New frontiers of soil fungal microbiome and its application for biotechnology in agriculture.

The fungi-based technology provided encouraging scenarios in the transition from a conventionally based economic system to the potential security of sources closely associated with the agricultural sphere such as the agriculture. In recent years, the intensification of fungi-based processes has generated significant gains, additionally to the production of materials with significant benefits and strong environmental importance. Furthermore, the growing concern for human health, especially in the agriculture scenario, has fostered the investigation of organisms with high biological and beneficial potential for use in agricultural systems. Accordingly, this study offered a comprehensive review of the diversity of the soil fungal microbiome and its main applications in a biotechnological approach aimed at agriculture and food chain-related areas. Moreover, the spectrum of opportunities and the extensive optimization platform for obtaining fungi compounds and metabolites are discussed. Finally, future perspectives regarding the insurgency of innovations and challenges on the broad rise of visionary solutions applied to the biotechnology context are provided.

Extraction and characterization of polysaccharide-enriched fractions from Phoma dimorpha mycelial biomass.

Ultrasound-assisted extraction (UAE) and pressurized hot water extraction (PHWE) were tested as advanced clean methods to obtain polysaccharides from Phoma dimorpha mycelial biomass. These methods were compared to conventional extraction (hot water extraction, HWE) in terms of polysaccharides-enriched fractions (PEF) yield. A central composite rotational design was performed for each extraction method to investigate the influence of independent variables on the yield and to help the selection of the condition with the highest yield using water as an extraction solvent. The best extraction condition of PEF yielded 12.02 wt% and was achieved when using UAE with direct sonication for 30 min under the intensity of 75.11 W/cm2 and pulse factor of 0.57. In the kinetic profiles, the highest yield (15.28 wt%) was obtained at 50 °C under an ultrasound intensity of 75.11 W/cm2 and a pulse factor of 0.93. Structural analysis of extracted polysaccharide was performed using Fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and thermal property. The water solubility index, water holding capacity, and emulsification index of PEF were 31.3 ± 1.5%, 138.1 ± 3.2%, and 62.9 ± 2.3%, respectively. The submerged fermentation demonstrates the huge potential of Phoma dimorpha to produce polysaccharides with bioemulsifying properties as a biotechnologically cleaner alternative if compared to commercial petroleum-derived compounds. Furthermore, UAE and PHWE are green technologies, which can be operated at an industrial scale for PEF extraction.

Ultrasound as an alternative method to increase the extraction yield from chicken mecanically separated meatresidue collagen.

The residue from chicken mechanically separated meat (MSM) is a potential source for the extraction of collagen. However, this process requires the removal of many covalent crosslinks, which makes it quite complex. Ultrasound has been successfully used to extract collagen; it reduces the process time and increases the yield. However, information regarding the effects of this treatment on the structural and functional properties of proteins is still very limited. Therefore, the aims of the present study were to obtain collagen from chicken MSM residue and to test the effects of pre-treatment with ultrasonic probe and enzymatic extraction with pepsin in its yield, as well as to evaluate the properties of extracted collagen using gel electrophoresis, Fourier-transform infrared spectroscopy, solubility, and differential scanning calorimetry. Both the ultrasound and the enzymatic extraction had a positive effect on the extraction yield of collagen from chicken MSM residue without affecting its integrity. Using ultrasound led to an increase of up to 40% in yield when compared to treatments without ultrasound application. Five extraction treatments were considered. The extracted collagen exhibited high thermal stability (43.9-47.0 °C) and mainly type I structure. The use of ultrasound as pre-treatment, together with enzymatic extraction with pepsin, were effective in increasing the extraction yield of collagen from chicken MSM residue, as well as preserving the triple helical structure of the native collagen.

An overview of fungal biopolymers: bioemulsifiers and biosurfactants compounds production.

