent presented higher turbidity for both wastewaters after NaCl addition, and lower SVI values. For synthetic wastewater, NaCl addition led to the total aggregates' area (TA) increase from 1.46 to 2.09 mm(2)/µL, alongside the growth of intermediate aggregates into larger aggregates. The addition of NaCl to the fishery wastewater led to a decrease of the TA from 4.43 to 1.72 mm(2)/µL. The biomass composition decreased in larger and intermediate structures, opposite to the smaller aggregates' area percentage increase. DISCUSSION: NaCl addition to synthetic wastewater incited flocculation increasing sludge settling ability. A slight aggregate disruption was responsible for a turbidity increase. A strong deflocculation was identified in fishery wastewater with NaCl from the decrease of intermediate and large aggregates. This contrasted with pinpoint flocs release, which increased the turbidity levels. CONCLUSIONS, RECOMMENDATIONS, AND PERSPECTIVES: It could be established that synthetic wastewater biomass flocculation and fishery wastewater biomass deflocculation, observed during 0.5% NaCl experiments, were related to sludge settling and effluent turbidity changes. Furthermore, the biomass changes obtained with synthetic wastewater cannot be extrapolated to fishery wastewater.

Biosurfactants from yeasts: characteristics, production and application.

Biosurfactants are surface-active compounds from biological sources, usually extracellular, produced by bacteria, yeast or fungi. Research on biological surfactant production has grown significantly due to the advantages they present over synthetic compounds such as biodegradability, low toxicity, diversity of applications and functionality under extreme conditions. Although the majority of microbial surfactants have been reported in bacteria, the pathogenic nature of some producers restricts the wide application of these compounds. A growing number of aspects related to the production of biosurfactants from yeasts have been the topic of research during the last decade. Given the industrial importance of yeasts and their potential to biosurfactant production, the goal of this chapter is to review the biosurfactants identified up to present, focusing the relevant parameters that influence biosurfactant production by yeasts and its characteristics, revealing the potential of application of such compounds in the industrial field and presenting some directions for the future development of this area, taking into account the production costs.

Optimization of oxygen mass transfer in a multiphase bioreactor with perfluorodecalin as a second liquid phase.

Oxygenation is an important parameter involved in the design and operation of mixing-sparging bioreactors and it can be analyzed by means of the oxygen mass transfer coefficient (k(L)a). The operational conditions of a stirred, submerged aerated 2-L bioreactor have been optimized by studying the influence of a second liquid phase with higher oxygen affinity (perfluorodecalin or olive oil) in the k(L)a. Using k(L)a measurements, the influence of the following parameters on the oxygen transfer rate was evaluated: the volume of working medium, the type of impellers and their position, the organic phase concentration, the aqueous phase composition, and the concentration of inactive biomass. This study shows that the best experimental conditions were achieved with a perfluorodecalin volume fraction of 0.20, mixing using two Rushton turbines with six vertical blades and in the presence of YPD medium as the aqueous phase, with a k(L)a value of 64.6 h(-1). The addition of 20% of perfluorodecalin in these conditions provided a k(L)a enhancement of 25% when pure water was the aqueous phase and a 230% enhancement when YPD medium was used in comparison to their respective controls (no perfluorodecalin). Furthermore it is shown that the presence of olive oil as a second liquid phase is not beneficial to the oxygen transfer rate enhancement, leading to a decrease in the k(L)a values for all the concentrations studied. It was also observed that the magnitude of the enhancement of the k(L)a values by perfluorodecalin depends on the biomass concentration present.

Electrical stimulation of saccharomyces cerevisiae cultures

Modulação do potencial de membrana celular endógeno por um campo elétrico externo influencia a estrutura e função dos compartimentos da membrana, de suas proteínas e da bi-camada lipídica. Neste trabalho, os efeitos da aplicação de potencial no crescimento de Saccharomyces cerevisiae foram caracterizados por experimentos simples, mas conclusivos. O perfil temporal de crescimento celular e a divisão celular foram investigados como respostas macroscópicas ao estímulo elétrico. Experimentos controle foram conduzidos em condições idênticas, exceto pela ausência de potencial aplicado. Através de análise comparativa, verificou-se que o estímulo elétrico alterou o ciclo celular como foi possível observar através da medida da dispersão de tamanho celular de cada população, sugerindo um possível sincronismo na divisão celular. Análise do espectro de potência foi empregada para sustentar o aumento no sincronismo, e uma modelagem matemática foi conduzida para determinar mudanças na cinética de crescimento celular. Parâmetros cinéticos do modelo tipo Monod para crescimento foram determinados por regressão não-linear. A constante de afinidade (a saber, KS) apresentou uma dependência com o potencial aplicado, sugerindo mudanças no transporte através da membrana celular. Verificou-se, também, que o estresse promovido eletroquimicamente inibiu o crescimento e induziu mudanças na viabilidade celular.

Partial purification of a polygalacturonase produced by solid-state cultures of Aspergillus niger 3T5B

A poligalacturonase produzida em cultura semi sólida de Aspergillus niger 3T5B8 foi purificada usando fracionamento por adiçäo de sal (salting out), diálise e cromatografia de gel-filtraçäo. A adiçäo de sulfato de amônio em diferentes níveis de saturaçäo apresentaram um compromisso entre purificaçäo e recuperaçäo da atividade enzimática. A cromatografia por gel-filtraç o forneceu bons resultados quando azida de sódio foi utilizada nos tampöes de eluiçäo, diminuindo as perdas na atividade enzimática em 14 por cento. O peso molecular determinado para esta poligalacturonase foi de 34700 daltons