Fungal biopolymers have gained considerable attention from the scientific community for various applications due to their biological and physicochemical properties. The wide applications in several areas, especially in the food industry as a bioemulsifier and in the agricultural area as a biosurfactant, have expanded the knowledge on the production of fungal biopolymers to keep up with developments on this subject area. Recent scientific studies have disclosed novel routes, optimized parameters, increased yields, and other related approaches in order to produce and apply fungal bioemulsifiers and biosurfactants. However, there is a need to gather important information in order to provide a way forward. Therefore, this review presents an overview of properties, applications, and perspectives for encouraging further projects and investments in the near future by most categories of investors. The selection of culture media, the definition of cultivation parameters, extraction, recovery, and purification are the initial steps to indicate the conditions for scale-up. Indeed, scale-up is still one of the challenges in this biotechnological field, which could be solved by expanding the tests and operational productions in both pilot and industrial plants.

Improving the soluble lipase-catalyzed biodiesel production through a two-step hydroesterification reaction system.

The application of lipases in liquid formulation instead of immobilized forms in the enzymatic biodiesel synthesis can make the process cost-efficient, more competitive, and sustainable. However, despite the benefits, the long reaction times required to achieve satisfactory yields is still a drawback of this biotechnological process. In this sense, employing the novel low-cost soluble NS40116 lipase, this paper proposes an innovative two-step hydroesterification reaction (TSHR) system as a technique of improving the reaction rate of an enzymatic biodiesel production. With the employment of two central composite statistical design to optimize the parameters of each of the reactions involved, the influence of the parameters "water concentration added to the reaction," "methanol-to-oil molar ratio," and "lipase load" on the process yield, besides the acid value of the samples, was investigated. After only 8 h of reaction, the highest fatty acid methyl ester yield reached was 97.1% with an acid value of 4.62 mg KOH g-1 utilizing a total of 8 wt% water, methanol-to-oil molar ratio of 6.3:1, and 0.70 wt% of lipase. Furthermore, the statistical models for both reactions indicated to be significant with 95% of reliability. Considering that the papers published using soluble lipases in a one-step batch process normally reach similar yields to those obtained in this research after 16 h to 24 h of reaction, the proposed system demonstrated to be a promising option of process configuration for the enzymatic production of biodiesel.

Production of cutinase by solid-state fermentation and its use as adjuvant in bioherbicide formulation.

In the present study, it was presented a strategy to maximize the cutinase production by solid-state fermentation from different microorganisms and substrates. The best results were observed using Fusarium verticillioides, rice bran being the main substrate. Maximum yield of cutinase obtained by the strain was 16.22 U/g. For concentration, ethanol precipitation was used, and the purification factor was 2.4. The optimum temperature and pH for enzyme activity were 35 °C and 6.5, respectively. The enzyme was stable at a wide range of temperature and at all pH values tested. The concentrated cutinase was used as an adjuvant in a formulation containing cutinase + bioherbicide. The use of enzyme increased the efficiency of bioherbicide, since cutinase was responsible to remove/degrade the cutin that recovery the weed leaves and difficult the bioherbicide absorption. Cutinase showed to be a promising product to be used in formulation of bioherbicides.

Cellulolytic enzyme production from agricultural residues for biofuel purpose on circular economy approach.

This study evaluated the production of cellulolytic enzymes from different agricultural residues. The crude enzyme extract produced was characterized and applied for saccharification of some agricultural residues. Maximum cellulolytic activities were obtained using soybean hulls. All enzymatic activities were highly stable at 40 °C at a pH range of 4.5-5.5. For stability at low temperatures, the enzyme extract was stored at freezing temperature and cooling for about 290 days without major loss of activity. The Km values found for total cellulase (FPase), endoglucanase (CMCase), and xylanase were 19.73 mg ml-1, 0.65 mg ml-1, and 22.64 mg ml-1, respectively, and Vmax values were 0.82 mol min-1 mg-1, 0.62 mol min-1 mg-1, and 104.17 mol min-1 mg-1 to cellulose, carboxymethyl cellulose, and xylan, respectively. In the saccharification tests, the total amount of total reducing sugars (TRS) released from 1 g of soybean hulls catalyzed by the enzymes present in the crude enzyme extract was 0.16 g g-1 dry substrate.

Soluble lipase-catalyzed synthesis of methyl esters using a blend of edible and nonedible raw materials.

This work investigates the use of blends of edible and nonedible raw materials as an alternative feedstock to fatty acid methyl esters (FAME) production through enzymatic catalysis. As biocatalyst, liquid lipase from Thermomyces lanuginosus (Callera™ Trans L), was used. Under reaction conditions of 35 °C, methanol to feedstock molar ratio of 4.5:1 and 1.45% of catalyst load, the best process performance was reached using 9% of water concentration in the medium-yield of 79.9% after 480 min of reaction. In terms of use of tallow mixed with soybean oil, the best yield was obtained when 100% of tallow was used in the process-84.6% after 480 min of reaction-behavior that was associated with the degree of unsaturation of the feedstock, something by that time, not addressed in papers of the area. The results show that tallow can be used as an alternative to FAME production, catalyzed by soluble lipase.

Modeling the microbial growth and temperature profile in a fixed-bed bioreactor.

Aiming to scale up and apply control and optimization strategies, currently is required the development of accurate plant models to forecast the process nonlinear dynamics. In this work, a mathematical model to predict the growth of the Kluyveromyces marxianus and temperature profile in a fixed-bed bioreactor for solid-state fermentation using sugarcane bagasse as substrate was built up. A parameter estimation technique was performed to fit the mathematical model to the experimental data. The estimated parameters and the model fitness were evaluated with statistical analyses. The results have shown the estimated parameters significance, with 95 % confidence intervals, and the good quality of process model to reproduce the experimental data.

Carbon nanotubes as supports for inulinase immobilization.

The commercial inulinase obtained from Aspergillus niger was non-covalently immobilized on multiwalled carbon nanotubes (MWNT-COOH). The immobilization conditions for the carbon nanotubes were defined by the central composite rotational design (CCRD). The effects of enzyme concentration (0.8%-1.7% v/v) and adsorbent:adsorbate ratio (1:460-1:175) on the enzyme immobilization were studied. The adsorbent:adsorbate ratio variable has positive effect and the enzyme concentration has a negative effect on the inulinase immobilization (U/g) response at the 90% significance level. These results show that the lower the enzyme concentration and the higher the adsorbent:adsorbate ratio, better is the immobilization. According to the results, it is possible to observe that the carbon nanotubes present an effective inulinase adsorption. Fast adsorption in about six minutes and a loading capacity of 51,047 U/g support using a 1.3% (v/v) inulinase concentration and a 1:460 adsorbent:adsorbate ratio was observed. The effects of temperature on the immobilized enzyme activity were evaluated, showing better activity at 50 °C. The immobilized enzyme maintained 100% of its activity during five weeks at room temperature. The immobilization strategy with MWNT-COOH was defined by the experimental design, showing that inulinase immobilization is a promising biotechnological application of carbon nanotubes.

Submerged cultivation of Nigrospora sp. in batch and fed-batch modes for microbial oil production.

Microbial lipids are a valuable source of potential biofuels and essential polyunsaturated fatty acids. The optimization of the fermentation conditions is a strategy that affects the total lipid concentration. The genus Nigrospora sp. has been the target of investigations based on its potential bioherbicidal action. Therefore, this study developed a strategy to maximize the biomass concentration and lipid accumulation by Nigrospora sp. in submerged fermentation. Different media compositions and process variables were investigated in shaken flasks and bioreactor in batch and fed-batch modes. Maximum biomass concentration and lipid accumulations were 40.17 g/L and 21.32 wt% in the bioreactor, which was 2.1 and 5.4 times higher than the same condition in shaken flasks, respectively. This study presents relevant information to the production of fungal lipids since few investigations are exploring the fed-batch strategy to increase the yield of fungi lipids, as well as few studies investigating Nigrospora sp. to produce lipids.

Effect of magnetic field on the ultrafiltration of bovine serum albumin.

This work evaluates the effects of a static magnetic field on the permeation of bovine serum albumin (BSA) in a tangential ultrafiltration membrane module. Experimental tests were carried out at different pHs using a poly(sulfone) membrane with molecular weight cut off of 60 kDa under the influence of a 0.4 T neodymium-iron-boron magnetic field. Results showed an increase in the permeate flux of water after the cleaning procedures of the new and reused membranes in the presence of the magnetic field. The elusive mechanism of magnetic memory is also shown to take place for the water fluxes fully recovered after the cleaning procedures when the magnetic field was applied to the system before the permeation. When the magnetic field was applied during permeation, the water fluxes presented lower percent of recuperation after the cleaning procedures, thus suggesting that the BSA solution may have somewhat been influenced by magnetic memory.

Use of papaya seeds as a biosorbent of methylene blue from aqueous solution.

In this study papaya seeds were used to remove methylene blue dye from aqueous solution. Papaya seeds were characterized as possessing a macro/mesoporous texture and large pore size. Studies were carried out in batches to evaluate the effect of contact time and pH (2-12) on the removal of dye. It was observed that the adsorption of dye was better in the basic region (pH 12). The equilibrium data were analyzed using Langmuir, Freundlich, Dubinin-Raduschkevich, Tempkin, Jovanovich, Redlich-Peterson, Sips, Toth and Radke-Prausnitz isotherms. The equilibrium data were best described by the Langmuir isotherm with a maximum adsorption capacity of 637.29 mg g(-1). Adsorption kinetic data were fitted using the pseudo-first-order and pseudo-second-order model. The adsorption kinetic is very fast and was best described by the pseudo-second-order model.

Activated carbon prepared from yerba mate used as a novel adsorbent for removal of tannery dye from aqueous solution.

Activated carbon prepared from yerba mate (Ilex paraguariensis) was used as adsorbent for the removal of tannery dye from aqueous solution. The activated carbon was characterized, and it showed a mesoporous texture, with surface area of 537.4 m2 g(-1). The initial dye concentration, contact time and pH influenced the adsorption capacity. The equilibrium data were in good agreement with both Langmuir and Freundlich isotherms. The adsorption kinetics of the tannery dye on activated carbon prepared from yerba mate followed a pseudo-second-order model. The adsorption process was found to be controlled by both external mass-transfer and intraparticle diffusion, but the external diffusion was the dominating process. This work highlights the potential application of activated carbon produced from yerba mate in the field of adsorption.

Immobilization of inulinase from Kluyveromyces marxianus NRRL Y-7571 using modified sodium alginate beads.

An experimental design was carried out to evaluate the effect of the concentrations of sodium alginate, glutaraldehyde and activated coal on the immobilization of inulinase from Kluyveromyces marxianus NRRL Y-7571. The experimental condition of 20 g/L of sodium alginate, 50 mL/L of glutaraldehyde and 30 g/L of activated coal led to the highest specific activity (2,063.5 U/mg of protein), corresponding to an enhancement of about 26 times compared to the activity of the free enzyme (79.1 U/mg of protein). The effect of pH and temperature on the immobilized enzyme activity was also evaluated, showing optimal activities at pH of 5.5 and 55 °C. The study of storage of immobilized inulinase in different temperatures showed that the extract kept its initial activity after 43 days of storage at 40 and 50 °C and after 138 days of storage either at 4 or 25 °C.

Characterization of a commercial cellulase for hydrolysis of agroindustrial substrates.

This work is focused on the characterization of a commercial cellulase in terms of optimum pH and temperature, stability to pH and temperature and affinity of this enzyme to several substrates, determining the Michaelis-Menten parameters. Maximum activity of cellulase was obtained for the temperature range from 40 to 50 °C and pH from 5.2 to 5.5. Enzyme activity decreased only 15% after 150 h of reaction at temperatures between 30 and 50 °C. No loss of activity was observed at pH 5.0 and 5.5. The cellulase showed satisfactory results in the hydrolysis of agroindustrial substrates, since similar activity was verified on filter paper and other agroindustrial substrates.

Evaluation of ultrasound waves for the production of chitinase and ß-1,3 glucanase by Trichoderma harzianum through SSF.

Cell wall degrading enzymes (chitinase and ß-1,3-glucanase) were produced by solid-state fermentation (SSF) using the fungus Trichoderma harzianum and different agro-industrial products, mainly residues. The influence of temperature (25-35 °C), initial moisture content (50-90% w/w), nutrient solution (1-2% v/w), and yeast extract (1-2% w/w) on enzyme activity was evaluated. The application of ultrasound during fermentation for different times (0-6 h/day) was also studied. White rice was the substrate that showed the highest chitinase and ß-1,3-glucanase activities, which were 31.31 U/g for chitinase and 23.83 U/g for ß-1,3-glucanase after 10 days of fermentation. Application of ultrasound waves during fermentation positively affected (p < 0.05) the enzyme activities. The best results for chitinase (51.88 U/g) and ß-1,3-glucanase (39.22 U/g) were obtained with a 50% (w/w) moisture content and 4 h/day ultrasound application for 10 days of fermentation. Increases of 3.6-fold (from 14.37 to 51.88 U/g) and 3.8-fold (from 10.22 to 39.22 U/g) in activities for chitinase and ß-1,3-glucanase, respectively, compared to non-sonicated fermentation, were obtained. Ultrasound technique associated with the SSF process was a promising alternative to increase the production activity of cell wall degrading enzymes: chitinase and ß-1,3-glucanase.

Weed control by metabolites produced from Diaporthe schini.

Weed control is a critical factor to ensure productivity and quality for food production. Chemical control is the main method used worldwide, but the demand for healthier food and the farmers' health and environment concerns have led to an increase in the search for alternative control methods. In this way, the use of biomolecules produced by microorganisms that present phytotoxic activity against weeds, such as exopolysaccharides, is attracting attention. For this purpose, this work compared two techniques (solid-state and submerged fermentation) for bioherbicide production by Diaporthe schini. Physicochemical characterization of both fermented broth and evaluation of bioherbicidal effect in post-emergence of Amaranthus viridis, Bidens pilosa, Echinocloa crusgalli, and Lollium multiflorum were performed. Fungal broth obtained by submerged fermentation presented better physicochemical characteristics in terms of viscosity, density, and surface tension. Overall, it was more effective than the broth obtained by solid-state fermentation for weed control because it presented an average inhibition of 40% of weed growth and 45% lower surface tension if compared to the control test. Also, reductions of 1.4-4.2 times of root dry mass, 2.9-5.8 times of shoot dry mass and 1.2-3.9 times of weeds heights, if compared to the control test, were achieved.

Powder containing biomolecules from Diaporthe schini for weed control.

This study describes the use of spray drying technology to obtain a powder containing biomolecules with herbicidal activity produced by submerged fermentation using Diaporthe schini. The efficiency of the bioherbicide was tested for the post-emergence control of Bidens pilosa L., Amaranthus viridis L., Echinochloa crusgalli (L.) Beauv., and Lolium multiflorum Lam. In the first step, different additives were used and lactose was the most suitable one because it resulted in high herbicidal activity and weed suppression. In the second step, process variables were investigated, including inlet air temperature, drying air flow rate, and feed flow rate. The highest herbicidal activity was obtained with an inlet air temperature of 100°C, and air and feed flow rates of 1.4 m3/min and 0.22 L/h, respectively. Maximum herbicidal activities were 38, 45, 21 and 18%, while weed heights reduction were 69.0, 74.3, 20.4 and 24.8% for B. pilosa, A. viridis, E. crusgalli and L. multiflorum, respectively. The bioherbicide was effective to suppress weed growth and spray drying is a promising technology for the production of solid formulations of bioherbicides.

Hybrid modeling of xanthan gum bioproduction in batch bioreactor.

This work is focused on hybrid modeling of xanthan gum bioproduction process by Xanthomonas campestris pv. mangiferaeindicae. Experiments were carried out to evaluate the effects of stirred speed and superficial gas velocity on the kinetics of cell growth, lactose consumption and xanthan gum production in a batch bioreactor using cheese whey as substrate. A hybrid model was employed to simulate the bio-process making use of an artificial neural network (ANN) as a kinetic parameter estimator for the phenomenological model. The hybrid modeling of the process provided a satisfactory fitting quality of the experimental data, since this approach makes possible the incorporation of the effects of operational variables on model parameters. The applicability of the validated model was investigated, using the model as a process simulator to evaluate the effects of initial cell and lactose concentration in the xanthan gum production.